Treatment of colorectal cancer

Abstract

The present disclosure relates to methods of treating KRAS G12C-mutated colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

Description

WSGR Ref. 47134-784.601TREATMENT OF COLORECTAL CANCERCROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 778,753 filed March 27, 2025, and U.S. Provisional Application No. 63 / 935,626 filed December 10, 2025, which are hereby incorporated by reference in their entirety.BACKGROUND OF THE INVENTION

[0002] Kirsten rat sarcoma virus oncogene homolog (KRAS) is a commonly mutated oncogene in human cancer. In colorectal cancers, KRAS mutations are present in more than 50% of cases, and the KRAS glycine-to-cysteine mutation at codon 12 (KRAS G12C) occurs in up to 4% of patients. This mutation is associated with short responses to standard chemotherapy and worse overall survival compared to non-G12C mutations. Recently, several KRAS G12C inhibitors, including adagrasib and sotorasib, have been approved for use in the treatment of subjects having KRAS G12C-mutated colorectal cancer. Despite promising efficacy in the treatment of subjects having KRAS G12C-mutated colorectal cancer, the use of KRAS G12C inhibitors in these subjects leads to resistance in many subjects. For example, one study of the use of sotorasib in the treatment of KRAS G12C-mutated colorectal cancer showed the about one-third of patients experienced early disease progression, exhibiting progression-free survival of less than 3 months. Thus, there is a need to develop new treatment paradigms for the treatment of KRAS G12C-mutated colorectal cancer.SUMMARY OF THE INVENTION

[0003] Disclosed herein are methods of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation.

[0004] Also disclosed herein are methods of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation by an FDA- approved test.

[0005] Further disclosed herein are methods of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0006] Also disclosed herein are methods of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation by an FDA approved test, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.WSGR Ref. 47134-784.601

[0007] Further disclosed herein are methods of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.INCORPORATION BY REFERENCE

[0008] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0009] FIG. 1 depicts the tumor growth inhibition (TGI) curves in animals comprising each treatment group (mean ± SEM) as set forth in Example 1, demonstrating the combination of adagrasib and Compound 1 resulted in the greatest TGI, the most tumor regressions and the most prolonged tumor growth control amongst the treatment groups.

[0010] FIG. 2 depicts tumor volumes from individual animals at day 18 in each of the treatment groups as set forth in Example 1, and the mean ± SEM for each group, demonstrating the combination of adagrasib plus Compound 1 resulted in a statistically significant reduction of tumor volumes. [Tukey’s multiple comparison, ****p<0.001]

[0011] FIG. 3 depicts tumor volumes from individual animals at day 42 from the adagrasib and adagrasib plus Compound 1 treatment groups as set forth in Example 1, and the mean ± SEM for each group, demonstrating the combination of adagrasib plus Compound 1 resulted in statistically significant smaller tumors. [***p<0.002, unpaired t-test] .DETAILED DESCRIPTION OF THE INVENTION

[0012] As used in the specification and appended claims, unless specified to the contrary, the following terms have the meaning indicated below.

[0013] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “an agent” includes a plurality of such agents, and reference to “the cell” includes reference to one or more cells (or to a plurality of cells) and equivalents thereof known to those skilled in the art, and so forth. When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formulae, all combinations and sub-combinations of ranges and specific embodiments therein are intended to be included. The term “about” when referring to a number or a numerical range means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range, in some instances, will vary between 1% and 15% of the stated number or numerical range. The term “comprising” (and related terms such as “comprise” or “comprises” or “having” or “including”) is not intended to exclude that in otherWSGR Ref. 47134-784.601certain embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, described herein, “consist of’ or “consist essentially of’ the described features.

[0014] “Administering” when used in conjunction with a therapeutic, including inhibitors of KRAS G12C and embryonic ectoderm development (EED) inhibitors, means to administer a therapeutic systemically or locally, as directly into or onto a target tissue, or to administer a therapeutic to a subject whereby the therapeutic positively impacts the tissue to which it is targeted. Thus, as used herein, the term “administering”, when used in conjunction with a composition described herein, can include, but is not limited to, providing a composition into or onto the target tissue; providing a composition systemically to a subject by, e.g., oral administration whereby the therapeutic reaches the target tissue or cells. “Administering” a composition may be accomplished by injection, topical administration, and oral administration or by other methods alone or in combination with other known techniques.

[0015] As used herein, the terms “subject,” “subject” and “individual” are intended to humans. In certain instances, the human is an adult. In certain instances, the human is child. In further instances, the human is under the age of 12 years. In certain instances, the human is elderly. In other instances, the human is 60 years of age or older. Other examples of subjects include experimental animals such as mice, rats, dogs, cats, goats, sheep, pigs, and cows. The experimental animal can be an animal model for a disorder, e.g., a transgenic mouse with hypertensive pathology.

[0016] The terms “determine,” “determined,” and “determining,” and the like, as used herein mean that it has been established that a pre-condition in a subject exists, or a condition precedent with respect to a subject has been satisfied, prior to the administration to the subject of inhibitors of KRAS G12C and embryonic ectoderm development (EED) inhibitors. For example, a biological sample from a subject having colorectal cancer (such as tumor tissue, blood, or plasma) is tested by methods known to those having ordinary skill in the art (such as by use of the QIAGEN therascreen KRAS RGQ PCR Kit) to determine whether the colorectal cancer in the subject comprises a KRAS G12C mutation prior to administration to the subject of (a) an inhibitor of KRAS G12C, and (b) an EED inhibitor. If the colorectal cancer in the subject is determined to comprise a KRAS G12C mutation, then the subject is administered the inhibitor of KRAS G12C and the EED inhibitor.

[0017] The term “embryonic ectoderm development (EED) inhibitor,” as used herein, means an agent that inhibits the function of the EED protein, including by binding to the protein.

[0018] The term “KRAS G12C,” as used herein, means a mutated form of the RAS GTPase protein KRAS in which the amino acid glycine (G) at position 12 of the amino acid sequence of the protein is replaced by cysteine (C).

[0019] The term “inhibitor of KRAS G12C,” as used herein means a molecule that exhibits inhibition of the activity of KRAS G12C and prevents its downstream signaling. It is specifically contemplated herein that inhibitors of KRAS G12C useful in the methods disclosed herein may exhibit inhibition of the KRAS G12C protein by any mechanism. KRAS G12C inhibitors that may be used in accordance with the methods disclosed herein include, but are not limited to, (a) molecules that bind directly to KRAS G12C, (b) pan-RAS inhibitors, and (c) RAS(ON) inhibitors. Such KRAS G12C inhibitors may be selected fromWSGR Ref. 47134-784.601the group comprising adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1 AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400 and RMC-4998. Inhibitors of KRAS G12C also include, but are not limited to, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, and YL-17231 (TEB-17231).

[0020] The term “pharmaceutically acceptable”, as used herein, means a carrier, diluent or excipient that is compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0021] The term “pharmaceutical composition” means a composition comprising at least one active ingredient, whereby the composition is amenable to investigation for a specified, efficacious outcome in a subject (for example, without limitation, a human). Those of ordinary skill in the art will understand and appreciate the techniques appropriate for determining whether an active ingredient has a desired efficacious outcome based upon the needs of the artisan.

[0022] As used herein, the term “therapeutic” means an agent utilized to treat, combat, ameliorate, prevent, or improve an unwanted condition or disease of a subject.

[0023] A “therapeutically effective amount” or “effective amount” as used herein refers to the amount of active compound or pharmaceutical agent that elicits a biological or medicinal response in a tissue, system, animal, individual or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of the following: (1) preventing the disease; for example, preventing a disease, condition or disorder in an individual that may be predisposed to the disease, condition or disorder but does not yet experience or display the pathology or symptomatology of the disease, (2) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and / or symptomatology), and (3) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual that is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and / or symptomatology).

[0024] The terms “treat,” “treated,” “treatment,” or “treating” as used herein refers to both therapeutic treatment in some embodiments and prophylactic or preventative measures in other embodiments, wherein the object is to prevent or slow (lessen) an undesired physiological condition, disorder, or disease, or to obtain beneficial or desired clinical results. For the purposes described herein, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), whetherWSGR Ref. 47134-784.601detectable or undetectable, or enhancement or improvement of the condition, disorder or disease.Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival as compared to expected survival if not receiving treatment. A prophylactic benefit of treatment includes prevention of a condition, retarding the progress of a condition, stabilization of a condition, or decreasing the likelihood of occurrence of a condition. For example, the efficacy of a given treatment regimen in a subject may be measured by criteria such as progression free survival (PFS), overall response rate (ORR), and / or the duration of response (DOR), all as measured according to criteria known to those having ordinary skill in the art such as RECIST vl.1.

[0025] For simplicity, chemical moi eties are defined and referred to throughout primarily as univalent chemical moi eties (e.g., alkyl, aryl, etc.). Nevertheless, such terms may also be used to convey corresponding multivalent moieties under the appropriate structural circumstances clear to those skilled in the art. For example, while an “alkyl” moiety generally refers to a monovalent radical (e.g. CH3-CH2-), in certain circumstances a bivalent linking moiety can be “alkyl,” in which case those skilled in the art will understand the alkyl to be a divalent radical (e.g. , -CH2-CH2-), which is equivalent to the term “alkylene.” (Similarly, in circumstances in which a divalent moiety is required and is stated as being “aryl,” those skilled in the art will understand that the term “aryl” refers to the corresponding divalent moiety, arylene.) All atoms are understood to have their normal number of valences for bond formation (i. e. , 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation state of the S).

[0026] The term “amino” as used herein refers to -NH2.

[0027] The term “acetyl” as used herein refers to “-C(O)CH3.

[0028] The term “acyl” as used herein refers to an alkylcarbonyl or arylcarbonyl substituent wherein the alkyl and aryl portions are as defined herein.

[0029] The term “alkyl” as used herein refers to straight and branched chain aliphatic groups having from 1 to 12 carbon atoms. As such, “alkyl” encompasses Ci, C2, C3, C4, Cs, Ce, C7, Cs, C9, C10, C11 and C12 groups. Examples of alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0030] The term “alkenyl” as used herein means an unsaturated straight or branched chain aliphatic group with one or more carbon-carbon double bonds, having from 2 to 12 carbon atoms. As such, “alkenyl” encompasses C2, C3, C4, Cs, Ce, C7, Cs, C9, C10, C11 and C12 groups. Examples of alkenyl groups include, without limitation, ethenyl, propenyl, butenyl, pentenyl, and hexenyl.

[0031] The term “alkynyl” as used herein means an unsaturated straight or branched chain aliphatic group with one or more carbon-carbon triple bonds, having from 2 to 12 carbon atoms. As such, “alkynyl” encompasses C2, C3, C4, Cs, Ce, C7, Cs, C9, C10, C11 and C12 groups. Examples of alkynyl groups include, without limitation, ethynyl, propynyl, butynyl, pentynyl, and hexynyl.

[0032] The terms “alkylene,” “alkenylene,” and “alkynylene” as used herein mean an alkyl, alkenyl, or alkynyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups. Examples of alkylene groups include, without limitation, methylene, ethylene, propylene, and butylene. Exemplary alkenylene groups include, without limitation, ethenylene,WSGR Ref. 47134-784.601propenylene, and butenylene. Exemplary alkynylene groups include, without limitation, ethynylene, propynylene, and butynylene.

[0033] The term “alkoxy” as used herein refers to -OCi-Ce alkyl.

[0034] The term “cycloalkyl” as used herein as employed herein is a saturated and partially unsaturated cyclic hydrocarbon group having 3 to 12 carbons. As such, “cycloalkyl” includes C3, C4, Cs, Ce, C7, Cs, C9, C10, Cn and C12 cyclic hydrocarbon groups. Examples of cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0035] The term “heteroalkyl” as used herein refers to an alkyl group, as defined hereinabove, wherein one or more carbon atoms in the chain are independently replaced by O, S, or NRX, wherein Rxis hydrogen or C1-C3 alkyl. Examples of heteroalkyl groups include methoxymethyl, methoxyethyl and methoxy propyl.

[0036] The term “aryl” as used herein means a Ce-C aromatic moiety comprising one to three aromatic rings. As such, “aryl” includes Ce, C10, C13, and C14 cyclic hydrocarbon groups. An exemplary aryl group is aCe-Cio aryl group. Particular aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, and fluorenyl.

[0037] The terms “aralkyl” and “arylalkyl” as used herein mean an aryl group covalently linked to an alkylene group wherein the moiety is linked to another group via the alkyl moiety. An exemplary aralkyl group is -(Ci-Ce)alkyl(C6-Cio)aryl, including, without limitation, benzyl, phenethyl, and naphthylmethyl.

[0038] The terms “heterocyclyl” and “heterocyclic” as used herein mean a mono- or bicyclic (fused or spiro) ring structure having from 3 to 12 atoms, (3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 atoms), for example 4 to 8 atoms, wherein one or more ring atoms are independently -C(O)-, N, NR5, O, or S, and the remainder of the ring atoms are quaternary or carbonyl carbons. Examples of heterocyclic groups include, without limitation, epoxy, oxiranyl, oxetanyl, azetidinyl, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, thiazolidinyl, thiatanyl, dithianyl, trithianyl, azathianyl, oxathianyl, dioxolanyl, oxazolidinyl, oxazolidinonyl, decahydroquinolinyl, piperidinyl, 4-piperidonyl, thiomorpholinyl, dimethyl -morpholinyl, and morpholinyl. Specifically excluded from the scope of this term are compounds having adjacent ring O and / or S atoms.

[0039] As used herein, term “L-heterocyclyl” as used herein means a heterocyclyl group covalently linked to another group via an alkylene linker L, where L is C1-C4 alkylene.

[0040] The term “heteroaryl” as used herein means a group having 5 to 14 ring atoms, preferably 5, 6, 10, 13 or 14 ring atoms comprising an aromatic heterocyclic ring (e.g., having 6, 10, or 1471 electrons shared in a cyclic array), and having, in addition to carbon atoms, from one to three heteroatoms that are each independently N, O, or S. “Heteroaryl” also includes fused multi cyclic (e.g. , bicyclic) ring systems in which one or more of the fused rings is non-aromatic, provided that at least one ring is aromatic and at least one ring contains an N, O, or S ring atom.WSGR Ref. 47134-784.601

[0041] Examples of heteroaryl groups include acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzo[d]oxazol-2(3H)-one, 2H-benzo[b][l,4]oxazin-3(4H)-one, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, furanyl, furazanyl, imidazolinyl, imidazolyl, IH-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-l,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl.

[0042] The terms “L-heteroaryl”, “heteroaralkyl” and “heteroarylalkyl” as used herein mean a group comprising a heteroaryl group covalently linked to another group via an alkylene linker. Examples of heteroalkyl groups comprise a Ci-Ce alkyl group and aheteroaryl group having 5, 6, 9, or 10 ring atoms. Examples of heteroaralkyl groups include pyridylmethyl, pyridylethyl, pyrrolylmethyl, pyrrolylethyl, imidazolylmethyl, imidazolylethyl, thiazolylmethyl, thiazolylethyl, benzimidazolylmethyl, benzimidazolylethyl quinazolinylmethyl, quinolinylmethyl, quinolinylethyl, benzofuranylmethyl, indolinylethyl isoquinolinylmethyl, isoinodylmethyl, cinnolinylmethyl, and benzothiophenylethyl. Specifically excluded from the scope of this term are compounds having adjacent ring O and / or S atoms.

[0043] The terms “arylene,” “heteroarylene,” and “heterocyclylene” as used herein mean an bivalent aryl, heteroaryl, or heterocyclyl group, respectively, as defined hereinabove, that is positioned between and serves to connect two other chemical groups.

[0044] As employed herein, when a moiety (e.g., cycloalkyl, aryl, heteroaryl, heterocyclyl, urea, etc.) is described as “optionally substituted” without expressly stating the substituents it is meant that the group optionally has from one to four, preferably from one to three, more preferably one or two, non -hydrogen substituents.

[0045] The terms “halogen” and “halo” as used herein mean chlorine, bromine, fluorine, or iodine.

[0046] The term “haloalkyl” as used herein means an alkyl chain in which one or more hydrogens have been replaced by a halogen. Exemplary haloalkyls are trifluoromethyl, difluoromethyl, fluorochloromethyl, chloromethyl, and fluoromethyl.

[0047] The term “hydroxyalkyl” as used herein means an alkyl chain, as defined herein, wherein at least one hydrogen of the alkyl chain has been replaced by hydroxyl.WSGR Ref. 47134-784.601

[0048] Disclosed herein are methods of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation.

[0049] Also disclosed herein are methods of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation by an FDA- approved test.

[0050] Further disclosed herein are methods of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0051] Also disclosed herein are methods of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation by an FDA approved test, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0052] Further disclosed herein are methods of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0053] In some embodiments, the subject disclosed herein has not acquired one or more secondary site pathway mutations. In some embodiments, the secondary site pathway mutation isa RAS / MAPK pathway alteration, optionally a KRAS alternation,a KRAS-activating mutation, optionally a mutation in codon 12 or 13,a KRAS amplification,a RAF / MAPK pathway mutation or fusion, optionally MAP2K1K57and BRAFV600E, a KRAS switch-II pocket mutation, optionally KRAS Y96D,an NRAS / HRAS mutation, optionally a single nucleotide variant in codon 61, a RTK amplification, capable of re-activating or mediating reactivation of a RAS / MAPK signaling pathway, optionally wherein the RTK amplification is MET amplification, ERBB2 amplification, ERBB4 amplification, EGFR amplification, or FGFR2 amplification,a RTK fusion, capable of re-activating or mediating reactivation of a RAS / MAPK signaling pathway, optionally wherein the RTK fusion is CCDC6-RET fusion, EML4-ALK fusion, FGFR3-TACC fusion, AHCYL2-MET fusion, MET-SLC12A9 fusion, or GOPC-ROS1 fusion, orany combination thereof.

[0054] In some embodiments, the KRAS-activating mutation is KRAS G12A, KRAS G12D, KRAS G12V, KRAS G13D, or KRAS Q61H; the KRAS switch-II pocket mutation is KRAS Y64 mutation, KRAS H95 mutation, KRAS Y96 mutation, optionally R68S, M72T, H95R, H95N, H95L, H95Q, Y96D, Y96N, Y96H, or Q99L; or the NRAS mutation is NRAS Q61K, or NRAS Q61R.WSGR Ref. 47134-784.601

[0055] In some embodiments are methods as disclosed herein wherein prior to the administration to the subject of the inhibitor of KRAS G12C and the EED inhibitor the subject has received one or more chemotherapeutic treatments. In some embodiments, the one or more chemotherapeutic treatments are selected from fluoropyrimidine-based chemotherapy, oxaliplatin-based chemotherapy, and irinotecanbased chemotherapy.

[0056] In some embodiments are provided methods of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, (b) an embryonic ectoderm development (EED) inhibitor, and (c) one or more anti-EGFR agents. In some embodiments are provided such methods wherein the one or more anti-EGFR agents are selected from EGFR antagonists, inhibitors, degraders, and antibody-drug conjugates. In still further embodiments are provided such methods wherein the one or more anti-EGFR agents are selected from panitumumab, cetuximab, osimertinib, erlotinib, gefitinib, lazertinib, firmonertinib, sunvozertinib, zipalertinib, ORIC-114, silevertinib (BDTX-1535), zanidatamab (ZW25), nimotuzumab, cetuximab saratolacan, depatuxizumab mafodotin, telisotuzumab vedotin (ABBV-399), AVID100, C225-ILs-dox, MRG003, E-EDV-D682, EGFR(V)-EDV-dox, EGR-ErbituxEDVsMIT, laptrituximab emtansine, losatuxizumab vedotin, serclutamab talirine, LD-DM1, R68-MC-VC-PAB-MMAE, RC68-PY-VC-PAB-MMAE, SHR-A1307, cetuximab-triptolide, M1231, and B2C4-MMAE.

[0057] In other embodiments are provided methods of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, RM-018, and RMC-4998. In some embodiments, the inhibitor of KRAS G12C is adagrasib or sotorasib. In some embodiments, the inhibitor of KRAS G12C is adagrasib. In some embodiments, the inhibitor of KRAS G12C is sotorasib. In some embodiments, the inhibitor of KRAS G12C is divarasib (GDC-6036). In some embodiments, the inhibitor of KRAS G12C is olomorasib (LY3537982). In some embodiments, the inhibitor of KRAS G12C is garsorasib (D-1553). In some embodiments, the inhibitor of KRAS G12C is glecirasib (JAB-21822). In some embodiments, the inhibitor of KRAS G12C is MK-1084. In some embodiments, the inhibitor of KRAS G12C is JNJ-74699157 (ARS-3248). In some embodiments, the inhibitor of KRAS G12C is GFH925. In some embodiments, the inhibitor of KRAS G12C is ZG19018. In some embodiments, the inhibitor of KRAS G12C is YL-15293. In some embodiments, the inhibitor of KRAS G12C is HBI-2438. In some embodiments, the inhibitor of KRAS G12C is FMC-376. In some embodiments, the inhibitor of KRAS G12C is HS-10370. In some embodiments, the inhibitor of KRAS G12C is BBO-8520. In some embodiments, the inhibitor of KRAS G12C is elironrasib (RMC-6291). In some embodiments, the inhibitor of KRAS G12C is KQB365. In some embodiments, the inhibitor ofWSGR Ref. 47134-784.601KRAS G12C is ARS-1620. In some embodiments, the inhibitor of KRAS G12C is 1_AM. In some embodiments, the inhibitor of KRAS G12C is ARS-853. In some embodiments, the inhibitor of KRAS G12C is BI-1823911. In some embodiments, the inhibitor of KRAS G12C is JDQ443. In some embodiments, the inhibitor of KRAS G12C is BPI-421286. In some embodiments, the inhibitor of KRAS G12C is GH35. In some embodiments, the inhibitor of KRAS G12C is BEBT-607. In some embodiments, the inhibitor of KRAS G12C is JAB-21000. In some embodiments, the inhibitor of KRAS G12C is LY3499446. In some embodiments, the inhibitor of KRAS G12C is SY-5933. In some embodiments, the inhibitor of KRAS G12C is HRS-7058. In some embodiments, the inhibitor of KRAS G12C is GEC255. In some embodiments, the inhibitor of KRAS G12C is daraxonrasib (RMC-6236). In some embodiments, the inhibitor of KRAS G12C is zoldonrasib (RMC-9805). In some embodiments, the inhibitor of KRAS G12C is RMC-7977. In some embodiments, the inhibitor of KRAS G12C is ADT-007. In some embodiments, the inhibitor of KRAS G12C is MRTX-1133. In some embodiments, the inhibitor of KRAS G12C is BI-2852. In some embodiments, the inhibitor of KRAS G12C is JAB-23400. In some embodiments, the inhibitor of KRAS G12C is RMC-4998.

[0058] In other embodiments, the inhibitor of KRAS G12C is selected from D35- 001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF -07985045, QTX3034, QTX3544, LUNA18, and YL-17231 (TEB-17231). In some embodiments, the inhibitor of KRAS G12C is D35-001. In some embodiments, the inhibitor of KRAS G12C is fulzerasib (GFH925, IBI351). In some embodiments, the inhibitor of KRAS G12C is opnurasib (JDQ-443). In some embodiments, the inhibitor of KRAS G12C is HYP-2090PTSA. In some embodiments, the inhibitor of KRAS G12C is BI-3706674. In some embodiments, the inhibitor of KRAS G12C is LY4066434. In some embodiments, the inhibitor of KRAS G12C is ALTA-3263. In some embodiments, the inhibitor of KRAS G12C is PF-07934040. In some embodiments, the inhibitor of KRAS G12C is BGB-53038. In some embodiments, the inhibitor of KRAS G12C is PF-07985045. In some embodiments, the inhibitor of KRAS G12C is QTX3034. In some embodiments, the inhibitor of KRAS G12C is QTX3544. In some embodiments, the inhibitor of KRAS G12C is LUNA18. In some embodiments, the inhibitor of KRAS G12C is YL-17231 (TEB-17231).

[0059] In another embodiment are any of the methods of treating colorectal cancer disclosed herein, wherein the EED inhibitor is a small molecule having a molecular weight of less or equal to 3000 Daltons. In some embodiments, the EED inhibitor is a small molecule having a molecular weight of less than or equal to 2000 Daltons, or less than or equal to 1500 Daltons, or less than or equal to 1000 Daltons, or less than or equal to 900 Daltons, or less than or equal to 800 Daltons, or less than or equal to 750 Daltons, or less than or equal to 700 Daltons, or less than or equal to 650 Daltons, or less than or equal to 600 Daltons, or less than or equal to 575 Daltons, or less than or equal to 525 Daltons, or less than or equal to 500 Daltons, or less than or equal to 475 Daltons, or less than or equal to 450 Daltons, or less than or equal to 425 Daltons, or less than or equal to 400 Daltons, or less than or equal to 375 Daltons, or less than or equal to 350 Daltons, or less than or equal to 325 Daltons, or less than or equal to 300WSGR Ref. 47134-784.601Daltons, or less than or equal to 275 Daltons, or less than or equal to 250 Daltons, or less than or equal to 200 Daltons.

[0060] In another embodiment are any of the methods of treating colorectal cancer disclosed herein, wherein the EED inhibitor is selected from EED226, A-395, APG-5918, BR-001, BR-002, EEDi-5285, EEDi-1056, pociredir (FTX-6058), HJM-353, and MAK683. In some embodiments, the EED inhibitor is EED226. In some embodiments, the EED inhibitor is A-395. In some embodiments, the EED inhibitor is APG-5918. In some embodiments, the EED inhibitor is BR-001. In some embodiments, the EED inhibitor is BR-002. In some embodiments, the EED inhibitor is EEDi-5285. In some embodiments, the EED inhibitor is EEDi-1056. In some embodiments, the EED inhibitoris pociredir (FTX-6058). In some embodiments, the EED inhibitor is HJM-353. In some embodiments, the EED inhibitor is MAK683.

[0061] In another embodiment are any of the methods of treating colorectal cancer disclosed herein, wherein the EED inhibitor is a compound of Formula (I)R1Formula (I)or a pharmaceutically acceptable salt thereof:wherein:represents a single or a double bond;Z is O or S;X is O, CR5, CR5OH, or C(R5)2, wherein:when X is O, is a single bond;when X is C(R5)2, is a single bond;when X is CR5OH, - is a single bond; orwhen X is CR5, is a double bond;R1is aryl, heteroaryl, L-cycloalkyl, -N(R5)heterocyclyl, or L-heterocyclyl, wherein the aryl, the heteroaryl or the cyclyl portion of the L-cycloalkyl, -N(R5)heterocyclyl, or L-heterocyclyl is optionally substituted with one or more R4;R2is cyano, -COOR5, -C(O)N(R5)2, or -C(O)N(R5)2 wherein each R5taken together with the nitrogen atom to which they are attached form a 5 - 8 membered heterocyclic ring optionally substituted with one or more R4;each R3is independently C1-C3 alkyl or halogen;each R4is independently oxo, cyano, halogen, -PCLCCi-Cs alkyl)2, hydroxyl, alkoxy, hydroxyalkyl, heteroalkyl, aralkyl, haloalkyl, -COOR5, -Y2-haloalkyl, -Y^Ci-Ce alkyl, -Y2-Ci-CeWSGR Ref. 47134-784.601alkyl, -L-cycloalkyl, -L-heteroaryl, -L-heterocyclyl, -Y'-heterocyclyl. -Y2-heterocyclyl, -L- N(R5)2, -O-L-N(R5)2, -C(CF3)N(R5)2, -Y1-N(R5)2, or -Y2-N(R5)2, wherein the ring portion of the aralkyl, -L-cycloalkyl, -L-heteroaryl, -L-heterocyclyl or -Y'-heterocycl l is optionally substituted with one or more R7;L is a bond or C1-C4 alkylene;Y1is a bond, -C(O)-, or -NHC(O)-;Y2is a bond, -S-, -SO-, -SO2-, or -NR5SO2-,each R5is hydrogen or C1-C3 alkyl;R6is hydrogen, C1-C3 alkyl, halogen, haloalkyl, hydroxyalkyl, or heteroalkyl;each R7is oxo, cyano, hydroxyl, alkoxy, halogen, haloalkyl, hydroxyalkyl, heteroalkyl, cycloalkyl, -L-N(R5)2, Ci-Ce alkyl, or -Y'-heterocyclyl; andn is 1 or 2.

[0062] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein Z is O. In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein Z is S.

[0063] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein n is 1.

[0064] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R2is cyano. In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R2is -COOR5. In some embodiments, the EED inhibitor is a compound of F ormula (I), or a pharmaceutically acceptable salt thereof, wherein R2is -C(O)N(R5)2.

[0065] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R3is halogen. In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R3is fluorine.

[0066] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein X is C(R5)2 and is a single bond.

[0067] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein X is CR5and is a double bond.

[0068] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein X is O and is a single bond.

[0069] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is aryl optionally substituted with one or more R4. In some embodiments, R1is phenyl optionally substituted with one or more R4. In some embodiments, R1is phenyl is substituted with one, two or three R4. In some embodiments, the one, two or three R4are each independently halogen, -PO3(Ci-C3 alkyl)2, hydroxyl, hydroxyallyl, aralkyl, haloalkyl, -COOR5, -Y^Ci-C6alkyl, Y2-Ci-C6alkyl, -L-N(R5)2, -O-L-N(R5)2, -C(CF3)N(R5)2, -Y!-N(R5)2, - 2-N(R5)2, Y2-haloalkyl, -L-heteroaryl, -L-heterocyclyl, or-Y1-heterocyclyl, whereintheheterocyclyl portion of the -L-heterocyclylWSGR Ref. 47134-784.601or -Y1-heterocyclyl is optionally substituted with one or more R7. In some embodiments, R4is -Y^Ci-Ce alkyl and Y1is a bond and the Ci-Ce alkyl is methyl, ethyl, isopropyl, butyl, or pentyl. In some embodiments, R4is -Y2-Ci-Ce alkyl and Y2is a -SO2- and the Ci-Ce alkyl is methyl. In some embodiments, R4is -Y2-haloalkyl and Y2is -S- or -SO2- and the haloalkyl is trifluoromethyl. In some embodiments, R4is -L-N(R5)2 and L is a bond and each R5is hydrogen, each R5is methyl, or one R5is methyl and one R5is hydrogen. In some embodiments, R4is -L-N(R5)2 and L is methylene or ethylene and each R5is hydrogen, each R5is methyl or one R5is methyl and one R5is hydrogen. In some embodiments, R4is -Y1-N(R5)2, Y1is -C(O)- and each R5independently is hydrogen, each R5is independently methyl or one R5is methyl and one R5is hydrogen. In some embodiments, R4is -Y2-N(R5)2, Y2is -SO2- and each R5independently is hydrogen, each R5is methyl, or one R5is methyl and one R5is independently hydrogen. In some embodiments, R4is -Y1-heterocyclyl and Y1is -C(O)- and the heterocyclyl portion of the L-heterocyclyl is piperazinyl or 4-methyl-piperazinyl. In some embodiments, R4is -L-heterocyclyl and L is a bond and the heterocyclyl portion of the L-heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or 3Z2-azabicyclo|3.1.0]hexanyl, each optionally substituted with one or more R7selected from oxo, C1-C3 alkyl, alkoxy, hydroxyl and halogen. In some embodiments, R4is -L-heterocyclyl, wherein Lis a methylene and the heterocyclyl portion of the L-heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl piperidinyl, each optionally substituted with one or more R7selected from C1-C3 alkyl, alkoxy, hydroxyl and halogen. In some embodiments, R4is -Y1-heterocyclyl and Y1is -C(O)- and the heterocyclyl portion of the Y1-heterocyclyl is morpholinyl optionally substituted with one or more C1-C3 alkyl. In some embodiments, R4is -L-heteroaryl optionally substituted with one or more R7. In some embodiments, R4is tetrazolyl. In some embodiments, R4is -PO3(Ci-C3 alkyl)2. In some embodiments, R4is -COOR5. In some embodiments, R4is hydroxyalkyl. In some embodiments, R4is -O-L-N(R5)2. In some embodiments, R4is aralkyl.

[0070] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is heteroaryl optionally substituted with one or more R4. In some embodiments, R1ispyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazinyl, pyridyl, pyridinyl-2-one, pyrazinyl, pyridazinyl, pyrimidinyl, isoxazolyl, isoindolinyl, naphthyridinyl, 1, 2,3,4-tetrahydroisoquinolinyl, or 5,6-dihydro-4H-pyrrolo[l,2-b]pyrazolyl, each optionally substituted with one or more R4. In some embodiments, R1is substituted with one or more R4; wherein each R4is independently cyano, halogen, -Y^Ci-Ce alkyl, -Y2-Ci-Ce alkyl, alkoxy, hydroxyalkyl, heteroalkyl, haloalkyl, -L-cycloalkyl, -L-N(R5)2,-Y1-N(R5)2, -L-heteroaryl, -L-heterocyclyl, or -Y1-heterocyclyl, wherein the heteroaryl of the -L-heteroaryl or the heterocyclyl portion of the L-heterocyclyl, or Y1-heterocyclyl is optionally substituted with one or more R7.

[0071] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is pyrazolyl optionally substituted with one R4independently selected from hydroxyalkyl, heteroalkyl, haloalkyl, -Y^Ci-Ce alkyl, -L-N(R5)2, L-heterocyclyl or L-heteroaryl, wherein the heteroaryl of the L-heteroaryl or the heterocyclyl portion of the L-heterocyclyl is optionally substituted with one or more R7. In some embodiments, R4is -L-heteroaryl and L is methylene whereinWSGR Ref. 47134-784.601the heteroaryl is pyridyl optional substituted with one or more R7. In some embodiments, R4is -L-heterocyclyl optionally substituted with one or more R7where Lisa bond and the heterocyclyl portion of the L-heterocyclyl is oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperazinyl, or 4-methylpiperazinyl. In some embodiments, R4is -L-heterocyclyl optionally substituted with one or more R7where L is methylene and the heterocyclyl portion of the L-heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl, pyrrolidinone, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, piperazinyl, or 4-methylpiperazinyl. In some embodiments, R4is -L-N(R5)2 where L is methylene and each R5is independently hydrogen, each R5is independently C1-C3 alkyl or one R5is C1-C3 alkyl and one R5is hydrogen. In some embodiments, R4is -Y1-CI-C6 alkyl where Y1is a bond and the Ci-Ce alkyl is methyl, ethyl, or isopropyl.

[0072] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is pyrazolyl optionally substituted with two R4groups each independently selected from hydroxyalkyl, heteroalkyl, haloalkyl, and -Y^Ci-Ce alkyl.

[0073] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is pyridyl optionally substituted with one R4independently selected from cyano, halogen, alkoxy, hydroxyalkyl, heteroalkyl, haloalkyl, -Y^Ci-Ce alkyl, -L-N(R5)2, -Y1-N(R5)2, -L-cycloalkyl, or -L-heterocyclyl optionally substituted with one or more R7.

[0074] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is -L-cycloalkyl optionally substituted with one or more R4.

[0075] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein R1is -L-heterocyclyl optionally substituted with one or more R4.

[0076] In some embodiments, the EED inhibitor is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, wherein L is a bond and the heterocyclyl is piperidinyl or tetrahydropyranyl.

[0077] In some embodiments, the EED inhibitor is a compound selected from the group consisting of:WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601

[0078] In some embodiments, the EED inhibitor is a compound selected from the group consisting of:acceptable salt thereof.WSGR Ref. 47134-784.601Q''NH

[0080] In some embodiments, the EED inhibitor is:O or a pharmaceutically acceptable salt thereof.

[0081] In some embodiments, the EED inhibitor is:or a pharmaceutically acceptable salt thereof.

[0082] In some embodiments, the EED inhibitor is:pharmaceutically acceptable salt thereof.

[0083] In some embodiments, the EED inhibitor is:° , or a pharmaceutically acceptable salt thereof.WSGR Ref. 47134-784.601

[0084] In some embodiments, the EED inhibitor is:acceptable salt thereof.

[0085] In some embodiments, the EED inhibitor is:acceptable salt thereof.

[0086] In some embodiments, the EED inhibitor is:acceptable salt thereof.

[0087] In some embodiments, the EED inhibitor is:acceptable salt thereof.WSGR Ref. 47134-784.601

[0088] In some embodiments, the EED inhibitor is:o , or a pharmaceutically acceptable salt thereof.

[0089] In some embodiments, the EED inhibitor is:0, or a pharmaceutically acceptable salt thereof.

[0090] In some embodiments, the EED inhibitor is:, or a pharmaceutically acceptable salt thereof.

[0091] Also provided herein are methods of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development(EED) inhibitor of formula° - (Compound 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasibWSGR Ref. 47134-784.601(RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, RMC-4998, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, and YL-17231 (TEB-17231). In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, and RMC-4998. In some embodiments, the inhibitor of KRAS G12C is adagrasib or sotorasib. In some embodiments, the inhibitor of KRAS G12C is adagrasib. In some embodiments, the inhibitor of KRAS G12C is sotorasib. In some embodiments, the inhibitor of KRAS G12C is divarasib (GDC-6036). In some embodiments, the inhibitor of KRAS G12C is olomorasib (LY3537982). In some embodiments, the inhibitor of KRAS G12C is garsorasib (D-1553). In some embodiments, the inhibitor of KRAS G12C is glecirasib (JAB-21822). In some embodiments, the inhibitor of KRAS G12C is MK-1084. In some embodiments, the inhibitor of KRAS G12C is JNJ-74699157 (ARS-3248). In some embodiments, the inhibitor of KRAS G12C is GFH925. In some embodiments, the inhibitor of KRAS G12C is ZG19018. In some embodiments, the inhibitor of KRAS G12C is YL-15293. In some embodiments, the inhibitor of KRAS G12C is HBI-2438. In some embodiments, the inhibitor of KRAS G12C is FMC-376. In some embodiments, the inhibitor of KRAS G12C is HS-10370. In some embodiments, the inhibitor of KRAS G12C is BBO-8520. In some embodiments, the inhibitor of KRAS G12C is elironrasib (RMC-6291). In some embodiments, the inhibitor of KRAS G12C is KQB365. In some embodiments, the inhibitor of KRAS G12C is ARS-1620. In some embodiments, the inhibitor of KRAS G12C is 1_AM. In some embodiments, the inhibitor of KRAS G12C is ARS-853. In some embodiments, the inhibitor of KRAS G12C is BI-1823911. In some embodiments, the inhibitor of KRAS G12C is JDQ443. In some embodiments, the inhibitor of KRAS G12C is BPI-421286. In some embodiments, the inhibitor of KRAS G12C is GH35. In some embodiments, the inhibitor of KRAS G12C is BEBT-607. In some embodiments, the inhibitor of KRAS G12C is JAB-21000. In some embodiments, the inhibitor of KRAS G12C is LY3499446. In some embodiments, the inhibitor of KRAS G12C is SY-5933. In some embodiments, the inhibitor of KRAS G12C is HRS-7058. In some embodiments, the inhibitor of KRAS G12C is GEC255. In some embodiments, the inhibitor of KRAS G12C is daraxonrasib (RMC-6236). In some embodiments, the inhibitor of KRAS G12C is zoldonrasib (RMC-9805). In some embodiments, the inhibitor of KRAS G12C is RMC-7977. In some embodiments, the inhibitor of KRAS G12C is ADT-007. In some embodiments, the inhibitor of KRAS G12C is MRTX-1133. In some embodiments, the inhibitor of KRAS G12C is BI-2852. In some embodiments, the inhibitor of KRAS G12C is JAB-23400. In some embodiments, the inhibitor of KRAS G12C is RMC-4998.WSGR Ref. 47134-784.601

[0092] Further provided herein are methods of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development INH(EED) inhibitor of formula <x J (Compound 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, RMC-4998, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, and YL-17231 (TEB-17231). In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, and RMC-4998. In some embodiments, the inhibitor of KRAS G12C is adagrasib or sotorasib. In some embodiments, the inhibitor of KRAS G12C is adagrasib. In some embodiments, the inhibitor of KRAS G12C is sotorasib. In some embodiments, the inhibitor of KRAS G12C is divarasib (GDC-6036). In some embodiments, the inhibitor of KRAS G12C is olomorasib (LY3537982). In some embodiments, the inhibitor of KRAS G12C is garsorasib (D-1553). In some embodiments, the inhibitor of KRAS G12C is glecirasib (JAB-21822). In some embodiments, the inhibitor of KRAS G12C is MK-1084. In some embodiments, the inhibitor of KRAS G12C is JNJ-74699157 (ARS-3248). In some embodiments, the inhibitor of KRAS G12C is GFH925. In some embodiments, the inhibitor of KRAS G12C is ZG19018. In some embodiments, the inhibitor of KRAS G12C is YL-15293. In some embodiments, the inhibitor of KRAS G12C is HBI-2438. In some embodiments, the inhibitor of KRAS G12C is FMC-376. In some embodiments, the inhibitor of KRAS G12C is HS-10370. In some embodiments, the inhibitor of KRAS G12C is BBO-8520. In some embodiments, the inhibitor of KRAS G12C is elironrasib (RMC-6291). In some embodiments, the inhibitor of KRAS G12C is KQB365. In some embodiments, the inhibitor of KRAS G12C is ARS-1620.WSGR Ref. 47134-784.601In some embodiments, the inhibitor of KRAS G12C is 1_AM. In some embodiments, the inhibitor of KRAS G12C is ARS-853. In some embodiments, the inhibitor of KRAS G12C is BI-1823911. In some embodiments, the inhibitor of KRAS G12C is JDQ443. In some embodiments, the inhibitor of KRAS G12C is BPI-421286. In some embodiments, the inhibitor of KRAS G12C is GH35. In some embodiments, the inhibitor of KRAS G12C is BEBT-607. In some embodiments, the inhibitor of KRAS G12C is JAB-21000. In some embodiments, the inhibitor of KRAS G12C is LY3499446. In some embodiments, the inhibitor of KRAS G12C is SY-5933. In some embodiments, the inhibitor of KRAS G12C is HRS-7058. In some embodiments, the inhibitor of KRAS G12C is GEC255. In some embodiments, the inhibitor of KRAS G12C is daraxonrasib (RMC-6236). In some embodiments, the inhibitor of KRAS G12C is zoldonrasib (RMC-9805). In some embodiments, the inhibitor of KRAS G12C is RMC-7977. In some embodiments, the inhibitor of KRAS G12C is ADT-007. In some embodiments, the inhibitor of KRAS G12C is MRTX-1133. In some embodiments, the inhibitor of KRAS G12C is BI-2852. In some embodiments, the inhibitor of KRAS G12C is JAB-23400. In some embodiments, the inhibitor of KRAS G12C is RMC-4998.

[0093] Also provided herein are methods of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation by an FDA approved test, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm INHdevelopment (EED) inhibitor of formulanj (Compound 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, RMC-4998, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, and YL-17231 (TEB-17231). In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007,WSGR Ref. 47134-784.601MRTX-1133, BI-2852, JAB-23400, andRMC-4998. In some embodiments, the inhibitor of KRAS G12C is adagrasib or sotorasib. In some embodiments, the inhibitor of KRAS G12C is adagrasib. In some embodiments, the inhibitor of KRAS G12C is sotorasib. In some embodiments, the inhibitor of KRAS G12C is divarasib (GDC-6036). In some embodiments, the inhibitor of KRAS G12C is olomorasib (LY3537982). In some embodiments, the inhibitor of KRAS G12C is garsorasib (D-1553). In some embodiments, the inhibitor of KRAS G12C is glecirasib (JAB-21822). In some embodiments, the inhibitor of KRAS G12C is MK-1084. In some embodiments, the inhibitor of KRAS G12C is JNJ-74699157 (ARS-3248). In some embodiments, the inhibitor of KRAS G12C is GFH925. In some embodiments, the inhibitor of KRAS G12C is ZG19018. In some embodiments, the inhibitor of KRAS G12C is YL-15293. In some embodiments, the inhibitor of KRAS G12C is HBI-2438. In some embodiments, the inhibitor of KRAS G12C is FMC-376. In some embodiments, the inhibitor of KRAS G12C is HS-10370. In some embodiments, the inhibitor of KRAS G12C is BBO-8520. In some embodiments, the inhibitor of KRAS G12C is elironrasib (RMC-6291). In some embodiments, the inhibitor of KRAS G12C is KQB365. In some embodiments, the inhibitor of KRAS G12C is ARS-1620. In some embodiments, the inhibitor of KRAS G12C is 1_AM. In some embodiments, the inhibitor of KRAS G12C is ARS-853. In some embodiments, the inhibitor of KRAS G12C is BI-1823911. In some embodiments, the inhibitor of KRAS G12C is JDQ443. In some embodiments, the inhibitor of KRAS G12C is BPI-421286. In some embodiments, the inhibitor of KRAS G12C is GH35. In some embodiments, the inhibitor of KRAS G12C is BEBT-607. In some embodiments, the inhibitor of KRAS G12C is JAB-21000. In some embodiments, the inhibitor of KRAS G12C is LY3499446. In some embodiments, the inhibitor of KRAS G12C is SY-5933. In some embodiments, the inhibitor of KRAS G12C is HRS-7058. In some embodiments, the inhibitor of KRAS G12C is GEC255. In some embodiments, the inhibitor of KRAS G12C is daraxonrasib (RMC-6236). In some embodiments, the inhibitor of KRAS G12C is zoldonrasib (RMC-9805). In some embodiments, the inhibitor of KRAS G12C is RMC-7977. In some embodiments, the inhibitor of KRAS G12C is ADT-007. In some embodiments, the inhibitor of KRAS G12C is MRTX-1133. In some embodiments, the inhibitor of KRAS G12C is BI-2852. In some embodiments, the inhibitor of KRAS G12C is JAB-23400. In some embodiments, the inhibitor of KRAS G12C is RMC-4998.

[0094] Further provided herein are methods of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm INHz^^sk / F< X Jdevelopment (EED) inhibitor of formula(Compound 1), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of KRAS G12C is selected from adagrasib,WSGR Ref. 47134-784.601sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, RMC-4998, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, and YL-17231 (TEB-17231). In some embodimenls, the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, and RMC-4998In some embodiments, the inhibitor of KRAS G12C is adagrasib or sotorasib. In some embodiments, the inhibitor of KRAS G12C is adagrasib. In some embodiments, the inhibitor of KRAS G12C is sotorasib. In some embodiments, the inhibitor of KRAS G12C is divarasib (GDC-6036). In some embodiments, the inhibitor of KRAS G12C is olomorasib (LY3537982). In some embodiments, the inhibitor of KRAS G12C is garsorasib (D-1553). In some embodiments, the inhibitor of KRAS G12C is glecirasib (JAB-21822). In some embodiments, the inhibitor of KRAS G12C is MK-1084. In some embodiments, the inhibitor of KRAS G12C is JNJ-74699157 (ARS-3248). In some embodiments, the inhibitor of KRAS G12C is GFH925. In some embodiments, the inhibitor of KRAS G12C is ZG19018. In some embodiments, the inhibitor of KRAS G12C is YL-15293. In some embodiments, the inhibitor of KRAS G12C is HBI-2438. In some embodiments, the inhibitor of KRAS G12C is FMC-376. In some embodiments, the inhibitor of KRAS G12C is HS-10370. In some embodiments, the inhibitor of KRAS G12C is BBO-8520. In some embodiments, the inhibitor of KRAS G12C is elironrasib (RMC-6291). In some embodiments, the inhibitor of KRAS G12C is KQB365. In some embodiments, the inhibitor of KRAS G12C is ARS-1620. In some embodiments, the inhibitor of KRAS G12C is 1_AM. In some embodiments, the inhibitor of KRAS G12C is ARS-853. In some embodiments, the inhibitor of KRAS G12C is BI-1823911. In some embodiments, the inhibitor of KRAS G12C is JDQ443. In some embodiments, the inhibitor of KRAS G12C is BPI-421286. In some embodiments, the inhibitor of KRAS G12C is GH35. In some embodiments, the inhibitor of KRAS G12C is BEBT-607. In some embodiments, the inhibitor of KRAS G12C is JAB-21000. In some embodiments, the inhibitor of KRAS G12C is LY3499446. In some embodiments, the inhibitor of KRAS G12C is SY-5933. In some embodiments, the inhibitor of KRAS G12C is HRS-7058. In some embodiments, the inhibitor of KRAS G12C is GEC255. In some embodiments, the inhibitor of KRAS G12C is daraxonrasib (RMC-6236). In some embodiments, the inhibitor of KRAS G12C is zoldonrasib (RMC-9805). In some embodiments, the inhibitor of KRAS G12C is RMC-7977. In some embodiments, the inhibitor of KRAS G12C is ADT-007. In someWSGR Ref. 47134-784.601embodiments, the inhibitor of KRAS G12C is MRTX-1133. In some embodiments, the inhibitor of KRAS G12C is BI-2852. In some embodiments, the inhibitor of KRAS G12C is JAB-23400. In some embodiments, the inhibitor of KRAS G12C is RMC-4998.

[0095] Also provided herein are any of the methods of treating colorectal cancer in a subject disclosed herein, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) Compound 1:I>LIACNNH(Compound 1), or a pharmaceutically acceptable salt thereof, wherein Compound 1 is in crystalline form. Also provided herein are such methods wherein the crystalline form of Compound 1 is an anhydrous form. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 8.1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 9.6° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.7°± 0.2°2-theta, 19.7° ± 0.2° 2-theta, and 22.0° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 9.8° ± 0.2°2-theta, 15.2° ± 0.2° 2-theta, and 17.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of about 172 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 205 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C, or from about 207 °C to about 210 °C, or from about 208 °C to about 210 °C, or from about 209 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 1% upon heating the sample from about 25 °C to a temperature prior to melting. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 1% upon heating the sample from about 25 °C to about 380 °C.

[0096] Also provided herein are any of the methods disclosed herein wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta and 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ±WSGR Ref. 47134-784.6010.2° 2-theta, and 24.1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 15.4° ± 0.2° 2-theta, 21.0° ± 0.2° 2-theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0097] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further a peak in an x-ray powder diffraction (XRPD) pattern at 15.4° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further apeak in an x-ray powder diffraction (XRPD) pattern at 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2-theta, and 24.1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) pattern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 21.0° ± 0.2° 2-theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0098] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits peaks in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta and 15.4° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-rayWSGR Ref. 47134-784.601powder diffraction (XRPD) patern at 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) patern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2-theta, and 24.1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) patern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 21.0° ± 0.2° 2-theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry patern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0099] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits peaks in an x-ray powder diffraction (XRPD) patern at 7.7° ± 0.2° 2-theta and 19.2° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in an x-ray powder diffraction (XRPD) patern at 15.4° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a further peak in an x-ray powder diffraction (XRPD) pattern at 13.7° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) patern at 5.5° ± 0.2° 2-theta, 8.6° ± 0.2° 2-theta, 15.9° ± 0.2° 2-theta, 19.9° ± 0.2° 2-theta, and 24.1° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits further peaks in an x-ray powder diffraction (XRPD) patern at 10.6° ± 0.2° 2-theta, 11.0° ± 0.2° 2-theta, 21.0° ± 0.2° 2-theta, and 26.3° ± 0.2° 2-theta. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry patern of from about 203 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 206 °C to about 210 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a peak in a differential scanning calorimetry pattern of from about 203 °C to about 208 °C, or from about 203 °C to about 206 °C, or from about 203 °C to about 205 °C. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 380 °C. Also provided herein are such methods wherein theWSGR Ref. 47134-784.601crystalline form of Compound 1 exhibits a loss in mass in a thermal gravimetric analysis of less than about 2% upon heating the sample from about 25 °C to about 210 °C.

[0100] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 10% degradation when stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days.

[0101] Also provided herein are such methods wherein the crystalline form of Compound 1 (a) exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 8.1° ± 0.2° 2-theta, and (b) exhibits less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 (a) exhibits peaks in an x-ray powder diffraction (XRPD) pattern at 9.6°± 0.2° 2-theta, 5.7° ± 0.2° 2-theta, 19.7°± 0.2° 2-theta, and 22.0° ± 0.2° 2-theta , and (b) less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days.

[0102] Also provided herein are such methods wherein the crystalline form of Compound 1 (a) exhibits a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, and (b) exhibits less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) peaks in an x-ray powder diffraction (XRPD) pattern at 7.7°± 0.2° 2-theta, 13.7° ± 0.2° 2-theta, and 19.2° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 25 °C and 60% relative humidity for at least 7 days.

[0103] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 10% degradation when the crystalline forms are stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days.

[0104] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) a peak in an x-ray powder diffraction (XRPD) pattern at 8.1° ± 0.2° 2-theta, and (b) less than about 10%WSGR Ref. 47134-784.601degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) peaks in an x-ray powder diffraction (XRPD) pattern at 9.6°± 0.2° 2-theta, 5.7° ± 0.2° 2-theta, 19.7° ± 0.2° 2-theta, and 22.0°± 0.2° 2-theta , and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days.

[0105] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) a peak in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits (a) peaks in an x-ray powder diffraction (XRPD) pattern at 7.7° ± 0.2° 2-theta, 13.7° ± 0.2° 2-theta, and 19.2° ± 0.2° 2-theta, and (b) less than about 10% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 40 °C and 75% relative humidity for at least 7 days.

[0106] Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 10% degradation when the crystalline form is stored at 60 °C for at least one week. Also provided herein are such methods wherein the crystalline form of Compound 1 exhibits less than about 1%, or less than about 2%, or less than about 3%, or less than about 4%, or less than about 5%, or less than about 6%, or less than about 7%, or less than about 8%, or less than about 9% degradation when the crystalline form is stored at 60 °C for at least one week.

[0107] Also disclosed herein are any of the methods of treating colorectal cancer disclosed herein, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject sequentially or simultaneously. In an embodiment, the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject sequentially. In another embodiment, the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject simultaneously.

[0108] Also disclosed herein are any of the methods of treating colorectal cancer disclosed herein, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject on the same day.

[0109] Also disclosed herein are any of the methods of treating colorectal cancer disclosed herein, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject within a 24-hour period. In another embodiment, the inhibitor of KRAS G12C and the EED inhibitor are administeredWSGR Ref. 47134-784.601to the subject within a 12-hour period, or a 10-hour period, or an 8-hour period, or a 6-hour period, or a -hour period, or a 2-hour period, or within an hour of each of other.

[0110] Also disclosed herein are any of the methods of treating colorectal cancer disclosed herein, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject once per day or twice per day. In an embodiment, the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject once per day. In an embodiment, the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject twice per day. In an embodiment, the inhibitor of KRAS G12C is administered to the subject twice per day and the EED inhibitor is administered to the subject once per day. In an embodiment, the inhibitor of KRAS G12C is administered to the subject once per day and the EED inhibitor is administered to the subject twice per day.[OOlllJAlso disclosed herein are any of the methods of treating colorectal cancer disclosed herein, wherein the inhibitor of KRAS G12C is administered to the subject with food or without food. In an embodiment, the inhibitor of KRAS G12C is administered to the subject with food. In an embodiment, the inhibitor of KRAS G12C is administered to the subject without food.

[0112] In some embodiments of the methods of treating colorectal cancer disclosed herein the EED inhibitor is administered to the subject with or without food. In some embodiments, the EED inhibitor is administered to the subject with food. In some embodiments, the EED inhibitor is administered to the subject without food.

[0113] In some embodiments are provided any of the methods of treating colorectal cancer in a subject disclosed herein, wherein the colorectal cancer in the subject is selected from metastatic colorectal cancer, locally advanced colorectal cancer, locally advanced metastatic colorectal cancer, non-metastatic colorectal cancer, colon cancer, metastatic colon cancer, locally advanced colon cancer, locally advanced metastatic colon cancer, non-metastatic colon cancer, rectal cancer, metastatic rectal cancer, locally advanced rectal cancer, locally advanced metastatic rectal cancer, non-metastatic rectal cancer. In some embodiments, the colorectal cancer in the subject is metastatic colorectal cancer. In some embodiments, the colorectal cancer in the subject is locally advanced colorectal cancer. In some embodiments, the colorectal cancer in the subject is locally advanced metastatic colorectal cancer. In some embodiments, the colorectal cancer in the subject is non-metastatic colorectal cancer,. In some embodiments, the colorectal cancer in the subject is colon cancer. In some embodiments, the colorectal cancer in the subject is metastatic colon cancer. In some embodiments, the colorectal cancer in the subject is locally advanced colon cancer. In some embodiments, the colorectal cancer in the subject is locally advanced metastatic colon cancer. In some embodiments, the colorectal cancer in the subject is non- metastatic colon cancer. In some embodiments, the colorectal cancer in the subject is rectal cancer. In some embodiments, the colorectal cancer in the subject is metastatic rectal cancer. In some embodiments, the colorectal cancer in the subject is locally advanced rectal cancer. In some embodiments, the colorectal cancer in the subject is locally advanced metastatic rectal cancer. In some embodiments, the colorectal cancer in the subject is non-metastatic rectal cancer.WSGR Ref. 47134-784.601

[0114] The compounds of Formula (I), or pharmaceutically acceptable salts thereof, may be prepared using commercially available reagents and intermediates in the synthetic methods and reaction schemes described herein, those described in United States Patent No. 11,091,495, or may be prepared using other reagents and conventional methods well known to those skilled in the art. The contents of United States Patent No. 11,091,495 are hereby incorporated by reference for that purpose.

[0115] For instance, intermediates for compounds and compounds of Formula (I) of the present invention may be prepared according to General Reaction Schemes I or II:General Reaction Scheme I

[0116] In General Reaction Scheme I, R2-ester substituted imidazo[l,2-c]pyrimidine A is coupled to R3 optionally substituted intermediate amine B by nucleophilic substitution to yield Intermediate C. A boronic acid derivative (Y)-Rl D is coupled via a Suzuki reaction with halogen substituted Intermediate C in the presence of a suitable base, e.g., sodium carbonate, and the R2 ester is converted to the acid by saponification with NaOH to generate intermediate acid E. The acid is converted to the corresponding amide, which is dehydrated to form title compound nitrile G.General Reaction Scheme IIWSGR Ref. 47134-784.601Br R1C

[0117] In General Reaction Scheme II, halogenated Intermediate C containing a suitable R2 reactant, e.g., an ester, in the presence of a suitable base is converted to acid intermediate by saponification, then treated with NH4C1 in the presence of HATU to form the amide which is subsequently dehydrated to form nitrile Intermediate H. R1 is coupled to Intermediate H via a Suzuki reaction using boronic acid derivative (Y) in the presence of base. The nitrile group of R1 -containing Intermediate G is hydrolyzed in the presence of acid and water to afford title compound amide F.

[0118] In some embodiments are provided methods of treatment of colorectal cancer in a subject, comprising administering to the subject a pharmaceutically acceptable salt of the compounds of Formula (I). The desired salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid, such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like. It is specifically contemplated herein that references to the compounds of Formula (I), also refer in the alternative to pharmaceutically acceptable salts of compounds of Formula (I).

[0119] If the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is a solid, it is understood by those skilled in the art that the compounds or salts thereof may exist in different crystal or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas.

[0120] Also provided herein are uses of isotopically-labeled compounds of Formula (I), or a pharmaceutically acceptable thereof, wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of hydrogen, such as2H and3H, carbon, such as11C,13C and14C, chlorine, such as36C1,WSGR Ref. 47134-784.601fluorine, such as18F, iodine, such as123I and125I, nitrogen, such as13N and15N, oxygen, such as15O,17O and18O, phosphorus, such as32P, and sulfur, such as35S. Certain isotopically-labeled compounds of the invention, for example, those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The radioactive isotopes tritium (3H) and carbon-14 (14C) are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Substitution with heavier isotopes such as deuterium,2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Substitution with positron emitting isotopes, such asnC,18F,15O and13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds of Formula (I), or a pharmaceutically acceptable salt thereof, can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described herein, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent otherwise employed.

[0121] In one aspect, the inhibitor of KRAS G12C and the EED inhibitor may be administered to a subject having colorectal cancer as described herein in the form of pharmaceutically acceptable compositions. Such compositions may be prepared in pharmaceutically acceptable dosage forms for administration to subjects. Pharmaceutically acceptable dosage forms include, for example, liquids, suspensions, powders for reconstitution, tablets, pills, sachets, or capsules of hard or soft gelatin (See, e.g., Remington: The Science and Practice of Pharmacy (Gennaro, 21stEd. Mack Pub. Co., Easton, PA (2005)). In some embodiments, the inhibitor of KRAS G12C and the EED inhibitor may be formulated into pharmaceutical compositions as described below in any pharmaceutical form recognizable to the skilled artisan as being suitable. Such pharmaceutical compositions comprise a therapeutically effective amount of the inhibitor of KRAS G12C and / or the EED inhibitor, and an inert, pharmaceutically acceptable carrier or diluent. Such compositions may comprise both the inhibitor of KRAS G12C and the EED inhibitor, or the inhibitor of KRAS G12C and the EED inhibitor may be formulated separately for separate administration to the subject having colorectal cancer.

[0122] The pharmaceutical carriers employed may be either solid or liquid. Exemplary solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and the like.Exemplary liquid carriers are syrup, peanut oil, olive oil, water, and the like. Similarly, the compositions may include time-delay or time-release material known in the art, such as glyceryl monostearate or glyceryl distearate alone or with a wax, ethyl cellulose, hydroxypropylmethylcellulose, methylmethacrylate or the like. Further additives or excipients may be added to achieve the desired formulation properties. For example, a bioavailability enhancer, such as Labrasol, Gelucire or the like, or formulator, such as CMC (carboxy-methylcellulose), PG (propyleneglycol), or PEG (polyethyleneglycol), may be added. Gelucire, a semi-solid vehicle that protects active ingredients from light, moisture, and oxidation, may be added, e.g., when preparing a capsule formulation.

[0123] If a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form, or formed into a troche or lozenge. The amount of solid carrier may vary, butWSGR Ref. 47134-784.601generally will be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampoule or vial or non-aqueous liquid suspension. If a semi-solid carrier is used, the preparation may be in the form of hard and soft gelatin capsule formulations. The inventive compositions are prepared in unit-dosage form appropriate for the mode of administration, e.g. parenteral or oral administration.

[0124] To obtain a stable water-soluble dose form, a suitable compound may be dissolved in an aqueous solution of an organic or inorganic acid, such as a 0.3 M solution of succinic acid or citric acid. If a soluble salt form is not available, the compound, or a pharmaceutically acceptable salt thereof, may be dissolved in a suitable co-solvent or combinations of co-solvents. Examples of suitable co-solvents include alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerin and the like in concentrations ranging from 0 to 60% of the total volume. In an exemplary embodiment, a suitable compound is dissolved in DMSO and diluted with water. The composition may also be in the form of a solution of a salt form of the active ingredient in an appropriate aqueous vehicle such as water or isotonic saline or dextrose solution.

[0125] Proper formulation is dependent upon the route of administration selected. For injection, a suitable compound may be formulated into aqueous solutions, preferably in physiologically compatible buffers such as Hanks solution, Ringer's solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0126] For oral administration, a suitable compound can be formulated by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a subject to be treated. Pharmaceutical preparations for oral use can be obtained using a solid excipient in admixture with the active ingredient (agent), optionally grinding the resulting mixture, and processing the mixture of granules after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include: fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as crosslinked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0127] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinyl pyrrolidone, Carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active agents.

[0128] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fitWSGR Ref. 47134-784.601capsules can contain the active ingredients in admixture with fillers such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0129] For administration intranasally or by inhalation, a suitable compound may be conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, tri chlorofluoromethane, di chlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of gelatin for use in an inhaler or insufflator and the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0130] The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit-dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0131] Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active agents may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[0132] Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile pyrogen-free water, before use.

[0133] In addition to the formulations described above, a suitable compound may also be formulated as a depot preparation. Such long-acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion-exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt. A pharmaceutical carrier for hydrophobic compounds is a co-solvent system comprising benzyl alcohol, anon-polar surfactant, a water-miscible organic polymer, and an aqueous phase. The cosolvent system may be a VPD co-solvent system. VPD is a solution of 3% w / v benzyl alcohol, 8% w / v of the non-polar surfactant polysorbate 80, and 65% w / v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD: 5W) contains VPD diluted 1:1 with a 5% dextroseWSGR Ref. 47134-784.601in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. The proportions of a co-solvent system may be suitably varied without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity non-polar surfactants may be used instead of polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars or polysaccharides may be substituted for dextrose.

[0134] Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide (DMSO) also may be employed, although usually at the cost of greater toxicity due to the toxic nature of DMSO. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.

[0135] The pharmaceutical compositions also may comprise suitable solid- or gel-phase carriers or excipients. These carriers and excipients may provide marked improvement in the bioavailability of poorly soluble drugs. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0136] Further, the pharmaceutical composition may be incorporated into a skin patch for delivery of the drug directly onto the skin.

[0137] Additionally, the pharmaceutically acceptable compositions that may be used to practice the methods disclosed herein may contain a suitable compound in an amount of from about 0.5 w / w % to about 95 w / w %, or from about 1 w / w % to about 95 w / w %, or from about 1 w / w % to about 75 w / w %, or from about 5 w / w % to about 75 w / w %, or from about 10 w / w % to about 75 w / w %, or from about 10 w / w % to about 50 w / w %.

[0138] It will be appreciated that the actual dosages of the compound of Formula (I), or a pharmaceutically acceptable salt thereof, to be administered to a subject in need thereof, will vary according to the particular agent being used, the particular composition formulated, the mode of administration, and the particular site, host, and disease being treated. Those skilled in the art using conventional dosage-determination tests in view of the experimental data for a given compound may ascertain optimal dosages for a given set of conditions. For oral administration, an exemplary daily dose generally employed will be from about 0.001 to about 1000 mg / kg of body weight, with courses of treatment repeated at appropriate intervals. In some embodiments are provided the methods disclosed herein, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered to the subject in an amount between about 0.01 mg / kg per day to about 300 mg / kg per day.WSGR Ref. 47134-784.601In other embodiments are provided the methods disclosed herein, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered to the subject in an amount between about 0.1 mg / kg per day to about 100 mg / kg per day.

[0139] In some embodiments are provided the methods disclosed herein, wherein the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered to the subject in an amount between about 100 mg and about 1000 mg once per day, between about 100 mg and about 900 mg once per day, between about 100 mg and about 850 mg once per day, between about 100 mg and about 800 mg once per day, between about 100 mg and about 750 mg once per day, between about 100 mg and about 700 mg once per day, between about 100 mg and about 650 mg once per day, between about 100 mg and about 600 mg once per day, between about 100 mg and about 550 mg once per day, or between about 100 mg and about 500 mg once per day.

[0140] In some embodiments are provided the methods disclosed herein, wherein the compound of F ormula (I), or a pharmaceutically acceptable salt thereof, is administered to the subj ect in an amount of about 100 mg once per day, about 150 mg once per day, about 200 mg once per day, about 300 mg once per day, about 225 mg once per day, about 275 mg once per day, about 300 mg once per day, about 325 mg once per day, about 350 mg once per day, about 375 mg once per day, about 400 mg once per day, about 425 mg once per day, about 450 mg once per day, about 475 mg once per day, about 500 mg once per day, about 525 mg once per day, about 550 mg once per day, about 575 mg once per day, about 600 mg once per day, about 625 mg once per day, about 650 mg once per day, about 675 mg once per day, about 700 mg once per day, about 725 mg once per day, about 750 mg once per day, about 775 mg once per day, about 800 mg once per day, about 825 mg once per day, about 850 mg once per day, about 875 mg once per day, about 900 mg once per day, about 925 mg once per day, about 950 mg once per day, about 975 mg once per day, or about 1000 mg once per day.

[0141] In some embodiments are provided the methods disclosed herein, wherein the Compound 1 is administered to the subject in an amount between about 100 mg and about 1000 mg once per day, between about 100 mg and about 900 mg once per day, between about 100 mg and about 850 mg once per day, between about 100 mg and about 800 mg once per day, between about 100 mg and about 750 mg once per day, between about 100 mg and about 700 mg once per day, between about 100 mg and about 650 mg once per day, between about 100 mg and about 600 mg once per day, between about 100 mg and about 550 mg once per day, or between about 100 mg and about 500 mg once per day.

[0142] In some embodiments are provided the methods disclosed herein, wherein the Compound 1 is administered to the subject in an amount of about 100 mg once per day, about 150 mg once per day, about 200 mg once per day, about 225 mg once per day, about 275 mg once per day, about 300 mg once per day, about 325 mg once per day, about 350 mg once per day, about 375 mg once per day, about 400 mg once per day, about 425 mg once per day, about 450 mg once per day, about 475 mg once per day, about 500 mg once per day, about 525 mg once per day, about 550 mg once per day, about 575 mg once per day, about 600 mg once per day, about 625 mg once per day, about 650 mg once per day, about 675 mg once per day, about 700 mg once per day, about 725 mg once per day, about 750 mg once per day, aboutWSGR Ref. 47134-784.601775 mg once per day, about 800 mg once per day, about 825 mg once per day, about 850 mg once per day, about 875 mg once per day, about 900 mg once per day, about 925 mg once per day, about 950 mg once per day, about 975 mg once per day, or about 1000 mg once per day.

[0143] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 100 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 200 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 300 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 400 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 500 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 600 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 700 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 800 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 900 mg once per day. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount of about 1000 mg once per day.

[0144] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 100 nM for at least 24 hours following administration.WSGR Ref. 47134-784.601

[0145] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 200 nM for at least 24 hours following administration.

[0146] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 250 nM for at least 24 hours following administration.

[0147] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to theWSGR Ref. 47134-784.601subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 275 nM for at least 24 hours following administration

[0148] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 300 nM for at least 24 hours following administration.

[0149] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nMWSGR Ref. 47134-784.601for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 400 nM for at least 24 hours following administration.

[0150] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 500 nM for at least 24 hours following administration.

[0151] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, whereinWSGR Ref. 47134-784.601Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 600 nM for at least 24 hours following administration.

[0152] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 625 nM for at least 24 hours following administration.

[0153] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 isWSGR Ref. 47134-784.601administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 650 nM for at least 24 hours following administration.

[0154] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 675 nM for at least 24 hours following administration.

[0155] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 700 nM for at least 24 hours following administration.

[0156] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma ofWSGR Ref. 47134-784.601the subject of equal to or greater than 725 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 725 nM for at least 24 hours following administration.

[0157] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 750 nM for at least 24 hours following administration.

[0158] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 8 hours following administration. In some embodiments are providedWSGR Ref. 47134-784.601the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 775 nM for at least 24 hours following administration.

[0159] In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 4 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 8 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 12 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 16 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 20 hours following administration. In some embodiments are provided the methods disclosed herein, wherein Compound 1 is administered to the subject in an amount that provides a concentration of Compound 1 in the plasma of the subject of equal to or greater than 800 nM for at least 24 hours following administration.

[0160] Furthermore, the pharmaceutically acceptable formulations of the compounds of Formula (I), or a pharmaceutically acceptable salt thereof, that may be used to practice the methods disclosed herein may contain a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in an amount of about 10 mg to about 2000 mg, or from about 10 mg to about 1500 mg, or from about 10 mg to about 1000 mg, or from about 10 mg to about 750 mg, or from about 10 mg to about 500 mg, or from about 25 mg to about 500 mg, or from about 50 mg to about 500 mg, or from about 100 mg to about 500 mg.

[0161] In some embodiments, the inhibitor of KRAS G12C and the EED inhibitor are administered to a subject in need thereof in 28-day cycles. In some embodiments, the inhibitor of KRAS G12C and the EED inhibitor are administered to a subject in need thereof in multiple 28-day cycles. In some embodiments, the inhibitor of KRAS G12C and the EED inhibitor are administered to a subject in needWSGR Ref. 47134-784.601thereof for at least one 28-day cycle. In some embodiments, the inhibitor of KRAS G12C and the EED inhibitor are administered to a subject in need thereof on each day of each 28-day cycle.

[0162] In some instances, the methods described herein comprise administering the inhibitor of KRAS G12C and the EED inhibitor to the subject or subject in need thereof in multiple cycles repeated on a regular schedule with periods of rest in between each cycle. For example, in some instances, treatment given for one week followed by three weeks of rest is one treatment cycle. The length of a treatment cycle depends on the treatment being given. In some embodiments, the length of a treatment cycle ranges from two to six weeks. In some embodiments, the length of a treatment cycle ranges from three to six weeks. In some embodiments, the length of a treatment cycle ranges from three to four weeks. In some embodiments, the length of a treatment cycle is three weeks (or 21 days). In some embodiments, the length of a treatment cycle is four weeks (28 days). In some embodiments, the length of a treatment cycle is 56 days. In some embodiments, a treatment cycle lasts one, two, three, or four weeks. In some embodiments, a treatment cycle lasts three weeks. In some embodiments, a treatment cycle lasts four weeks. The number of treatment doses scheduled within each cycle also varies depending on the drugs being given.

[0163] Dosages of compositions described herein can be determined by any suitable method. Maximum tolerated doses (MID) and maximum response doses (MRD) of the inhibitor of KRAS G12C and the EED inhibitor can be determined via established animal and human experimental protocols as well as in the examples described herein. For example, toxicity and therapeutic efficacy of the inhibitor of KRAS G12C and the EED inhibitor can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, for determining the LDso (the dose lethal to 50% of the population) and the EDso (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio between LDso and EDso. The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in a human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the EDso with minimal toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. Additional relative dosages, represented as a percent of maximal response or of maximum tolerated dose, are readily obtained via the protocols.

[0164] In some embodiments, the amount of the inhibitor of KRAS G12C and the EED inhibitor administered a subject in need thereof varies depending upon factors such as disease condition and its severity, the identity (e.g., age, weight, sex) of the subject or host in need of treatment, but can nevertheless be determined according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the liquid formulation type, the condition being treated, and the subject or host being treated.Methods of detecting biomarkers

[0165] The inhibitor of KRAS G12C administered to the subject having colorectal cancer may be used according to the methods disclosed herein or in an amount according to the information known to those ofWSGR Ref. 47134-784.601ordinary skill in the art regarding those agents. In one embodiment, the inhibitor of KRAS G12C is adagrasib and is orally administered to the subject having G12C mutated colorectal cancer in an amount that is about 600 mg twice daily. In another embodiment of the methods disclosed herein, adagrasib is orally administered to the subject in an amount that is about 600 mg twice daily, and Compound 1 is administered to the subject in an amount that is from about 200 mg to about 1000 mg once per day, or from about 200 mg to about 800 mg once per day, or from about 400 mg to about about 800 mg once per day. In some embodiments of the methods disclosed herein, adagrasib is orally administered to the subject in an amount that is about 600 mg twice daily, and Compound 1 is administered to the subject in an amount that is about 200 mg per day, or about 300 mg per day, or about 400 mg per day, or about 500 mg per day, or about 600 mg per day, or about 700 mg per day, or about 800 mg per day, or about 900 mg per day, or about 1000 mg per day.

[0166] In other embodiment, the inhibitor of KRAS G12C is sotorasib and is orally administered to the subject having G12C mutated colorectal cancer in an amount that is about 960 mg once daily. In another embodiment of the methods disclosed herein, sotorasib is orally administered to the subject in an amount that is about 960 mg once per day, and Compound 1 is administered to the subject in an amount that is from about 200 mg to about 1000 mg once per day, or from about 200 mg to about 800 mg once per day, or from about 400 mg to about about 800 mg once per day. In some embodiments of the methods disclosed herein, sotorasib is orally administered to the subject in an amount that is about 960 mg once per day, and Compound 1 is administered to the subject in an amount that is about 200 mg per day, or about 300 mg per day, or about 400 mg per day, or about 500 mg per day, or about 600 mg per day, or about 700 mg per day, or about 800 mg per day, or about 900 mg per day, or about 1000 mg per day.

[0167] In some embodiments are provided the methods disclosed herein wherein the measurement of one or more biomarkers is used to determine whether colorectal cancer in a subject comprises at KRAS G12C mutation and would benefit from the administration of an inhibitor of KRAS G12C, and an embryonic ectoderm development (EED) inhibitor. Methods of determining whether colorectal cancer in a subject comprises a KRAS G12C mutation are commercially available or are well known to those having ordinary skill in the art. For example, a biological sample from a subject having colorectal cancer (such as tumor tissue, blood, or plasma) is tested by methods known to those having ordinary skill in the art (such as by use of the QIAGEN therascreen KRAS RGQ PCR Kit) to determine whether the colorectal cancer in the subject comprises a KRAS G12C mutation prior to administration to the subject of (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor. If the colorectal cancer in the subject is determined to comprise a KRAS G12C mutation, then the subject is administered the inhibitor of KRAS G12C and the EED inhibitor according the methods disclosed herein.

[0168] Disclosed herein are methods comprising: (a) providing a biologic sample obtained from a subject having colorectal cancer; (b) assaying to detect in the biologic sample obtained from the subject a presence or absence of a biomarker, such as the presence of a KRAS G12C mutation; (c) detecting the presence or absence of the biomarker in the biologic sample using the methods described herein; and (d) administering to the subject a therapeutically effective amount of a compound of an inhibitor of KRASWSGR Ref. 47134-784.601G12C, and an embryonic ectoderm development (EED) inhibitor, if the biomarker is present in the biological sample.

[0169] The presence, absence, or level, of such biomarkers may be measured, collectively or individually, in a biological sample obtained from a subject, such as a sample of a solid tumor, such as colorectal cancer, or from a sample of a relevant biological fluid, such as a blood sample. In some instances, the one or more biomarkers are detected in plasma or serum that is derived from a blood sample obtained from the subject. In some instances, the methods of detection disclosed herein are useful for predicting a therapeutic response to a therapy described herein (e.g, the administration to a subj ect of an inhibitor of KRAS G12C, and an embryonic ectoderm development (EED) inhibitor), monitor the treatment using the therapy of, and treating with the therapy, a proliferative disease or condition described herein in a subject.

[0170] In some embodiments, the expression of a biomarker in a biological sample from a subject is measured by use of immunohistochemistry (IHC) assays. Such immunohistochemistry (IHC) assays are commercially available, or may be developed and utilized according to methods known to those having ordinary skill in the art.

[0171] Immunohistochemistry techniques utilize an antibody to probe and visualize cellular antigens in situ, generally by chromogenic or fluorescent methods. In such techniques, antibodies or antisera, polyclonal antisera, or monoclonal antibodies specific for each marker are used to detect expression. The antibodies can be detected by direct labeling of the antibodies themselves, for example, with radioactive labels, fluorescent labels, hapten labels such as, biotin, or an enzyme such as horse radish peroxidase or alkaline phosphatase. Alternatively, unlabeled primary antibody is used in conjunction with a labeled secondary antibody, comprising antisera, polyclonal antisera, or a monoclonal antibody specific for the primary antibody. Immunohistochemistry protocols and kits are well known in the art and are commercially available.

[0172] Two general methods of IHC are generally available; direct and indirect assays. According to the first assay, binding of antibody to the target antigen is determined directly. This direct assay uses a labeled reagent, such as a fluorescent tag or an enzyme-labeled primary antibody, which can be visualized without further antibody interaction. In a typical indirect assay, unconjugated primary antibody binds to the antigen and then a labeled secondary antibody binds to the primary antibody. Where the secondary antibody is conjugated to an enzymatic label, a chromagenic or Anorogenic substrate is added to provide visualization of the antigen. Signal amplification occurs because several secondary antibodies may react with different epitopes on the primary antibody. The primary and / or secondary antibody used for immunohistochemistry typically will be labeled with a detectable moiety. Numerous labels are available which can be generally grouped into the following categories. First, are radioisotopes, such as35S,14C,125I,3H, and131I. The antibody can be labeled with the radioisotope using the techniques described in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley -Interscience, New York, N.Y., Pubs. (1991) for example and radioactivity can be measured using scintillation counting. Next, are colloidal gold particles. Third are Auorescent labels including, but are not limited to, rare earth chelatesWSGR Ref. 47134-784.601(europium chelates), Texas Red, rhodamine, fluorescein, dansyl, Lissamine, umbelliferone, phycocrytherin, phycocyanin, or commercially available fluorophores such SPECTRUM ORANGE® and SPECTRUM GREEN® and / or derivatives of any one or more of the above. The fluorescent labels can be conjugated to the antibody using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a fluorimeter. Fourth are various enzyme- substrate labels are available and U.S. Pat. No. 4,275,149 provides a review of some of these. The enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (using a chemiluminometer, for example) or donates energy to a fluorescent acceptor. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3 -dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, f> -galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described in O'Sullivan et al. Methods for the Preparation of Enzyme- Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed J. Langone & H. Van Vunakis), Academic press, New York, 73:147-166 (1981). Examples of enzyme-substrate combinations include, for example (i) Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate, wherein the hydrogen peroxidase oxidizes a dye precursor [e.g., orthophenylene diamine (OPD) or 3,3 ’,5, 5 ’-tetramethyl benzidine hydrochloride (TMB)]. 3,3-Diaminobenzidine (DAB) may also be used to visualize the HRP-labeled antibody; (ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate as chromogenic substrate; and (iii) P-D-galactosidase ( -D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl-P-D-galactosidase) or Anorogenic substrate (e.g., 4-methylumbelliferyl-P-D-galactosidase). Numerous other enzyme- substrate combinations are available to those skilled in the art. These methods are generally described in U. S. Pat. Nos. 4,275,149 and 4,318,980. Sometimes, the label is indirectly conjugated with the antibody. The skilled artisan will be aware of various techniques for achieving this. For example, the antibody can be conjugated with biotin and any of the four broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the antibody in this indirect manner. Alternatively, to achieve indirect conjugation of the label with the antibody, the antibody is conjugated with a small hapten and one of the different types of labels mentioned above is conjugated with an anti-hapten antibody. Thus, indirect conjugation of the label with the antibody can be achieved.

[0173] Biological samples obtained from subjects comprising tissue samples may be prepared according to protocols commonly used in the art. Typically, sections of paraffin-embedded cells or tissues areWSGR Ref. 47134-784.601obtained by (1) preserving tissue in fixative, (2) dehydrating the fixed tissue, (3) infiltrating the tissue with fixative, (4) orienting the tissue such that the cut surface accurately represents the tissue, (5) embedding the tissue in paraffin (making a paraffin block), (6) cutting tissue paraffin block with a microtome in sections of 4-5 picometers, and (7) mounting sections onto slides. The slides may then be read by a pathologist or the like assessing for the presence or absence of a biomarker, or of abnormal or normal cells or a specific cell type and provides the loci of the cell types of interest. Thus, for example, a pathologist or the like would review the slides and identify normal cells and abnormal cells (such as abnormal or tumor cells). Any means of defining the loci of the cells of interest may be used (e.g., coordinates on an X-Y axis).

[0174] Aside from the sample preparation procedures discussed above, further treatment of the tissue section prior to, during or following IHC may be desired. For example, epitope retrieval methods, such as heating the tissue sample in citrate buffer may be carried out [see, e.g., Leong et al. Appl.Immunohistochem. 4(3):201 (1996)]. Following an optional blocking step, the tissue section is exposed to primary antibody for a sufficient period of time and under suitable conditions such that the primary antibody binds to the target protein antigen in the tissue sample. Appropriate conditions for achieving this can be determined by routine experimentation.

[0175] The extent of binding of antibody to the sample is determined by using any one of the detectable labels discussed above. For example, the label is an enzymatic label (e.g. HRPO) which catalyzes a chemical alteration of the chromogenic substrate such as 3, 3 ’-diaminobenzidine chromogen. Preferably the enzymatic label is conjugated to antibody which binds specifically to the primary antibody (e.g. the primary antibody is rabbit polyclonal antibody and secondary antibody is goat anti-rabbit antibody). Specimens thus prepared may be mounted and coverslipped. Slide evaluation is then determined, e.g. using a microscope.

[0176] IHC may be combined with morphological staining, either prior to or thereafter. After deparaffinization, the sections mounted on slides may be stained with a morphological stain for evaluation. The morphological stain to be used provides for accurate morphological evaluation of a tissue section. The section may be stained with one or more dyes each of which distinctly stains different cellular components. In one embodiment, hematoxylin is use for staining cellular nucleic of the slides. Hematoxylin is widely available. An example of a suitable hematoxylin is Hematoxylin II (Ventana). When lighter blue nuclei are desired, a bluing reagent may be used following hematoxylin staining. One of skill in the art will appreciate that staining may be optimized for a given tissue by increasing or decreasing the length of time the slides remain in the dye.

[0177] Automated systems for slide preparation and IHC processing are available commercially. The Ventana® BenchMark XT system is an example of such an automated system.

[0178] After staining, the tissue section may be analyzed by standard techniques of microscopy.Generally, a pathologist or the like assesses the tissue for the presence of abnormal or normal cells or a specific cell type and provides the loci of the cell types of interest. Thus, for example, a pathologist or the like would review the slides and identify normal cells and abnormal cells (such as abnormal or tumorWSGR Ref. 47134-784.601cells). Any means of defining the loci of the cells of interest may be used (e.g., coordinates on an X-Y axis).

[0179] In some embodiments, the presence, or an absence, and / or a level of expression of the biomarker is detected in the sample obtained from a subject by analyzing the genetic material in the sample. In some embodiments, the genetic material is obtained from blood, serum, plasma, sweat, hair, tears, urine, and other techniques known by one of skill in the art. In some embodiments the sample comprises circulating tumor RNA (ctRNA). In some embodiments the sample comprises peripheral blood mononuclear cells (PBMCs). In some embodiments the sample comprises circulating tumor cells (CTCs). In some cases, the genetic material is obtained from a tumor biopsy or liquid biopsy. In some embodiments, a tumor biopsy comprises a formalin-fixed paraffin embedded biopsy, a fresh frozen biopsy, a fresh biopsy, or a frozen biopsy. In some embodiments, a liquid biopsy comprises PBMCs, circulating tumor RNA, plasma cell-free RNA, or circulating tumor cells (CTCs). Tumor and liquid biopsies can undergo additional analytic processing for sample dissociation, cell sorting, and enrichment of cell populations of interest.

[0180] In some embodiments, methods of detecting a presence, absence, or level of a biomarker in a biologic sample obtained from the subject involve detecting a nucleic acid sequence. In some cases, the nucleic acid sequence comprises deoxyribonucleic acid (DNA), such as in the case of detecting complementary DNA (cDNA) of an mRNA transcript. In some instances, the nucleic acid sequence comprises a denatured DNA molecule or fragment thereof. In some instances, the nucleic acid sequence comprises DNA selected from: genomic DNA, viral DNA, mitochondrial DNA, plasmid DNA, amplified DNA, circular DNA, circulating DNA, cell -free DNA, or exosomal DNA. In some instances, the DNA is single-stranded DNA (ssDNA), double-stranded DNA, denaturing double- stranded DNA, synthetic DNA, and combinations thereof. The circular DNA may be cleaved or fragmented. In some instances, the nucleic acid sequence comprises ribonucleic acid (RNA). In some instances, the nucleic acid sequence comprises fragmented RNA. In some instances, the nucleic acid sequence comprises partially degraded RNA. In some instances, the nucleic acid sequence comprises a microRNA or portion thereof. In some instances, the nucleic acid sequence comprises an RNA molecule or a fragmented RNA molecule (RNA fragments) selected from: a microRNA (miRNA), apre-miRNA, apri-miRNA, a mRNA, apre-mRNA, a viral RNA, a viroid RNA, a virusoid RNA, circular RNA (circRNA), a ribosomal RNA (rRNA), a transfer RNA (tRNA), apre-tRNA, a long non-coding RNA (IncRNA), a small nuclear RNA (snRNA), a circulating RNA, a cell-free RNA, an exosomal RNA, a vector-expressed RNA, an RNA transcript, a synthetic RNA, and combinations thereof.

[0181] Disclosed herein, in some embodiments, a biomarker is detected by subjecting a sample obtained from the subject to a nucleic acid-based detection assay. In some instances, the nucleic acid-based detection assay comprises quantitative polymerase chain reaction (qPCR), reverse transcription PCT (RT-qPCR), gel electrophoresis (including for e.g., Northern or Southern blot), immunohistochemistry (IHC), immunofluorescence (IF), in situ hybridization (ISH) such as fluorescent in situ hybridization (FISH), cytochemistry, microarray, or sequencing. In some embodiments, the sequencing technique comprises next generation sequencing. In some embodiments, the methods involve a hybridization assay such asWSGR Ref. 47134-784.601fluorogenic qPCR(e.g., TaqMan™, SYBR green, SYBR green I, SYBR green II, SYBR gold, ethidium bromide, methylene blue, Pyronin Y, DAPI, acridine orange, Blue View or phycoerythrin), which involves a nucleic acid amplification reaction with a specific primer pair, and hybridization of the amplified nucleic acid probes comprising a detectable moiety or molecule that is specific to a target nucleic acid sequence. In some instances, a number of amplification cycles for detecting a target nucleic acid in a qPCR assay is about 5 to about 30 cycles. In some instances, the number of amplification cycles for detecting a target nucleic acid is at least about 5 cycles. In some instances, the number of amplification cycles for detecting a target nucleic acid is at most about 30 cycles. In some instances, the number of amplification cycles for detecting a target nucleic acid is about 5 to about 10, about 5 to about 15, about 5 to about 20, about 5 to about 25, about 5 to about 30, about 10 to about 15, about 10 to about 20, about 10 to about 25, about 10 to about 30, about 15 to about 20, about 15 to about 25, about 15 to about 30, about 20 to about 25, about 20 to about 30, or about 25 to about 30 cycles. For TaqMan™ methods, the probe may be a hydrolysable probe comprising a fluorophore and quencher that is hydrolyzed by DNA polymerase when hybridized to a target nucleic acid. In some cases, the presence of a target nucleic acid is determined when the number of amplification cycles to reach a threshold value is less than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, or 20 cycles. In some instances, hybridization may occur at standard hybridization temperatures, e.g. , between about 35 °C and about 65 °C in a standard PCR buffer.

[0182] An additional exemplary nucleic acid-based detection assay comprises the use of nucleic acid probes conjugated or otherwise immobilized on a bead, multi -well plate, or other substrate, wherein the nucleic acid probes are configured to hybridize with a target nucleic acid sequence. In some instances, the nucleic acid probe is specific to one or more of a polynucleotide sequence that encodes a relevant biomarker as disclosed herein. In some instances, the nucleic acid probe specific to a biomarker comprises a nucleic acid probe sequence sufficiently complementary to the polynucleotide sequence that encodes the relevant biomarker protein. In some instances, the probe comprises a transcribed polynucleotide sequence (e.g., RNA, cDNA). In some embodiments, the nucleic acid probe can be, for example, a full-length cDNA, or a portion thereof, such as an oligonucleotide of at least about 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides in length and sufficient to specifically hybridize under standard hybridization conditions to the target nucleic acid sequence. In some embodiments, the target nucleic acid sequence is immobilized on a solid surface and contacted with a probe, for example by running the isolated target nucleic acid sequence on an agarose gel and transferring the target nucleic acid sequence from the gel to a membrane, such as nitrocellulose. In some embodiments, the probe(s) are immobilized on a solid surface, for example, in an Affymetrix gene chip array, and the probe(s) are contacted with the target nucleic acid sequence.

[0183] In some embodiments, the term “probe” with regards to nucleic acids, refers to any nucleic acid molecule that is capable of selectively binding to a specifically intended target nucleic acid sequence. In some instances, probes are specifically designed to be labeled, for example, with a radioactive label, a fluorescent label, an enzyme, a chemiluminescent tag, a colorimetric tag, or other labels or tags that are known in the art. In some instances, the fluorescent label comprises a fluorophore. In some instances, theWSGR Ref. 47134-784.601fluorophore is an aromatic or heteroaromatic compound. In some instances, the fluorophore is a pyrene, anthracene, naphthalene, acridine, stilbene, benzoxazole, indole, benzindole, oxazole, thiazole, benzothiazole, canine, carbocyanine, salicylate, anthranilate, xanthenes dye, coumarin. Exemplary xanthene dyes include, e.g., fluorescein and rhodamine dyes. Fluorescein and rhodamine dyes include, but are not limited to 6- carboxy fluorescein (FAM), 2'7'-dimethoxy-4'5'-dichloro-6-carboxyfluorescein (JOE), tetrachlorofluorescein (TET), 6-carboxyrhodamine (R6G), N,N,N; N'-tetramethyl-6-carboxyrhodamine (TAMRA), 6-carboxy-X-rhodamine (ROX). Suitable fluorescent probes also include the naphthylamine dyes that have an amino group in the alpha or beta position. For example, naphthylamino compounds include l-dimethylaminonaphthyl-5-sulfonate, l-anilino-8-naphthalene sulfonate, and 2-p-toluidinyl-6-naphthalene sulfonate, 5-(2'-aminoethyl)aminonaphthalene-l-sulfonic acid (EDANS). Exemplary coumarins include, e.g., 3-phenyl-7-isocyanatocoumarin; acridines, such as 9-isothiocyanatoacridine and acridine orange; N-(p-(2-benzoxazolyl)phenyl) mal eimide; cyanines, such as, e.g., indodi carbocyanine 3 (Cy3), indodicarbocyanine 5 (Cy5), indodicarbocyanine 5.5 (Cy5.5), 3-(-carboxy-pentyl)-3'-ethyl-5,5'-dimethyloxacarbocyanine (CyA); 1H, 5H, 11H, 15H-Xantheno[2,3, 4-ij: 5,6, 7-i'j'] di quinolizin- 18-ium, 9-[2 (or 4)-[[[6-[2,5-dioxo-l-pyrrolidinyl)oxy]-6-oxohexyl]amino]sulfonyl]-4 (or 2)-sulfophenyl]-2,3, 6,7, 12,13, 16,17-octahydro-inner salt (TRor Texas Red); or BODIPYTM dyes. In some cases, the probe comprises FAM as the dye label.

[0184] In some embodiments, detecting the one or more biomarkers comprises sequencing genetic material obtained from a sample from the subject. Sequencing can be performed with any appropriate sequencing technology, including but not limited to single-molecule real-time (SMRT) sequencing, Polony sequencing, sequencing by ligation, reversible terminator sequencing, proton detection sequencing, ion semiconductor sequencing, nanopore sequencing, electronic sequencing, pyrosequencing, Maxam-Gilbert sequencing, chain termination (e.g., Sanger) sequencing, +S sequencing, or sequencing by synthesis. Sequencing methods also include next-generation sequencing, e.g., modern sequencing technologies such as Illumina sequencing (e.g., Solexa), Roche 454 sequencing, Ion torrent sequencing, and SOLiD sequencing. In some cases, next-generation sequencing involves high-throughput sequencing methods. Additional sequencing methods available to one of skill in the art may also be employed.

[0185] In some instances, a number of nucleotides that are sequenced are at least 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 300, 400, 500, 2000, 4000, 6000, 8000, 10000, 20000, 50000, 100000, or more than 100000 nucleotides. In some instances, the number of nucleotides sequenced is in a range of about 1 to about 100000 nucleotides, about 1 to about 10000 nucleotides, about 1 to about 1000 nucleotides, about 1 to about 500 nucleotides, about 1 to about 300 nucleotides, about 1 to about 200 nucleotides, about 1 to about 100 nucleotides, about 5 to about 100000 nucleotides, about 5 to about 10000 nucleotides, about 5 to about 1000 nucleotides, about 5 to about 500 nucleotides, about 5 to about 300 nucleotides, about 5 to about 200 nucleotides, about 5 to about 100 nucleotides, about 10 to about 100000 nucleotides, about 10 to about 10000 nucleotides, about 10 to about 1000 nucleotides, about 10 to about 500 nucleotides, about 10 to about 300 nucleotides, about 10 to about 200 nucleotides, about 10 to about 100 nucleotides, about 20 to about 100000 nucleotides, about 20 to about 10000 nucleotides, aboutWSGR Ref. 47134-784.60120 to about 1000 nucleotides, about 20 to about 500 nucleotides, about 20 to about 300 nucleotides, about 20 to about 200 nucleotides, about 20 to about 100 nucleotides, about 30 to about 100000 nucleotides, about 30 to about 10000 nucleotides, about 30 to about 1000 nucleotides, about 30 to about 500 nucleotides, about 30 to about 300 nucleotides, about 30 to about 200 nucleotides, about 30 to about 100 nucleotides, about 50 to about 100000 nucleotides, about 50 to about 10000 nucleotides, about 50 to about 1000 nucleotides, about 50 to about 500 nucleotides, about 50 to about 300 nucleotides, about 50 to about 200 nucleotides, or about 50 to about 100 nucleotides.

[0186] In some cases, a hybridization assay, such as those described herein, is used to detect the mRNA encoding the biomarker in the sample. Exemplary probe sequences that are hybridizable to a target nucleic acid sequence comprise at least 10, but no more than 100 contiguous nucleotides comprising the relevant sequence. In some cases, RNA sequencing (RNAseq) is used to detect the mRNA encoding the relevant biomarker protein.

[0187] Detection of the mRNA, in some cases, involves amplification of the subject’s nucleic acid by the polymerase chain reaction (PCR). In some embodiments, the PCR assay involves use of a pair of primers capable of amplifying at least about 10 contiguous nucleobases within a nucleic acid sequence, thereby amplifying the one or more gene products in the biomarker. In Anorogenic quantitative PCR, quantitation is based on amount of Auorescence signals (TaqMan and SYBR green). In some embodiments, the nucleic acid probe is conjugated to a detectable molecule. The detectable molecule may be a Auorophore. The nucleic acid probe may also be conjugated to a quencher.

[0188] In some embodiments, the assay for detecting the presence or absence of mRNA encoding a relevant biomarker comprises reverse-transcribing the relevant mRNA molecule to produce a corresponding complementary DNA (cDNA) molecule. In some embodiments, the assay further comprises contacting the cDNA molecule with a nucleic acid probe comprising a nucleic acid sequence that is complementary to a nucleic acid sequence of the cDNA molecule. In some embodiments, the assay comprises detecting a double-stranded hybridization product between the nucleic acid probe and the cDNA molecule. In some embodiments, the hybridization product is further ampli tied using a pair of primers. In some embodiments, the primers comprises a first primer with a nucleic acid sequence comprising at least 10 but not more than 50 contiguous nucleic acids within a relevant nucleic acid sequence that binds to a top strand of the double- stranded hybridization product; and a second primer with a nucleic acid sequence comprising at least 10 but not more than 50 contiguous nucleic acids within a nucleic acid sequence that is reverse complement to the relevant nucleic acid sequence that binds to a bottom strand of the double-stranded hybridization product.

[0189] Disclosed herein, in some embodiments, are methods comprising preparing a complementary DNA (cDNA) library. In some embodiments, the cDNA library is sequenced using suitable sequence methodologies disclosed herein. In some embodiments, the cDNA library is labeled, a plurality of nucleic acid probes is generated, and fixed to an immobile surface (such as a microarray). In some embodiments, the plurality of nucleic acid probes is capable of hybridizing to at least about 10 contiguous nucleotides of the two or more genes in a sample obtained from the subject. In some embodiments, detecting theWSGR Ref. 47134-784.601presence of or absence of a biomarker includes detecting a high or a low level of expression of one or more genes as compared to a reference level.

[0190] Disclosed herein, in some embodiments, genetic material is extracted from a biologic sample obtained from a subject, e.g., a sample of blood, serum, or tissue. In certain embodiments where nucleic acids are extracted, the nucleic acids are extracted using any technique that does not interfere with subsequent analysis. In certain embodiments, this technique uses alcohol precipitation using ethanol, methanol, or isopropyl alcohol. In certain embodiments, this technique uses phenol, chloroform, or any combination thereof. In certain embodiments, this technique uses cesium chloride. In certain embodiments, this technique uses sodium, potassium or ammonium acetate or any other salt commonly used to precipitate DNA. In certain embodiments, this technique utilizes a column or resin based nucleic acid purification scheme such as those commonly sold commercially, one non-limiting example would be the GenElute Bacterial Genomic DNA Kit available from Sigma Aldrich. In certain embodiments, after extraction the nucleic acid is stored in water, Tris buffer, or Tris-EDTA buffer before subsequent analysis. In an exemplary embodiment, the nucleic acid material is extracted in water. In some cases, extraction does not comprise nucleic acid purification. In certain embodiments, RNA may be extracted from cells using RNA extraction techniques including, for example, using acid phenol / guanidine isothiocyanate extraction (RNAzol B; Biogenesis), RNeasy RNA preparation kits (Qiagen) or PAXgene (PreAnalytix, Switzerland).

[0191] In some aspects, circulating tumor RNA (ctRNA) is used to assess the expression levels of RNA molecules, shed by the tumor into the blood stream. In some embodiments, detection of ctRNA is useful, for example, for detecting and diagnosing a tumor. Because tumor DNA and RNA has acquired multiple genetic mutations, leading to tumor development, ctRNA are not an exact match to the individual’s DNA and RNA, respectively. Finding DNA and RNA with genetic differences aids in tumor detection.Diagnosing the type of tumor using ctRNA can reduce the need for getting a sample of the tumor tissue (tumor biopsy), which can be challenging when a tumor is difficult to access, such as a tumor in the brain or lung.

[0192] In some embodiments, a decrease in the quantity of ctRNA suggests the solid tumor is shrinking and treatment with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor is effective. In some embodiments, a lack of ctRNA in the bloodstream indicates that the cancer has not returned after treatment with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor.

[0193] Described herein are methods of assessing genetic alterations by ctRNA profiling. In some embodiments, the genomic profiling is performed after each treatment cycle with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor. In some embodiments, the gene alterations indicate that the cancer is becoming resistant to the treatment with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor. In some embodiments, the lack of gene alterations indicate that the cancer is not becoming resistant to the treatment with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor.WSGR Ref. 47134-784.601

[0194] Described herein are methods of assessing genetic alterations, including, but not limited to, mutations in certain genes and / or copy number alterations in certain genes, by circulating tumor DNA (ctDNA) and / or cell -free DNA (cfDNA) profiling. In some embodiments, the genomic profiling is performed after each treatment cycle with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor. In some embodiments, the gene alterations indicate that the cancer is becoming resistant to the treatment with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor. In some embodiments, the lack of gene alterations indicate that the cancer is not becoming resistant to the treatment with an inhibitor of KRAS G12C and an embryonic ectoderm development (EED) inhibitor.

[0195] In some embodiments, the expression of a biomarker is measured by immunofluorescence (IF) assays. In some embodiments, the expression of a biomarker is measured by in situ hybridization (ISH) assays. In some embodiments, the expression of a biomarker transcript levels are measured using assays such as quantitative polymerase chain reaction (qPCR), microarray, and RNA sequencing, or assays commercially available from companies such as Fluidigm and Nanostring.

[0196] Disclosed herein are methods of treating a subject having colorectal cancer, comprising: (a) providing a biologic sample obtained from a subject having colorectal cancer; (b) assaying to detect in the biologic sample obtained from the subject a presence or absence of a biomarker, such as a KRAS G12C mutation; (c) detecting the presence or absence of the biomarker in the biologic sample using the methods described herein; and (d) administering to the subject an inhibitor of KRAS G12C, and an embryonic ectoderm development (EED) inhibitor, if the biomarker is present in the biological sample. In some embodiments, expression of a biomarker is based on the expression level of the biomarker deviating from a reference expression level. In some embodiments, the expression level is high, relative to the reference expression level. In some embodiments, the expression level is low, relative to the reference expression level. In some embodiments, the reference expression level is derived from an individual, or a group of individuals, that do not have cancer. In some embodiments, the reference expression level is derived from an individual, or a group of individuals, that have cancer that does not therapeutically respond to the inhibitor of KRAS G12C and the EED inhibitor. In some embodiments, the expression level deviates from the reference expression level by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.

[0197] In some embodiments, the determination of expression or the presence of a biomarker is defined based on the percentage of cells that stain weakly, moderately, or strongly for the relevant biomarker, with the threshold defining the minimal percentage of cells that are required to stain positive at the various intensity levels (>a% of tumor cells stain weakly, >b% of tumor cells stain moderately, >c% of tumor cells stain strongly, or a combination thereof). In some embodiments, the one or more of the cells comprising the cancer has been determined to express a biomarker when > about 10%, > about 15%, > about 20%, > about 25%, > about 30%, > about 35%, > about 40%, > about 45%, > about 50%, > about 55%, > about 60%, > about 65%, > about 70%, > about 75%, > about 80%, > about 85%, > about 90%, or > about 95% of the tumor cells stain weakly for the biomarker; when > about 10%, > about 15%, > about 20%, > about 25%, > about 30%, > about 35%, > about 40%, > about 45%, > about 50%, > about 55%, >WSGR Ref. 47134-784.601about 60%, > about 65%, > about 70%, > about 75%, > about 80%, > about 85%, > about 90%, or > about 95% of the tumor cells stain moderately for the biomarker; when > about 10%, > about 15%, > about 20%, > about 25%, > about 30%, > about 35%, > about 40%, > about 45%, > about 50%, > about 55%, > about 60%, > about 65%, > about 70%, > about 75%, > about 80%, > about 85%, > about 90%, or > about 95% of the tumor cells stain strongly for the biomarker; or any combinations thereof.Kits and Articles of Manufacture

[0198] Disclosed herein, in certain embodiments, are kits and articles of manufacture for use with one or more methods and compositions described herein. Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In one embodiment, the containers are formed from a variety of materials such as glass or plastic.

[0199] A kit typically includes labels listing contents and / or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.

[0200] In one embodiment, a label is on or associated with the container. In one embodiment, a label is on a container when letters, numbers or other characters forming the label are attached, molded, or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In one embodiment, a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein.

[0201] In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. In one embodiment, the pack or dispenser device is accompanied by instructions for administration. In one embodiment, the pack or dispenser is also 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, is the labeling approved by the U.S. Food and Drug Administration for drugs, or the approved product insert. In one embodiment, compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

[0202] Disclosed herein is a kit comprising (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, for use in treating colorectal cancer in a subject in need thereof and a package insert comprising instructions for administering to the subject having colorectal cancer the inhibitor of KRAS G12C and the EED inhibitor using if the colorectal cancer in the subject is determined to comprise a KRAS G12C mutation.Numbered EmbodimentsWSGR Ref. 47134-784.601

[0203] Embodiment 1. A method of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation.

[0204] Embodiment 2. A method of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation by an FDA-approved test.

[0205] Embodiment 3. A method of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0206] Embodiment 4. A method of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation by an FDA approved test, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0207] Embodiment 5. A method of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor.

[0208] Embodiment 6. The method of any one of Embodiments 1 to 5, wherein the subject has not acquired one or more secondary site pathway mutations.

[0209] Embodiment 7. The method of Embodiment 6, wherein the secondary site pathway mutation is a RAS / MAPK pathway alteration, optionally a KRAS alternation,a KRAS-activating mutation, optionally a mutation in codon 12 or 13,a KRAS amplification,a RAF / MAPK pathway mutation or fusion, optionally MAP2K1K57and BRAFV600E, a KRAS switch-II pocket mutation, optionally KRAS Y96D,an NRAS / HRAS mutation, optionally a single nucleotide variant in codon 61,a RTK amplification, capable of re-activating or mediating reactivation of a RAS / MAPK signaling pathway, optionally wherein the RTK amplification is MET amplification, ERBB2 amplification, ERBB4 amplification, EGFR amplification, or FGFR2 amplification, ora RTK fusion, capable of re-activating or mediating reactivation of a RAS / MAPK signaling pathway, optionally wherein the RTK fusion is CCDC6-RET fusion, EML4-ALK fusion, FGFR3-TACC fusion, AHCYL2-MET fusion, MET-SLC12A9 fusion, or GOPC-ROS1 fusion, orany combination thereof.

[0210] Embodiment 8. The method of Embodiment 7, wherein:the KRAS-activating mutation is KRAS G12A, KRAS G12D, KRAS G12V, KRAS G13D, or KRAS Q61H;the KRAS switch-II pocket mutation is KRAS Y64 mutation, KRAS H95 mutation, KRAS Y96 mutation, optionally R68S, M72T, H95R, H95N, H95L, H95Q, Y96D, Y96N, Y96H, or Q99L; orWSGR Ref. 47134-784.601the NRAS mutation is NRAS Q61K, or NRAS Q61R.

[0211] Embodiment 9. The method of any one of Embodiments 1 to 8, wherein the subject has received one or more chemotherapeutic treatments prior to the administration to the subject of the inhibitor of KRAS G12C and the EED inhibitor, optionally wherein the one or more chemotherapeutic treatments are selected from fluoropyrimidine-based chemotherapy, oxaliplatin-based chemotherapy, and irinotecanbased chemotherapy.

[0212] Embodiment 10. The method of any one of Embodiments 1 to 9, wherein the subject is further administered one or more anti-EGFR agents, optionally wherein the one or more anti-EGFR agents are selected from EGFR antagonists, inhibitors, degraders, and antibody-drug conjugates.

[0213] Embodiment 11. The method of Embodiment 10, wherein the one or more anti-EGFR agents are selected from panitumumab, cetuximab, osimertinib, erlotinib, gefitinib, lazertinib, firmonertinib, sunvozertinib, zipalertinib, ORIC-114, silevertinib (BDTX-1535), zanidatamab (ZW25), nimotuzumab, cetuximab saratolacan, depatuxizumab mafodotin, telisotuzumab vedotin (ABBV-399), AVID100, C225-ILs-dox, MRG003, E-EDV-D682, EGFR(V)-EDV-dox, EGR-ErbituxEDVsMIT, laptrituximab emtansine, losatuxizumab vedotin, serclutamab talirine, LD-DM1, R68-MC-VC-PAB-MMAE, RC68-PY-VC-PAB-MMAE, SHR-A1307, cetuximab-triptolide, M1231, and B2C4-MMAE.

[0214] Embodiment 12. The method of any one of Embodiments 1 to 11, wherein the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1 AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400 RMC-4998, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, andYL-17231 (TEB-17231).

[0215] Embodiment 13. The method of Embodiment 12, wherein the inhibitor of KRAS G12C is adagrasib or sotorasib.

[0216] Embodiment 14. The method of any one of Embodiments 1 to 13, wherein the EED inhibitor is a small molecule having a molecular weight of less or equal to 3000 Daltons.

[0217] Embodiment 15. The method of any one of Embodiments 1 to 14, wherein the EED inhibitor is selected from EED226, A-395, APG-5918, BR-001, BR-002, EEDi-5285, EEDi-1056, FTX-6274, pociredir (FTX-6058), HJM-353, and MAK683.

[0218] Embodiment 16. The method of any one of Embodiments 1 to 14, wherein the EED inhibitor is a compound of Formula (I)WSGR Ref. 47134-784.601XRM zFormula (I)or a pharmaceutically acceptable salt thereof:wherein:- represents a single or a double bond;Z is O or S;X is O, CR5, CR5OH, or C(R5)2, wherein:when X is O, - is a single bond;when X is C(R5)2, - is a single bond;when X is CR5OH, - is a single bond; orwhen X is CR5, - is a double bond;R1is aryl, heteroaryl, L-cycloalkyl, -N(R5)heterocyclyl, or L-heterocyclyl, wherein the aryl, the heteroaryl or the cyclyl portion of the L-cycloalkyl, -N(R5)heterocyclyl, or L-heterocyclyl is optionally substituted with one or more R4;R2is cyano, -COOR5, -C(O)N(R5)2, or -C(O)N(R5)2 wherein each R5taken together with the nitrogen atom to which they are attached form a 5 - 8 membered heterocyclic ring optionally substituted with one or more R4;each R3is independently C1-C3 alkyl or halogen;each R4is independently oxo, cyano, halogen, -PCLCCi-Cs alkyl)2, hydroxyl, alkoxy, hydroxyalkyl, heteroalkyl, aralkyl, haloalkyl, -COOR5, -Y2-haloalkyl, -Y^Ci-Ce alkyl, -Y2-Ci-Ce alkyl, -L-cycloalkyl, -L-heteroaryl, -L-heterocyclyl, -Y'-heterocyclyl. -Y2-heterocyclyl, -L- N(R5)2, -O-L-N(R5)2, -C(CF3)N(R5)2, -Y1-N(R5)2, or -Y2-N(R5)2, wherein the ring portion of the aralkyl, -L-cycloalkyl, -L-heteroaryl, -L-heterocyclyl or -Y'-heterocycl l is optionally substituted with one or more R7;L is a bond or C1-C4 alkylene;Y1is a bond, -C(O)-, or -NHC(O)-;Y2is a bond, -S-, -SO-, -SO2-, or -NR5SO2-,each R5is hydrogen or C1-C3 alkyl;R6is hydrogen, C1-C3 alkyl, halogen, haloalkyl, hydroxyalkyl, or heteroalkyl;each R7is oxo, cyano, hydroxyl, alkoxy, halogen, haloalkyl, hydroxyalkyl, heteroalkyl, cycloalkyl, -L-N(R5)2, Ci-Ce alkyl, or -Y'-heterocyclyl; andn is 1 or 2.WSGR Ref. 47134-784.601

[0219] Embodiment 17. The method of Embodiment 16, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, Z is O.

[0220] Embodiment 18. The method of Embodiment 17, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, Z is S.

[0221] Embodiment 19. The method of any one of Embodiments 16 to 18, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, n is 1.

[0222] Embodiment 20. The method of any one of Embodiments 16 to 19, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R2is cyano.

[0223] Embodiment 21. The method of any one of Embodiments 16 to 19, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R2is -COOR5.

[0224] Embodiment 22. The method of any one of Embodiments 16 to 19, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R2is -C(O)N(R5)2.

[0225] Embodiment 23. The method of any one of Embodiments 16 to 22, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R3is halogen.

[0226] Embodiment 24. The method of Embodiment 23, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R3is fluorine.

[0227] Embodiment 25. The method of any one of Embodiments 16 to 24, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, X is C(R5)2 and - is a single bond.

[0228] Embodiment 26. The method of any one of Embodiments 16 to 24, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, X is CR5and - is a double bond.

[0229] Embodiment 27. The method of any one of Embodiments 16 to 24, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, X is O and - is a single bond.

[0230] Embodiment 28. The method of any one of Embodiments 16 to 27, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is aryl optionally substituted with one or more R4.

[0231] Embodiment 29. The method of Embodiment 28, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is phenyl optionally substituted with one or more R4.

[0232] Embodiment 30. The method of Embodiment 29, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is phenyl is substituted with one, two or three R4.

[0233] Embodiment 31. The method of Embodiment 30, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, the one, two or three R4are each independently halogen, -PO3(Ci-C3 alkyl)2, hydroxyl, hydroxyalkyl, aralkyl, haloalkyl, -COOR5, -Y^Ci-Ce alkyl, Y2-Ci-Ce alkyl, -L-N(R5)2, -O-L-N(R5)2, -C(CF3)N(R5)2, -Y1-N(R5)2, -Y2-N(R5)2, Y2-haloalkyl, -L-heteroaryl, -L-heterocyclyl, or -Y'-heterocyclyl. wherein the heterocyclyl portion of the -L-heterocyclyl or -Y1-heterocyclyl is optionally substituted with one or more R7.

[0234] Embodiment 32. The method of Embodiment 31, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y^Ci-Ce alkyl and Y1is a bond and the Ci-Ce alkyl is methyl, ethyl, isopropyl, butyl, or pentyl.WSGR Ref. 47134-784.601

[0235] Embodiment 33. The method of Embodiment 31, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y2-CI-C6 alkyl and Y2is a -SO2- and the Ci-Ce alkyl is methyl.

[0236] Embodiment 34. The method of Embodiment 31, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y2-haloalkyl and Y2is -S- or -SO2- and the haloalkyl is trifluoromethyl.

[0237] Embodiment 35. The method of Embodiment 31, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-N(R5)2 and L is a bond and each R5is hydrogen, each R5is methyl, or one R5is methyl and one R5is hydrogen.

[0238] Embodiment 36. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-N(R5)2 and L is methylene or ethylene and each R5is hydrogen, each R5is methyl or one R5is methyl and one R5is hydrogen.

[0239] Embodiment 37. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y1-N(R5)2, Y1is -C(O)- and each R5independently is hydrogen, each R5is independently methyl or one R5is methyl and one R5is hydrogen.

[0240] Embodiment 38. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y2-N(R5)2, Y2is -SO2- and each R5independently is hydrogen, each R5is methyl, or one R5is methyl and one R5is independently hydrogen.

[0241] Embodiment 39. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y1-heterocyclyl and Y1is -C(O)- and the heterocyclyl portion of the L-heterocyclyl is piperazinyl or 4-methyl-piperazinyl.

[0242] Embodiment 40. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-heterocyclyl and L is a bond and the heterocyclyl portion of the L-heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or 3-azabicyclo[3.1.0]hexanyl, each optionally substituted with one or more R7selected from oxo, C1-C3 alkyl, alkoxy, hydroxyl and halogen.

[0243] Embodiment 41. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-heterocyclyl, wherein L is a methylene and the heterocyclyl portion of the L-heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl piperidinyl, each optionally substituted with one or more R7selected from C1-C3 alkyl, alkoxy, hydroxyl and halogen.

[0244] Embodiment 42. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y1-heterocyclyl and Y1is -C(O)- and the heterocyclyl portion of the Y1-heterocyclyl is morpholinyl optionally substituted with one or more C1-C3 alkyl.

[0245] Embodiment 43. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-heteroaryl optionally substituted with one or more R7.

[0246] Embodiment 44. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is tetrazolyl.WSGR Ref. 47134-784.601

[0247] Embodiment 45. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -PO3(Ci-C3 alkyl)2.

[0248] Embodiment 46. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -COOR5.

[0249] Embodiment 47. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is hydroxyalkyl.

[0250] Embodiment 48. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -O-L-N(R5)2.

[0251] Embodiment 49. The method of Embodiment 31 , wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is aralkyl.

[0252] Embodiment 50. The method of any one of Embodiments 16 to 27, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is heteroaryl optionally substituted with one or more R4.

[0253] Embodiment 51. The method of Embodiment 50, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazinyl, pyridyl, pyridinyl -2-one, pyrazinyl, pyridazinyl, pyrimidinyl, isoxazolyl, isoindolinyl, naphthyridinyl, 1,2,3,4-tetrahydroisoquinolinyl, or 5,6-dihydro-4H-pyrrolo[l,2-b]pyrazolyl, each optionally substituted with one or more R4.

[0254] Embodiment 52. The method of Embodiment 50 or 51, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is substituted with one or more R4; wherein each R4is independently cyano, halogen, -Y^Ci-Ce alkyl, -Y2-CI-C6 alkyl, alkoxy, hydroxyalkyl, heteroalkyl, haloalkyl, -L-cycloalkyl, -L-N(R5)2,-Y1-N(R5)2, -L-heteroaryl, -L-heterocyclyl, or -Y1-heterocyclyl. wherein the heteroaryl of the -L-heteroaryl or the heterocyclyl portion of the L-heterocyclyl, or Y1-heterocyclyl is optionally substituted with one or more R7.

[0255] Embodiment 53. The method of Embodiment 50, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is pyrazolyl optionally substituted with one R4independently selected from hydroxyalkyl, heteroalkyl, haloalkyl, -Y^Ci-Ce alkyl, -L-N(R5)2, L-heterocyclyl or L-heteroaryl, wherein the heteroaryl of the L-heteroaryl or the heterocyclyl portion of the L-heterocyclyl is optionally substituted with one or more R7.

[0256] Embodiment 54. The method of Embodiment 53, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-heteroaryl and L is methylene wherein the heteroaryl is pyridyl optional substituted with one or more R7.

[0257] Embodiment 55. The method of Embodiment 53, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-heterocyclyl optionally substituted with one or more R7where L is a bond and the heterocyclyl portion of the L-heterocyclyl is oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, piperazinyl, or 4-methylpiperazinyl.

[0258] Embodiment 56. The method of Embodiment 53, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-heterocyclyl optionally substituted with one or more R7WSGR Ref. 47134-784.601where L is methylene and the heterocyclyl portion of the L-heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl, pyrrolidinone, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, piperazinyl, or 4-methylpiperazinyl.

[0259] Embodiment 57. The method of Embodiment 53, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -L-N(R5)2 where L is methylene and each R5is independently hydrogen, each R5is independently C1-C3 alkyl or one R5is C1-C3 alkyl and one R5is hydrogen.

[0260] Embodiment 58. The method of Embodiment 53, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R4is -Y^Ci-Ce alkyl where Y1is a bond and the Ci-Ce alkyl is methyl, ethyl, or isopropyl.

[0261] Embodiment 59. The method of Embodiment 50, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is pyrazolyl optionally substituted with two R4groups each independently selected from hydroxyalkyl, heteroalkyl, haloalkyl, and -Y^Ci-Ce alkyl.

[0262] Embodiment 60. The method of Embodiment 50, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is pyridyl optionally substituted with one R4independently selected from cyano, halogen, alkoxy, hydroxyalkyl, heteroalkyl, haloalkyl, -Y^Ci-Ce alkyl, -L-N(R5)2,-Y4-N(R5)2, -L-cycloalkyl, or -L-heterocyclyl optionally substituted with one or more R7.

[0263] Embodiment 61. The method of any one of Embodiments 16 to 27, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is -L-cycloalkyl optionally substituted with one or more R4.

[0264] Embodiment 62. The method of any one of Embodiments 16 to 27, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R1is -L-heterocyclyl optionally substituted with one or more R4.

[0265] Embodiment 63. The method of Embodiment 62, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, L is a bond and the heterocyclyl is piperidinyl or tetrahydropyranyl .

[0266] Embodiment 64. The method of any one of Embodiments 1 to 13, wherein the EED inhibitor is a compound selected from the group consisting of:WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601WSGR Ref. 47134-784.601

[0267] Embodiment 65. The method of any one of Embodiments 1 to 13, wherein the EED inhibitor is aWSGR Ref. 47134-784.601and

[0268] Embodiment 66. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.

[0269] Embodiment 67. The method of Embodiment 65, wherein the EED inhibitor is:o'"or a pharmaceutically acceptable salt thereof.

[0270] Embodiment 68. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.WSGR Ref. 47134-784.601

[0271] Embodiment 69. The method of Embodiment 65, wherein the EED inhibitor is:(Compound 1), or a pharmaceutically acceptable salt thereof.

[0272] Embodiment 70. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.

[0273] Embodiment 71. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.

[0274] Embodiment 72. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.WSGR Ref. 47134-784.601

[0275] Embodiment 73. The method of Embodiment 65, wherein the EED inhibitor is:F, or a pharmaceutically acceptable salt thereof.

[0276] Embodiment 74. The method of Embodiment 65, wherein the EED inhibitor is:HNFx^fx ^xLJO , or a pharmaceutically acceptable salt thereof.

[0277] Embodiment 75. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.

[0278] Embodiment 76. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.

[0279] Embodiment 77. The method of Embodiment 65, wherein the EED inhibitor is:or a pharmaceutically acceptable salt thereof.WSGR Ref. 47134-784.601

[0280] Embodiment 78. A method of treating colorectal cancer in a subject, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitorof formula, or a pharmaceutically acceptable salt thereof.

[0281] Embodiment 79. A method of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED)inhibitor of formula, or a pharmaceutically acceptable salt thereof.

[0282] Embodiment 80. A method of treating colorectal cancer in a subject, wherein the colorectal cancer in the subject has been determined to comprise a KRAS G12C mutation by an FDA approved test, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectodermdevelopment (EED) inhibitor of formula, or a pharmaceutically acceptable salt thereof.WSGR Ref. 47134-784.601

[0283] Embodiment 81. A method of treating KRAS G12C-mutated colorectal cancer, comprising administering to the subject (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development(EED) inhibitor of formula, or a pharmaceutically acceptable salt thereof.

[0284] Embodiment 82. The method of any one of Embodiments 78 to 81, wherein the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC-9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400, RMC-4998, D35-001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674, LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, andYL-17231 (TEB-17231).

[0285] Embodiment 83. The method of Embodiment 82, wherein the inhibitor of KRAS G12C is adagrasib or sotorasib.

[0286] Embodiment 84. The method of any one of Embodiments 1 to 83, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject sequentially or simultaneously.

[0287] Embodiment 85. The method of Embodiment 84, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject sequentially.

[0288] Embodiment 86. The method of Embodiment 84, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject simultaneously.

[0289] Embodiment 87. The method of any one of Embodiments 1 to 83, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject on the same day.

[0290] Embodiment 88. The method of any one of Embodiments 1 to 87, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject within a 24-hour period.

[0291] Embodiment 89. The method of any one of Embodiments 1 to 88, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject once per day or twice per day.

[0292] Embodiment 90. The method of Embodiment 89, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject once per day.

[0293] Embodiment 91. The method of Embodiment 89, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject twice per day.WSGR Ref. 47134-784.601

[0294] Embodiment 92. The method of any one of Embodiments 1 to 89, wherein the inhibitor of KRAS G12C is administered to the subject twice per day and the EED inhibitor is administered to the subject once per day.

[0295] Embodiment 93. The method of any one of Embodiments 1 to 89, wherein the inhibitor of KRAS G12C is administered to the subject once per day and the EED inhibitor is administered to the subject twice per day.

[0296] Embodiment 94. The method of any one of Embodiments 1 to 93, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject with food or without food.

[0297] Embodiment 95. The method of Embodiment 94, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject with food.

[0298] Embodiment 96. The method of Embodiment 94, wherein the inhibitor of KRAS G12C and the EED inhibitor are administered to the subject without food.

[0299] Embodiment 97. The method of any one of Embodiments 1 to 96, wherein the colorectal cancer in the subject is selected from metastatic colorectal cancer, locally advanced colorectal cancer, locally advanced metastatic colorectal cancer, non-metastatic colorectal cancer, colon cancer, metastatic colon cancer, locally advanced colon cancer, locally advanced metastatic colon cancer, non-metastatic colon cancer, rectal cancer, metastatic rectal cancer, locally advanced rectal cancer, locally advanced metastatic rectal cancer, non-metastatic rectal cancer.

[0300] Embodiment 98. The method of Embodiment 97, wherein the colorectal cancer in the subject is metastatic colorectal cancer.

[0301] Embodiment 99. The method of Embodiment 97, wherein the colorectal cancer in the subject is locally advanced colorectal cancer.

[0302] Embodiment 100. The method of Embodiment 97, wherein the colorectal cancer in the subject is locally advanced metastatic colorectal cancer.

[0303] Embodiment 101. The method of Embodiment 97, wherein the colorectal cancer in the subject is non-metastatic colorectal cancer.EXAMPLESExample 1: Study of Compound 1 and Adagrasib in a KRAS G12C Human Colorectal Adenocarcinoma Subcutaneous Xenograft Model

[0304] Female NOD SCID mice (6 to 8 weeks old; GemPharmatech Biotech CO, Ltd.) were inoculated subcutaneously on the right flank with 5xl06SW1463 tumor cells in 0.1 mL of medium and Matrigel mixture (1:1 ratio) for tumor development. After the mean tumor volume in the animals reached 154 mm3, they were randomly assigned to 4 treatment groups: (a) vehicle; (b) Compound 1 ; (c) adagrasib; and (d) Compound 1 plus adagrasib. The treatments were administered to the tumor-bearing mice according to the study design set forth in Table 1. For the combination treatments, adagrasib was dosed first and 2 hours later Compound 1 was administered.Table 1WSGR Ref. 47134-784.601Treatment No. of Dosing vol. Dosing frequency Group Dose level (mg / kg)Group mice (ul / g) and duration 1 Vehicle 10 NA 5 QD (22 days) 2 Compound 1 10 100 5 QD (19 days) 3 Adagrasib 10 30 5 QD (44 days) Compound 1 Compound 1 (100);4 10 5 QD (44 days) plus adagrasib adagrasib (30)

[0305] Subcutaneous tumor volumes were measured twice a week by caliper using the formula: Tumor Volume (TV) = (length x width2) / 2. Mouse body weights were measured daily with a weighing scale. Conditions of animal health and clinical signs of side effects were monitored by daily observation of gross morphology and necropsy of euthanized animals at study endpoint. The study was terminated following 21 to 28 days of treatment as defined in the study protocol. Tumor growth inhibition (TGI) was calculated as [1 - (TVtf - TVtO) / (TVcf -TVcO)] x 100%, where TVtf was the tumor volume (TV) mean of treatment group at final or last treatment day, TVtO was the TV mean of treatment group at treatment day 0, TVcf was the TV mean of the control group at final or last treatment day and TVcO was the TV mean of the control group at treatment day 0. As shown in FIG. 1 : (a) the vehicle and Compound 1 treatment groups exhibited no detectable TGI; (b) the adagrasib treatment group exhibited 96% TGI as determined on treatment day 21 with no tumor regressions; and (c) the Compound 1 plus adagrasib treatment group exhibited 101% TGI as determined on treatment day 21 , with 4 of 10 mice exhibiting tumor regression. As also shown in FIG. 1 , the combination of Compound 1 plus adagrasib exhibited prolonged tumor growth inhibition compared to the Compound 1 monotherapy and adagrasib monotherapy groups. Individual tumor volumes at day 18 for individual animal (and mean± SEM) from each treatment group are shown in FIG. 2. Individual tumor volumes at day 42 for each animal (and mean ± SEM) in the adagrasib monotherapy and Compound 1 plus adagrasib treatment groups are shown in FIG. 3. As shown in FIG. 2 and FIG.3 , the Compound 1 plus adagrasib treatment group exhibited the best overall TGI amongst all the treatment groups.

Claims

WSGR Ref. 47134-784.601CLAIMS WHAT IS CLAIMED is:

1. A method of treating colorectal cancer in a subj ect, comprising administering to the subj ect (a) an inhibitor of KRAS G12C, and (b) an embryonic ectoderm development (EED) inhibitor, wherein the colorectal cancer has been determined to comprise a KRAS G12C mutation.

2. The method of claim 1, wherein the subject has not acquired one or more secondary site pathway mutations.

3. The method of claim 2, wherein the secondary site pathway mutation isa RAS / MAPK pathway alteration, optionally a KRAS alternation,a KRAS-activating mutation, optionally a mutation in codon 12 or 13,a KRAS amplification,a RAF / MAPK pathway mutation or fusion, optionally MAP2K1K57and BRAFV600E, a KRAS switch-II pocket mutation, optionally KRAS Y96D,an NRAS / HRAS mutation, optionally a single nucleotide variant in codon 61, a RTK amplification, capable of re-activating or mediating reactivation of a RAS / MAPK signaling pathway, optionally wherein the RTK amplification is MET amplification, ERBB2 amplification, ERBB4 amplification, EGFR amplification, or FGFR2 amplification, ora RTK fusion, capable of re-activating or mediating reactivation of a RAS / MAPK signaling pathway, optionally wherein the RTK fusion is CCDC6-RET fusion, EML4-ALK fusion, FGFR3-TACC fusion, AHCYL2-MET fusion, MET-SLC12A9 fusion, or GOPC-ROS1 fusion, orany combination thereof.

4. The method of claim 3, wherein:the KRAS-activating mutation is KRAS G12A, KRAS G12D, KRAS G12V, KRAS G13D, or KRAS Q61H;the KRAS switch-II pocket mutation is KRAS Y64 mutation, KRAS H95 mutation, KRAS Y96 mutation, optionally R68S, M72T, H95R, H95N, H95L, H95Q, Y96D, Y96N, Y96H, or Q99L; orthe NRAS mutation is NRAS Q61K, or NRAS Q61R.

5. The method of any one of claims 1 to 4, wherein the inhibitor of KRAS G12C is selected from adagrasib, sotorasib, divarasib (GDC-6036), olomorasib (LY3537982), garsorasib (D-1553), glecirasib (JAB-21822), MK-1084, JNJ-74699157 (ARS-3248), GFH925, ZG19018, YL-15293, HBI-2438, FMC-376, HS-10370, BBO-8520, elironrasib (RMC-6291), KQB365, ARS-1620, 1_AM, ARS-853, BI-1823911, JDQ443, BPI-421286, GH35, BEBT-607, JAB-21000, LY3499446, SY-5933, HRS-7058, GEC255, daraxonrasib (RMC-6236), zoldonrasib (RMC- 9805), RMC-7977, ADT-007, MRTX-1133, BI-2852, JAB-23400 RMC-4998, D35- 001, fulzerasib (GFH925, IBI351), opnurasib (JDQ-443), HYP-2090PTSA, BI-3706674,WSGR Ref. 47134-784.601LY4066434, ALTA-3263, PF-07934040, BGB-53038, PF-07985045, QTX3034, QTX3544, LUNA18, RM-018, andYL-17231 (TEB-17231).

6. The method of claim 5, wherein the inhibitor of KRAS G12C is adagrasib or sotorasib.

7. The method of any one of claims 1 to 6, wherein the EED inhibitor is a small molecule having a molecular weight of less or equal to 3000 Daltons.

8. The method of any one of claims 1 to 7, wherein the EED inhibitor is selected from EED226, A- 395, APG-5918, BR-001, BR-002, EEDi-5285, EEDi-1056, FTX-6274, pociredir (FTX-6058), HJM-353, and MAK683.

9. The method of any one of claims 1 to 7, wherein the EED inhibitor is a compound of Formula (I) R1Formula (I)or a pharmaceutically acceptable salt thereof:wherein:- represents a single or a double bond;Z is O or S;X is O, CR5, CR5OH, or C(R5)2, wherein:when X is O, - is a single bond;when X is C(R5)2, - is a single bond;when X is CR5OH, is a single bond; orwhen X is CR5, is a double bond;R1is aryl, heteroaryl, L-cycloalkyl, -N(R5)heterocyclyl, or L-heterocyclyl, wherein the aryl, the heteroaryl or the cyclyl portion of the L-cycloalkyl, -N(R5)heterocyclyl, or L-heterocyclyl is optionally substituted with one or more R4;R2is cyano, -COOR5, -C(O)N(R5)2, or -C(O)N(R5)2 wherein each R5taken together with the nitrogen atom to which they are attached form a 5 - 8 membered heterocyclic ring optionally substituted with one or more R4;each R3is independently C1-C3 alkyl or halogen;each R4is independently oxo, cyano, halogen, -PCLCCi-Cs alkyl)2, hydroxyl, alkoxy, hydroxyalkyl, heteroalkyl, aralkyl, haloalkyl, -COOR5, -Y2-haloalkyl, -Y^Ci-Ce alkyl, -Y2-Ci-Ce alkyl, -L-cycloalkyl, -L-heteroaryl, -L-heterocyclyl, -Y'-heterocyclyl. -Y2-heterocyclyl, -L- N(R5)2, -O-L-N(R5)2, -C(CF3)N(R5)2, -Y1-N(R5)2, or -Y2-N(R5)2, wherein the ring portion of theWSGR Ref. 47134-784.601aralkyl, -L-cycloalkyl, -L-heteroaryl, -L-heterocyclyl or -Y1-heterocycly I is optionally substituted with one or more R7;L is a bond or C1-C4 alkylene;Y1is a bond, -C(O)-, or -NHC(O)-;Y2is a bond, -S-, -SO-, -SO2-, or -NR5SO2-,each R5is hydrogen or C1-C3 alkyl;R6is hydrogen, C1-C3 alkyl, halogen, haloalkyl, hydroxyalkyl, or heteroalkyl;each R7is oxo, cyano, hydroxyl, alkoxy, halogen, haloalkyl, hydroxyalkyl, heteroalkyl, cycloalkyl, -L-N(R5)2, Ci-Ce alkyl, or -Y1-heterocycly 1; andn is 1 or 2.

10. The method of claim 9, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, Z is O.

11. The method of claim 9 or claim 10, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, n is 1.

12. The method of any one of claims 9 to 11, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, R2is cyano.

13. The method of any one of claims 9 to 12, wherein in the compound of Formula (I), or a pharmaceutically acceptable salt thereof, X is C(R5)2 and - is a single bond.

14. The method of any one of claims 1 to 13, wherein the EED inhibitor is a compound selected fromWSGR Ref. 47134-784.60115.pharmaceutically acceptable salt thereof.

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