Improved synthesis of kras g12c inhibitor compound

Abstract

This disclosure relates to an improved, efficient, and scalable method for preparing intermediate compounds, such as 2,2',2''-(1,3,5,2,4,6-trioxaborane-2,4,6-triyl)tri(3-fluorophenol), which can be used to synthesize compounds for treating cancers with KRAS G12C mutations, such as compound 9.

Description

Technical Field

[0001] The present disclosure relates to an improved, efficient, and scalable method for preparing a structured The intermediate compound, such as the compound having Formula 6A, can be used to synthesize a compound for treating cancers with KRAS G12C mutation. Prior Art

[0002] KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and bile duct cancer. KRAS mutations are also observed in approximately 25% of NSCLC patients, and some studies have shown that KRAS mutations are negative prognostic factors in NSCLC patients. Recently, V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations have been found to confer resistance to epidermal growth factor receptor (EGFR)-targeted therapies in colorectal cancer; therefore, KRAS mutation status can provide important information before prescribing TKI therapy. Overall, new medical treatments are needed for patients with pancreatic, lung adenocarcinoma, or colorectal cancer, particularly those diagnosed with such cancers characterized by KRAS mutations, including those who have progressed after chemotherapy. Summary of the Invention

[0003] The present disclosure relates to improved preparations of compounds having the following chemical structure: . Simple diagram description

[0004] none Implementation Method

[0005] [Cross-reference to related applications] [] This application claims the benefit of U.S. Provisional Application No. 62 / 935,502, filed November 14, 2019, which is incorporated herein by reference in its entirety. [definition] [] Abbreviations: The following abbreviations may be used in this document: ACN Acetonitrile AcOH Acetic acid aq or aq. Water-based BOC or Boc tertiary butoxycarbonyl BuOH n-Butanol BuOAc Butyl acetate cpme Cyclopentyl methyl ether CHCl 3 Chloroform DCE 1,2-Dichloroethane DABCO 1,4-diazabicyclo[2.2.2]octane DCM dichloromethane DMA N,N-dimethylacetamide DMAP 4-Dimethylaminopyridine DME 1,2-Dimethoxyethane DMF N,N-dimethylformamide DMSO Dimethyl sulfoxide Dppf, DPPF or dppf 1,1'-Bis(diphenylphosphino)ferrocene eq or eq. or equiv. equivalent ESI or ES Electrospray ionization Et Ethyl Et2O Ether EtOAc Ethyl acetate EtOH ethanol g gram h Hour H 20 water HPLC High-pressure liquid chromatography iPr Isopropyl IPA Isopropyl alcohol IPAc Isopropyl acetate iPr 2NEt or DIPEA N-Ethyldiisopropylamine (Hünig's base) KHMDS Potassium hexamethyldisilazide KOAc Potassium acetate LDA Lithium diisopropylamide Lawesson's reagent 2,4-Bis(4-methoxyphenyl)-2,4-dithio-1,3,2,4-dithiadiphosphatane, 2,4-bis(4-methoxyphenyl)-1,3-dithia-2,4-diphosphatane 2,4-disulfide LC MS, LCMS, LC-MS, or LC / MS Liquid chromatography-mass spectrometry LG Leaving groups (e.g., halogen, mesylate, triflate) LHMDS or LiHMDS Lithium hexamethyldisilazide m / z mass-to-charge ratio Me methyl MeCN Acetonitrile MeOH Methanol Met Metal species for cross-coupling (e.g., MgX, ZnX, SnR3, SiR3, B(OR)2) 2-MeTHF 2-Methyltetrahydrofuran mg mg min minute MIBK 4-Methyl-2-pentanone mL milliliters MS Mass spectrometry MTBE Methyl tertiary butyl ether n-BuLi n-Butyllithium NaHMDS Sodium hexamethyldisilazide NBS N-Bromosuccinimide NCS N-Chlorosuccinimide NMR Nuclear magnetic resonance imaging Pd 2(dba) 3 Tris(dibenzylideneacetone)dipalladium(0) Pd(dppf)Cl 2·DCM [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane Pd(PPh 3) 4 Tetrakis(triphenylphosphine)palladium(0) Ph Phenyl PR or PG or Prot. group Protecting groups rbf round-bottom flask RP-HPLC Reversed-phase high-performance liquid chromatography RT or rt Room temperature sat. or satd. Saturated SFC Supercritical liquid chromatography SPhos Pd G3 or SPhos G3 (2-Dicyclohexylphosphino-2′,6′-dimethoxybiphenyl)[2-(2′-amino-1,1′-biphenyl)]methanesulfonate palladium(II) TBAF Tetra-n-butylammonium fluoride TBTU N,N,N',N'-Tetramethyl-O-(benzotriazol-1-yl)uronium tetrafluoroborate t-BuOH Tertiary Butanol TEA or Et 3N Trimethylamine TFA Trifluoroacetic acid THF Tetrahydrofuran UV ultraviolet light XRPD X-ray powder diffraction

[0006] Unless otherwise indicated, the use of the terms "a," "an," "the," and similar referents in the context of describing the invention (especially in the context of the claims) are to be construed to cover both the singular and the plural, unless otherwise indicated. Recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, and each separate value is incorporated into the specification as if it were individually recited herein. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise indicated. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0007] As used herein, the term "alkyl" refers to straight-chain and branched C1-C8 hydrocarbon groups, including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, secondary butyl, tertiary butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, and 2-ethylbutyl. The term Cmn refers to an alkyl group having from "m" to "n" carbon atoms. The term "alkylene" refers to an alkyl group having a substituent. Alkyl (e.g., methyl) or alkylene (e.g., -CH2-) groups may be substituted with one or more, and typically one to three, groups independently selected from, for example, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, -NC, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C10 cycloalkyl, C3-C10 heterocycloalkyl, C5-C10 aryl, and C5-C10 heteroaryl. The term "haloalkyl" specifically refers to an alkyl group in which at least one (e.g., 1 to 6) or all hydrogen atoms of the alkyl group are replaced by halo atoms.

[0008] The terms "alkenyl" and "alkynyl" refer to an alkyl group further including a double bond or a triple bond, respectively.

[0009] As used herein, the term "halo" refers to fluoro, chloro, bromo, and iodo. The term "alkoxy" is defined as -OR, where R is alkyl.

[0010] As used herein, the terms "amino" or "amine" interchangeably refer to an -NR2 group, where each R is, for example, H or a substituent. In some embodiments, the amino group is further substituted to form an ammonium ion, such as NR3+. Ammonium moieties are specifically included in the definition of "amino" or "amine." Substituents can be, for example, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, amide, or formate. The R group can be further substituted, for example, with one or more (e.g., one to four) groups selected from the group consisting of halo, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, formate, amine, and amide. An "amide" or "amido" group interchangeably refers to a group similar to an amine or amino group but additionally includes C(O), such as -C(O)NR2.

[0011] As used herein, the term "aryl" refers to a C6-14 monocyclic or polycyclic aromatic group, preferably a C6-10 monocyclic or bicyclic aromatic group, or a C10-14 polycyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthracenyl, phenanthrenyl, pyrenyl, biphenyl, and terphenyl. Aryl also refers to C10-14 bicyclic and tricyclic carbocycles in which one ring system is aromatic and the other ring systems are saturated, partially unsaturated, or aromatic, such as dihydronaphthyl, indenyl, dihydroindenyl, or tetrahydronaphthyl (tetrahydronaphthyl). Unless otherwise indicated, an aryl group may be unsubstituted or substituted by one or more, and in particular one to four, groups independently selected from, for example, halo, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, -CF 3 , -OCF 3 , -NO 2 , -CN, -NC, -OH, alkoxy, amino, -CO 2H, -CO 2C 1-C 8 alkyl, -OCOC 1-C 8 alkyl, C 3-C 10 cycloalkyl, C 3-C 10 heterocycloalkyl, C 5-C 10 aryl and C 5-C 10 heteroaryl.

[0012] As used herein, the term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic carbocyclic ring, wherein the polycyclic rings may be fused, bridged, or spirocyclic. The carbocyclic ring may have 3 to 10 carbon ring atoms. Contemplated carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclononyl.

[0013] As used herein, the term "heterocycloalkyl" refers to a monocyclic or polycyclic (e.g., bicyclic), saturated or partially unsaturated ring system containing 3 or more (e.g., 3 to 12, 4 to 10, 4 to 8, or 5 to 7) total atoms, of which 1-5 (e.g., 1, 2, 3, 4, or 5) atoms are independently selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heterocycloalkyl groups include azetidinyl, pyrrolidinyl, piperidinyl, piperonyl, dihydropyrrolyl, oxepinyl, dioxepinyl, thiepanyl, and diazepanyl.

[0014] Unless otherwise indicated, a cycloalkyl or heterocycloalkyl group may be unsubstituted or substituted with one or more, and particularly one to four, groups. Some contemplated substituents include halo, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, -OCF 3, -NO 2, -CN, -NC, -OH, alkoxy, amine, -CO 2H, -CO 2C 1-C 8 alkyl, -OCOC 1-C 8 alkyl, C 3-C 10 cycloalkyl, C 3-C 10 heterocycloalkyl, C 5-C 10 aryl, and C 5-C 10 heteroaryl.

[0015] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic ring system (e.g., bicyclic) containing one to three aromatic rings and one to four (e.g., 1, 2, 3, or 4) heteroatoms selected from nitrogen, oxygen, and sulfur in the aromatic rings. In certain embodiments, heteroaryl groups have 5 to 20, 5 to 15, 5 to 10, or 5 to 7 ring atoms. Heteroaryl also refers to C10-14 bicyclic and tricyclic rings in which one ring system is aromatic and the other ring systems are saturated, partially unsaturated, or aromatic. Examples of heteroaryl groups include, but are not limited to, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrimidinyl, pyrazolyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazolyl, triazolyl, benzofuranyl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzothio The heteroaryl radicals include, but are not limited to, benzotriazolyl, benzoxazolyl, furopyridinyl, imidazopyridinyl, imidazothiazolyl, indolyl, indolyl, indazolyl, isobenzofuranyl, isobenzothiophenyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalinyl, pteridinyl, purinyl, pyridopyridinyl, pyrrolopyridinyl, quinolinyl, quinazolinyl, thiadiazolopyrimidinyl, and thienopyridinyl. Unless otherwise indicated, heteroaryl radicals may be unsubstituted or substituted with one or more, and in particular one to four, or one or two, substituents. Contemplated substituents include halo, C 1- 8 alkyl, C 2- 8 alkenyl, C 2- 8 alkynyl, -OCF 3, -NO 2, -CN, -NC, -OH, alkoxy, amine, -CO 2H, -CO 2C 1-C 8 alkyl, -OCOC 1-C 8 alkyl, C 3-C 10 cycloalkyl, C 3-C 10 heterocycloalkyl, C 5-C 10 aryl, and C 5-C 10 heteroaryl.

[0016] As used herein, the term Boc refers to the structure . [] [Specific implementation method] [] [Implementation Method] [1] []

[0017] In one embodiment of the present disclosure, the present disclosure comprises a compound having Formula 6A: . [] [Implementation Method] [2] []

[0018] In another embodiment of the present disclosure, the present disclosure comprises a composition comprising a compound having Formula 6A: . [] [Implementation Method] [3] []

[0019] In another embodiment of the present disclosure, the present disclosure includes a method for preparing a compound having Formula 6A: , The method comprises making a compound having the structure: The mixture is reacted with an acid and at least one solvent. [] [Implementation Method] [4] []

[0020] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 3, wherein the acid is BBr 3. [] [Implementation Method] [5] []

[0021] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 3, wherein the at least one solvent is dichloromethane. [] [Implementation Method] [6] []

[0022] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 3, wherein the at least one solvent is heptane. [] [Implementation Method] [7] []

[0023] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 3, wherein the mixture is cooled to about -20°C. [] [Implementation Method] [8] []

[0024] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 3, wherein the method is to prepare a compound having the following structure: , including compounds having the following structure: Mix with reagents, primary base, secondary amine base, catalyst, and acid. [] [Implementation Method] [9] []

[0025] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 8, wherein the first base is n-butyllithium. [] [Implementation Method]

[10] []

[0026] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 8, wherein the secondary amine base is diisopropylamine. [] [Implementation Method]

[11] []

[0027] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 8, wherein the catalyst is triethylamine hydrochloride. [] [Implementation Method]

[12] []

[0028] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 8, wherein the reagent is triethyl borate. [] [Implementation Method]

[13] []

[0029] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 8, wherein the acid is HCl. [] [Implementation Method]

[14] []

[0030] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 3, wherein the compound of formula 6A is used to produce a compound of formula 7: . [] [Implementation Method]

[15] []

[0031] In another embodiment of the present disclosure, the present disclosure includes a method for preparing a compound having Formula 7: , comprising making a compound having Formula 6A: , With a compound having formula 6: , The steps of the reaction in the presence of Pd(dpePhos)Cl2 and KOAc. [] [Implementation Method]

[16] []

[0032] In another embodiment of the present disclosure, the present disclosure includes the method as described in Embodiment 3, wherein the compound of Formula 6A is used to produce a compound of Formula 9: 9. [] [Implementation Method]

[17] []

[0033] In another embodiment of the present disclosure, the present disclosure includes the method as described in embodiment 16, wherein the method further comprises mixing the compound of formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition. [] [Compounds of the Present Disclosure] []

[0034] Provided herein are KRAS inhibitors having structures discussed in more detail below.

[0035] The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds, wherein one or more atoms of the disclosed compounds are 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 that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as 2H, 3H, 11C, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 31P, 32P, 35S, 18F, 36Cl, 123I, and 125I, respectively. These radiolabeled compounds can be used to determine or measure the effectiveness of the compounds by characterizing, for example, the site or mode of action, or binding affinity to a pharmacologically important site of action. Certain isotopically-labeled compounds of the present disclosure (e.g., those incorporating a radioactive isotope) are useful in drug and / or substrate tissue distribution studies. The radioactive isotopes tritium (i.e., 3H) and carbon-14 (i.e., 14C) are particularly useful for this purpose in view of their ease of incorporation and ready-to-use detection formats.

[0036] Substitution with heavier isotopes (such as deuterium, ie, 2 H) may afford certain therapeutic advantages resulting from greater metabolic stability (eg, increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances.

[0037] Substitution with positron-emitting isotopes (e.g., 11C, 18F, 15O, and 13N) can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art, or by methods analogous to those described in the Preparations and Examples described below, using an appropriate isotopically labeled reagent in place of the unlabeled reagent previously employed.

[0038] Isotopically labeled compounds disclosed herein can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Schemes, using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously employed.

[0039] Certain compounds disclosed herein may exist as stereoisomers (i.e., isomers that differ only in the arrangement of their atoms in space), including optical isomers and conformers (or conformers). The compounds disclosed herein include all stereoisomers as pure preparations of individual stereoisomers and enriched preparations of each, as well as racemic mixtures of such stereoisomers, and individual diastereomers and enantiomers that can be separated according to methods known to those skilled in the art. In addition, the compounds disclosed herein include all tautomeric forms of such compounds.

[0040] Certain compounds disclosed herein may exist as atropisomers, which are conformational stereoisomers that occur when rotation about a single bond in a molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers as pure preparations of individual atropisomers, enriched preparations of each, or non-specific mixtures of each. If the barrier to rotation about a single bond is high enough and the interconversion between conformations is slow enough, then separation and isolation of the isomeric species may be permitted. For example, groups such as, but not limited to, the following groups 、 、 and May exhibit restricted rotation.

[0041] The term "monohydrate" refers to a salt of Compound 9 that has approximately one associated water molecule. Those skilled in the art will appreciate that the exact number of associated water molecules can vary slightly at any given time due to variable temperature, pressure, and other environmental influences. All minor variations in the number of associated water molecules are contemplated within the scope of the present disclosure.

[0042] The term "dihydrate" refers to a salt of Compound 9 that has approximately two associated water molecules. Those skilled in the art will appreciate that the exact number of associated water molecules can vary slightly at any given time due to variable temperature, pressure, and other environmental influences. All minor variations in the number of associated water molecules are contemplated within the scope of the present invention.

[0043] The term "co-crystal" refers to a crystalline material comprising two or more compounds at ambient temperature (20°C to 25°C, preferably 20°C), at least two of which are held together by weak interactions, wherein at least one of the compounds is a co-crystal former and another is Compound 5. Weak interactions are defined as interactions that are neither ionic nor covalent, and include, for example, hydrogen bonds, van der Waals forces, and π-π interactions.

[0044] The term "amorphous form" or "amorphous" refers to a material that lacks long-range order and therefore does not exhibit distinct X-ray diffraction peaks (ie, Bragg diffraction peaks). An XRPD pattern of an amorphous material is characterized by one or more amorphous halos.

[0045] The term "amorphous halo" refers to the approximately bell-shaped maximum in the X-ray powder pattern of an amorphous material.

[0046] The term "substantially pure" refers to a solid form of Compound 9 having a purity greater than about 95%, specifically greater than about 99.5%, more specifically greater than about 99.8%, and still more specifically greater than about 99.9%.

[0047] The term "patient" refers to animals such as dogs, cats, cows, horses, sheep, and humans. Certain patients are mammals. The term "patient" includes both male and female patients.

[0048] The terms "treating," "treat," or "treatment" and such include preventative (eg, prophylactic) and palliative treatment.

[0049] The term "excipient" means any pharmaceutically acceptable additive, carrier, diluent, adjuvant, or other ingredient, other than the active pharmaceutical ingredient (API), that is typically incorporated for formulation and / or administration to a patient. [] [Drug composition, administration and route of administration] []

[0050] Also provided herein are pharmaceutical compositions comprising a compound as disclosed herein and a pharmaceutically acceptable excipient, such as a diluent or carrier. Compounds and pharmaceutical compositions suitable for use in the present invention include those that can be administered in an effective amount to achieve their intended purpose. Administration of the compounds is described in more detail below.

[0051] Suitable pharmaceutical formulations can be determined by a skilled artisan based on the route of administration and desired dosage. See, for example, Remington's Pharmaceutical Sciences, 1435-712 (18th ed., Mack Publishing Co., Easton, Pennsylvania, 1990). The formulation can affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the administered agent. Depending on the route of administration, a suitable dosage can be calculated based on body weight, body surface area, or organ size. Further refinement of the calculations required to determine an appropriate therapeutic dosage can be routinely performed by those skilled in the art without undue experimentation, particularly based on the dosage information and assays disclosed herein and pharmacokinetic data available from animal or human clinical trials.

[0052] The phrases "pharmaceutically acceptable" or "pharmacologically acceptable" refer to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to animals or humans. As used herein, "pharmaceutically acceptable" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such excipients with pharmaceutically active substances is well known in the art. Unless any conventional media or agents are incompatible with the therapeutic composition, their use in the therapeutic composition is contemplated. Supplementary active ingredients may also be incorporated into the composition. In an exemplary embodiment, the formulation may include corn syrup solids, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tricalcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dipotassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, Calcium carbonate, L-glutamate, L-cystine dihydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, thiamin sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0053] The compound may be present in the pharmaceutical composition as a pharmaceutically acceptable salt. As used herein, "pharmaceutically acceptable salt" includes, for example, base addition salts and acid addition salts.

[0054] Pharmaceutically acceptable base addition salts can be formed with metals or amines, such as alkali metals and alkaline earth metals or organic amines. Pharmaceutically acceptable salts of the compounds can also be prepared with pharmaceutically acceptable cations. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkali metal cations, alkaline earth metal cations, ammonium cations, and quaternary ammonium cations. Carbonates or bicarbonates are also possible. Examples of metals used as cations include sodium, potassium, magnesium, ammonium, calcium, or ferric iron. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine.

[0055] Pharmaceutically acceptable acid addition salts include inorganic or organic acid salts. Examples of suitable acid salts include hydrochlorides, formates, acetates, citrates, salicylates, nitrates, and phosphates. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include, for example, formic acid, acetic acid, citric acid, oxalic acid, tartaric acid or mandelic acid, hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic formic, sulfonic, sulfonic or phosphoric acid or N-substituted aminosulfonic acids, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucuronic acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2- Salts of phenoxybenzoic acid, 2-acetyloxybenzoic acid, pamoic acid, nicotinic acid or isonicotinic acid; and salts with amino acids, such as the 20 alpha amino acids involved in protein synthesis in nature, such as glutamate or aspartic acid, as well as with phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane 1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene 2-sulfonic acid, naphthalene 1,5-disulfonic acid, 2-phosphoglyceric acid or 3-phosphoglyceric acid, glucose 6-phosphate, N-cyclohexylaminosulfonic acid (for the formation of cyclohexylaminosulfonic acid salts), or with other acidic organic compounds, such as ascorbic acid.

[0056] Pharmaceutical compositions containing the compounds disclosed herein can be manufactured in a conventional manner, for example, by conventional mixing, dissolving, granulating, dragee making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation depends on the chosen route of administration.

[0057] For oral administration, suitable compositions can be readily formulated by combining the compounds disclosed herein with pharmaceutically acceptable excipients (e.g., carriers) well known in the art. Such excipients and carriers enable the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like for oral ingestion by the patient to be treated. Pharmaceutical formulations for oral use can be obtained by adding solid excipients to the compounds disclosed herein, grinding the resulting mixture as necessary, and processing the granular mixture after adding suitable adjuvants (if desired) to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. If desired, a disintegrant may be added. Pharmaceutically acceptable ingredients for various types of formulations are well known and may include, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coating materials, preservatives, dyes, thickeners, adjuvants, antimicrobial agents, antioxidants, and carriers for various formulation types.

[0058] When a therapeutically effective amount of a compound disclosed herein is administered orally, the composition is typically in the form of a solid (e.g., tablet, capsule, pill, powder, or lozenge) or a liquid formulation (e.g., aqueous suspension, solution, elixir, or syrup).

[0059] When administered in tablet form, the composition may additionally contain a functional solid and / or solid carrier, such as gelatin or an adjuvant. Tablets, capsules, and powders may contain from about 1% to about 95% of the compound, and preferably from about 15% to about 90% of the compound.

[0060] When administered in liquid or suspension form, a functional liquid and / or liquid carrier, such as water, petroleum, or oils of animal or plant origin, may be added. Liquid forms of the composition may further contain physiological saline solution, sugar alcohol solutions, dextrose or other sugar solutions, or glycols. When administered in liquid or suspension form, the composition may contain from about 0.5% to about 90% by weight of a compound disclosed herein, and preferably from about 1% to about 50% by weight of a compound disclosed herein. In one contemplated embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition may be supplied as a rapidly dissolving solid formulation for dissolution or suspension immediately prior to administration.

[0061] When a therapeutically effective amount of the compounds disclosed herein is administered by intravenous, transdermal, or subcutaneous injection, the composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions should fully consider pH, isotonicity, stability, and other factors and is within the skill of the art. Preferred compositions for intravenous, transdermal, or subcutaneous injection, in addition to the compounds disclosed herein, typically contain an isotonic vehicle. Such compositions can be prepared for administration as solutions of the free base or pharmacologically acceptable salt in water, suitably mixed with a surfactant (e.g., hydroxypropylcellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as in oils. Under ordinary conditions of storage and use, such preparations may optionally contain a preservative to prevent microbial growth.

[0062] Injectable compositions may include sterile aqueous solutions, suspensions, or dispersions, as well as sterile powders for the extemporaneous preparation of sterile injectable solutions, suspensions, or dispersions. In all embodiments, the form must be sterile and fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be protected against the contaminating action of microorganisms (e.g., bacteria and fungi) by optionally including a preservative. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils. In one contemplated embodiment, the carrier is non-aqueous or substantially non-aqueous. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin; by maintaining the desired particle size of the compound in the case of dispersions; and by the use of a surfactant. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many embodiments, it will be advantageous to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the composition of absorption delaying agents, for example, aluminum monostearate and gelatin.

[0063] Sterile injectable solutions are prepared by incorporating the active compound in the required amount into an appropriate solvent optionally containing various other ingredients enumerated above, followed by filtered sterilization. Dispersions are generally prepared by incorporating the various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and the required other ingredients from those enumerated above. In the embodiment of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying techniques, which yield a powder of the active ingredient plus any required other ingredients from a previously sterile-filtered solution thereof.

[0064] Slow-release or sustained-release formulations can also be prepared to achieve controlled release of the active compound in contact with body fluids in the gastrointestinal tract and provide a substantially constant and effective level of the active compound in the plasma. For example, release can be controlled by one or more of dissolution, diffusion, and ion exchange. Furthermore, slow-release methods can promote absorption via saturable or restricted pathways within the gastrointestinal tract. For example, for this purpose, the compound can be embedded in a polymer matrix of a biodegradable polymer, a water-soluble polymer, or a mixture of both, and, if necessary, a suitable surfactant. In this context, embedding can mean incorporating microparticles into the polymer matrix. Controlled-release formulations can also be achieved by encapsulating dispersed microparticles or emulsified droplets using known dispersion or emulsion coating techniques.

[0065] For administration by inhalation, the compounds of the present disclosure are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant. 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, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0066] The compounds disclosed herein can be formulated for parenteral administration by injection (e.g., by bolus injection or continuous infusion). Injectable formulations can be presented in unit dosage form (e.g., in ampoules or in multi-dose containers) with added preservatives. The compositions can take the form of, for example, suspensions, solutions, or emulsions in oily or aqueous vehicles and may contain formulatory agents such as suspending agents, stabilizers, and / or dispersing agents.

[0067] Pharmaceutical formulations for parenteral administration include aqueous solutions of the compound in water-soluble form. Alternatively, suspensions of the compound can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound and allow for the preparation of highly concentrated solutions. Alternatively, the compositions of the present invention may be in powder form for constitution with a suitable vehicle (e.g., sterile, pyrogen-free water) prior to use.

[0068] The compounds disclosed herein can also be formulated in rectal compositions, such as suppositories or retention enemas (e.g., containing conventional suppository bases). In addition to the formulations described previously, the compounds can also be formulated as long-acting preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds can be formulated with a suitable polymeric or hydrophobic material (e.g., as an emulsion in an acceptable oil) or ion exchange resin, or as a sparingly soluble derivative (e.g., as a sparingly soluble salt).

[0069] In particular, the compounds disclosed herein can be administered orally, buccally, or sublingually in the form of tablets containing excipients (e.g., starch or lactose), or in capsules or ovules, alone or mixed with excipients, or in the form of elixirs or suspensions containing flavorings or coloring agents. Such liquid preparations can be prepared with pharmaceutically acceptable additives (e.g., suspending agents). The compounds can also be injected parenterally, for example, intravenously, intramuscularly, subcutaneously, or intracoronarily. For parenteral administration, the compounds are best administered in the form of sterile aqueous solutions, which may contain other substances, such as salts or sugar alcohols (e.g., mannitol) or glucose, to render the solution isotonic with the blood.

[0070] For veterinary use, the compounds disclosed herein are administered in a suitably acceptable formulation according to normal veterinary practice. A veterinarian can readily determine the most suitable dosing regimen and route of administration for a particular animal.

[0071] In some embodiments, all necessary components for treating a KRAS-related disorder using a compound as disclosed herein (alone or in combination with another agent or intervention traditionally used to treat such a disease) can be packaged into a kit. Specifically, the present disclosure provides kits for therapeutic interventions for a disease, comprising a packaged set of pharmaceuticals comprising a compound as disclosed herein, along with a buffer and other components for preparing a deliverable form of the pharmaceutical; and / or a device for delivering such pharmaceuticals; and / or any pharmaceutical agents for combination therapy with the compounds disclosed herein; and / or instructions for treating the disease packaged with the pharmaceuticals. The instructions can be fixed in any tangible medium, such as printed paper, or computer-readable magnetic or optical media, or instructions referencing a remote computer data source, such as a World Wide Web page accessible via the Internet.

[0072] A "therapeutically effective amount" means an amount effective to treat or prevent the development of, or alleviate, existing symptoms in the subject being treated. Determination of an effective amount is well within the capabilities of those skilled in the art, particularly in light of the detailed disclosure provided herein. Generally, a "therapeutically effective dose" refers to an amount of a compound that results in a desired effect. For example, in a preferred embodiment, a therapeutically effective amount of a compound disclosed herein reduces KRAS activity by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%, compared to a control.

[0073] The amount of compound administered may depend on the subject being treated, the subject's age, health, sex, and weight, the type of concurrent treatment (if any), the severity of the condition, the nature of the desired effect, the manner and frequency of treatment, and the judgment of the prescribing physician. The frequency of dosing may also depend on the pharmacodynamic effect on arterial oxygen tension. However, the optimal dosage can be adjusted for individual subjects, as will be understood by those skilled in the art and can be determined without undue experimentation. This will generally involve adjusting the standard dosage (e.g., reducing the dosage if the patient is underweight).

[0074] While individual needs vary, determining the optimal range of effective amounts of a compound is within the skill of the art. For administration to humans in the curative or prophylactic treatment of the conditions and disorders identified herein, for example, a typical dosage of a compound of the disclosure can be about 0.05 mg / kg / day to about 50 mg / kg / day, such as at least 0.05 mg / kg, at least 0.08 mg / kg, at least 0.1 mg / kg, at least 0.2 mg / kg, at least 0.3 mg / kg, at least 0.4 mg / kg, or at least 0.5 mg / kg, and preferably 50 mg / kg or less, 40 mg / kg or less, 30 mg / kg or less, 20 mg / kg or less, or 10 mg / kg or less, for example, about 2.5 mg / day (0.5 mg / kg x 5 kg) to about 5000 mg / day (50 mg / kg x 100 kg). For example, the dosage of the compound can be about 0.1 mg / kg / day to about 50 mg / kg / day, about 0.05 mg / kg / day to about 10 mg / kg / day, about 0.05 mg / kg / day to about 5 mg / kg / day, about 0.05 mg / kg / day to about 3 mg / kg / day, about 0.07 mg / kg / day to about 3 mg / kg / day, about 0.09 mg / kg / day to about 3 mg / kg / day, about 0.05 mg / kg / day to about 0.1 mg / kg / day, about 0.1 mg / kg / day to about 1 mg / kg / day, about 1 mg / kg / day to about 10 mg / kg / day, about 1 mg / kg / day to about 5 mg / kg / day, about 1 mg / kg / day to about 3 mg / kg / day, about 3 mg / day to about 500 mg / day, about 5 mg / day to about 250 mg / day, about 10 mg / day to about 100 mg / day, about 3 mg / day to about 10 mg / day or about 100 mg / day to about 250 mg / day. Such doses can be administered in a single dose or divided into multiple doses. [] [use] [KRAS G12C] [Inhibitor Method] []

[0075] The present disclosure provides methods for inhibiting RAS-mediated cellular signaling, comprising contacting a cell with an effective amount of one or more compounds disclosed herein. Inhibition of RAS-mediated signaling can be assessed and confirmed by a variety of methods known in the art. Non-limiting examples include demonstrating: (a) a decrease in RAS GTPase activity; (b) a decrease in GTP binding affinity or an increase in GDP binding affinity; (c) an increase in K2 dissociation of GTP or a decrease in K2 dissociation of GDP; (d) a decrease in the levels of downstream signaling molecules in the RAS pathway, such as pMEK, pERK, or pAKT; and / or (e) a decrease in the binding of the RAS complex to downstream signaling molecules, including but not limited to Raf. Kits and commercially available assays can be used to determine one or more of the above.

[0076] The present disclosure also provides methods of using the compounds or pharmaceutical compositions of the present disclosure to treat disease conditions, including but not limited to conditions (e.g., cancer) affected by G12C KRAS, HRAS, or NRAS mutations.

[0077] In some embodiments, a method of treating cancer is provided, comprising administering to a subject in need thereof an effective amount of any of the aforementioned pharmaceutical compositions comprising a compound as disclosed herein. In some embodiments, the cancer is mediated by a KRAS, HRAS, or NRAS G12C mutation. In various embodiments, the cancer is pancreatic cancer, colorectal cancer, or lung cancer. In some embodiments, the cancer is gallbladder cancer, thyroid cancer, or bile duct cancer.

[0078] In some embodiments, the present disclosure provides a method of treating a disorder in a subject in need thereof, wherein the method comprises determining whether the subject has a KRAS, HRAS, or NRAS G12C mutation, and if the subject is determined to have a KRAS, HRAS, or NRAS G12C mutation, administering to the subject a therapeutically effective dose of at least one compound as disclosed herein, or a pharmaceutically acceptable salt thereof.

[0079] The disclosed compounds inhibit anchorage-independent cell growth and thus have the potential to inhibit tumor metastasis. Therefore, another embodiment of the present disclosure provides a method for inhibiting tumor metastasis, the method comprising administering an effective amount of a compound disclosed herein.

[0080] KRAS, HRAS, or NRAS G12C mutations have also been identified in hematological malignancies (e.g., cancers affecting the blood, bone marrow, and / or lymph nodes). Therefore, certain embodiments relate to administering the disclosed compounds (e.g., in the form of pharmaceutical compositions) to patients in need of treatment for hematological malignancies. Such malignancies include, but are not limited to, leukemias and lymphomas. For example, the presently disclosed compounds can be used to treat diseases such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myeloid leukemia (CML), acute monocytic leukemia (AMoL), and / or other leukemias. In other embodiments, the compounds can be used to treat lymphomas, such as all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphoma. In various embodiments, the compounds can be used to treat plasma cell malignancies, such as multiple myeloma, pericyte lymphoma, and Waldenstrom's macroglobulinemia.

[0081] Determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can be performed by evaluating the nucleotide sequence encoding the KRAS, HRAS, or NRAS protein, by evaluating the amino acid sequence of the KRAS, HRAS, or NRAS protein, or by evaluating the characteristics of a putative KRAS, HRAS, or NRAS mutant protein. The sequence of wild-type human KRAS, HRAS, or NRAS is known in the art (e.g., Accession No. NP203524).

[0082] Methods for detecting mutations in KRAS, HRAS, or NRAS nucleotide sequences are known to those skilled in the art. Such methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) assays, real-time PCR assays, PCR sequencing, mutant allele-specific PCR amplification (MASA) assays, direct sequencing, primer extension reactions, electrophoresis, oligonucleotide ligation assays, hybridization assays, TaqMan assays, SNP genotyping assays, high-resolution melting assays, and microarray analysis. In some embodiments, samples are evaluated for the G12C KRAS, HRAS, or NRAS mutation by real-time PCR. In real-time PCR, a fluorescent probe specific for the KRAS, HRAS, or NRAS G12C mutation is used. In the presence of the mutation, the probe binds and fluorescence is detected. In some embodiments, a KRAS, HRAS, or NRAS G12C mutation is identified using a direct sequencing method of a specific region in the KRAS, HRAS, or NRAS gene (e.g., exon 2 and / or exon 3). This technique will identify all possible mutations in the sequenced region.

[0083] Methods for detecting mutations in KRAS, HRAS, or NRAS proteins are known to those skilled in the art. Such methods include, but are not limited to, detecting KRAS, HRAS, or NRAS mutants using binding agents (e.g., antibodies) specific for the mutant protein, protein electrophoresis and Western blotting, and direct peptide sequencing.

[0084] Methods for determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can utilize a variety of samples. In some embodiments, the sample is obtained from a subject with a tumor or cancer. In some embodiments, the sample is a fresh tumor / cancer sample. In some embodiments, the sample is a frozen tumor / cancer sample. In some embodiments, the sample is a formalin-fixed, paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

[0085] The present disclosure also relates to a method for treating a hyperproliferative disorder in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound as disclosed herein or a pharmaceutically acceptable salt thereof. In some embodiments, the method relates to treating a subject having cancer, such as acute myeloid leukemia, juvenile cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical teratoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem cell glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoma, embryonal tumor, blastoma, primary lymphoma, cervical cancer, childhood cancer, chordoma, heart tumor, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia, Chronic myeloproliferative leukemia (CML), chronic myeloproliferative disorder, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumor, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal glioma, Ewing sarcoma, extracranial ectodermal tumor, gonadal ectodermal tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), blastoma, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic neuroendocrine tumor Endocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous neck cancer with occult primary, midline cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasms, mycosis fungoides, myelodysplastic syndrome, myeloid metaplasia / myeloproliferative neoplasms, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), mouth cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer In some embodiments, the method is directed to treating a non-cancerous hyperproliferative disorder, such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hypertrophy (BPH)).

[0086] In some embodiments, the methods of treatment relate to treating lung cancer, comprising administering an effective amount of any of the compounds described above (or a pharmaceutical composition comprising such a compound) to a subject in need thereof. In certain embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer, or large cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. Other lung cancers treatable with the disclosed compounds include, but are not limited to, adenomas, carcinoid tumors, and undifferentiated carcinomas.

[0087] The present disclosure further provides methods for modulating the activity of a G12C mutant KRAS, HRAS, or NRAS protein by contacting the protein with an effective amount of a compound of the present disclosure. Modulation can be inhibition or activation of protein activity. In some embodiments, the present disclosure provides methods for inhibiting protein activity by contacting a G12C mutant KRAS, HRAS, or NRAS protein with an effective amount of a compound of the present disclosure in solution. In some embodiments, the present disclosure provides methods for inhibiting the activity of a G12C mutant KRAS, HRAS, or NRAS protein by contacting a cell, tissue, or organ expressing the protein of interest. In some embodiments, the present disclosure provides methods for inhibiting protein activity in a subject, including but not limited to rodents and mammals (e.g., humans), by administering to the subject an effective amount of a compound of the present disclosure. In some embodiments, the percentage of modulation exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some embodiments, the percent inhibition is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0088] In some embodiments, the present disclosure provides methods for inhibiting KRAS, HRAS, or NRAS G12C activity in a cell by contacting the cell with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the cell. In some embodiments, the present disclosure provides methods for inhibiting KRAS, HRAS, or NRAS G12C activity in a tissue by contacting the tissue with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the tissue. In some embodiments, the present disclosure provides methods for inhibiting KRAS, HRAS, or NRAS G12C activity in an organism by contacting the organism with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the organism. In some embodiments, the present disclosure provides methods for inhibiting KRAS, HRAS, or NRAS G12C activity in an animal by contacting the animal with an amount of a compound of the present disclosure sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the animal. In some embodiments, the present disclosure provides methods for inhibiting KRAS, HRAS, or NRAS G12C activity in a mammal by contacting the mammal with an amount of a compound of the present disclosure sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the mammal. In some embodiments, the present disclosure provides methods for inhibiting KRAS, HRAS, or NRAS G12C activity in a human by contacting the human with an amount of a compound of the present disclosure sufficient to inhibit KRAS, HRAS, or NRAS G12C activity in the human. The present disclosure provides methods for treating diseases mediated by KRAS, HRAS, or NRAS G12C activity in a subject in need of such treatment. [] [Combination therapy]

[0089] The present disclosure also provides methods for combination therapy, in which agents known to modulate other pathways or other components of the same pathway, or even overlapping groups of target enzymes, are used in combination with the compounds of the present disclosure or pharmaceutically acceptable salts thereof. In one aspect, such therapy includes, but is not limited to, combining one or more compounds of the present disclosure with chemotherapeutic agents, therapeutic antibodies, and radiation therapy to provide a synergistic or additive therapeutic effect.

[0090] Many chemotherapeutic agents are currently known in the art and can be used in combination with the compounds disclosed herein. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules, such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), Venclexta™ (venatoclax), and Adriamycin™ (doxorubicin), as well as various chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan™); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquinone, meturedopa, and uredopa; ethyleneimines and methylamelamines including hexamethylmelamine, triethylmelamine, triethylphosphatamide, triethylenethiophosphaoramide, and trimethylolomelamine; nitrogen mustards such as chlorambucil, naphthyl mustard, chlorambucil, estramustine, ifosfamide, mechlorethamine oxide, and mechlorethamine hydrochloride; hydrochloride), melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine;Antibiotics, such as aclacinomycin, actinomycin, authramycin, diazoserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, Casodex™, chromomycin, dactinomycin, daunomycin, detoxib, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogamycin, olivomycin, peplomycin, porphyromycin, tfiromycin), puromycin, quelamycin, rhodorubicin, streptozocin, streptozocin, tuberculin, ubenimex, zoloft, doxycycline; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamidine, thioguanine; pyrimidine analogs such as azacitidine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, dooxyfluridine, enocitabine, floxuridine; androgens such as captestosterone, dromostanolone propionate propionate), cyclothioandrostol, melastane, testolactone; antiadrenal drugs such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as frolinic acid; aceglucuronolide; aldophosphamide glycoside; aminoacetyl propionic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; colcemid; diaziquone; elfomithine; elliptonium acetate; etoglucose; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidarol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid acid);2-ethylhydrazine;procarbazine;PSK;propylimine;sizolan;germanylspiramine;alternaria acid;triimidoquinone;2,2',2''-trichlorotriethylamine;urethane;vindesine;dacarbazine;mannomustine;dibromomannitol;dibromodulcitol;pipobroman;gacytosine;arabinoside (Ara-C);Cyclophosphamide; thiotepa; taxanes, such as paclitaxel and docetaxel; retinoic acid; esperamicin; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

[0091] Suitable chemotherapeutic cell conditioning agents also include antihormonal agents used to modulate or inhibit the effects of hormones on tumors, such as antiestrogens, including, for example, tamoxifen (Nolvadex™), raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone and toremifene (Falenton); and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunorubicin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); the topoisomerase inhibitor RFS 2000; and difluoromethylornithine (DMFO).

[0092] If desired, the compounds or pharmaceutical compositions of the present disclosure can be used in combination with commonly prescribed anticancer drugs, such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE, Abagovomab, Acridine carboxamide, Adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, Alpharadin, Alvocidib, 3-aminopyridine-2-carboxaldehyde aminothiourea, Amonafide, Anthracenedione, anti-CD22 immunotoxins, antitumor drugs, antitumorigenic herbs. herbs), Apaziquone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW 2992, Biricodar, Brostallicin, Bryostatin, Buthioninesulfoximine), CBV (chemotherapy), Calyculin, Cell cycle nonspecific antineoplastic agents, Dichloroacetic acid, Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin, Everolimus, Exatecan, Exisulind, Ferruginol, Forodesine, Fosfestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod, Indolocarbazole, Irov Irofulven, Laniquidar, Larotaxel, Lenalidomide, Lucanthone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Ortataxel, PAC-1, Pawpaw, Pixantrone, Proteasome Inhibitors, Rebeccamycin, Resiquimod, Rubitecan, SN-38, Salinosporamide A A), sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126, or zosuquidar.

[0093] The present disclosure further relates to methods for inhibiting abnormal cell growth or treating a hyperproliferative disorder in a mammal using a combination of a compound or pharmaceutical composition provided herein and radiation therapy. Techniques for administering radiation therapy are known in the art and can be used in the combination therapies described herein. Administration of a compound of the present disclosure in such a combination therapy can be determined as described herein.

[0094] Radiation therapy can be administered by any one or a combination of methods, including but not limited to external beam therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy, and permanent or temporary interstitial brachytherapy. As used herein, the term "brachytherapy" refers to radiation therapy delivered by spatially confined radioactive material that is inserted into the body at or near the site of a tumor or other proliferative tissue disease. The term is intended to include, without limitation, exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioisotopes of Lu). Suitable radioactive sources for use as cell conditioning agents of the present disclosure include solids and liquids. By way of non-limiting example, the radiation source can be a radionuclide such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation, or other therapeutic radiation. The radioactive material can also be a fluid prepared from any solution of one or more radionuclides (e.g., a solution of I-125 or I-131), or the radioactive fluid can be produced using a slurry of a suitable fluid containing small particles of solid radionuclides (e.g., Au-198, Y-90). In addition, one or more radionuclides can be embedded in a gel or radioactive microspheres.

[0095] The compounds or pharmaceutical compositions disclosed herein can be used in combination with a certain amount of one or more substances selected from the following: anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors.

[0096] Anti-angiogenic agents, such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors, can be used in conjunction with the compounds of the present disclosure and the pharmaceutical compositions described herein. Examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO 96 / 33172, WO 96 / 27583, European Patent Publication EP 0 818 442, European Patent Publication EP 1 004 578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, WO 98 / 34915, WO 98 / 33768, WO 98 / 30566, European Patent Publication 606 046, European Patent Publication 931 788, WO 90 / 05719, WO 99 / 52910, WO 99 / 52889, WO 99 / 29667, WO 1999007675, European Patent Publication EP 1 786 785, European Patent Publication No. EP 1 181017, U.S. Patent Publication No. US 4845588. 20090012085, U.S. Patent Publication No. 5,863,949, U.S. Patent Publication No. 5,861,510, and European Patent Publication No. 0,780,386, all of which are incorporated herein by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and / or MMP-9 relative to other matrix metalloproteinases (i.e., MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specific examples of MMP inhibitors useful in the present disclosure are AG-3340, RO 32-3555, and RS 13-0830.

[0097] The compounds of the present invention may also be used in co-therapy with other antineoplastic agents, such as acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, hexamethylmelamine, amifostine, aminoacetyl propionic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM 002 (Novelos), bexarotene, bicalutamide, bromuridine, capecitabine, cimothiazine, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030 (Dong-A), daclizumab, denileukin, diftitox), deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon-alpha, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, etifurfural, epirubicin, erythropoietin beta, etoposide phosphate, exemestane, exisulind, fadrozoles, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin), gimeracil / oteracil / tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronic acid, idarubicin, imiquimod, interferon-alpha, interferon-alpha, natural interferon-alpha-2, interferon-alpha-2a, interferon-alpha-2b, interferon-alpha-N1, interferon-alpha-N3, interferon alfacon-1, interferon-alpha, natural interferon-beta, interferon-beta-1a, interferon-beta-1b, interferon-gamma, natural interferon-gamma-1a, interferon-gamma-1b, interleukin-1beta, iobenzylguanidine, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, interferon alpha, leuprorelin, levamisole +Fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprofen, melarsoprol, metoclopramide, mifepristone, miltefosine, milistostim, mismatched double-stranded RNA, mitoguanidine, dibromodulcitol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis-stimulating protein, NSC 631570, octreotide, oprelvekin, osatetron, oxaliplatin, paclitaxel, pamidronate, pegaspargase, pegylated interferon-α-2b, pentosan polysulfate sodium), pentostatin, picibanil, pirarubicin, rabbit anti-thymocyte polyclonal antibody, pegylated interferon-α-2a, porfimer sodium, raloxifene, raltitrexed, rasburiembodiment, rhenium etidronate (Re 186), RII retinamide, rituximab, romotide, samarium (153 Sm) lexidrone lexidronam), sargramostim, sizoran, sobuzosane, sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropin alfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor alpha, natural ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulizin, zinostatin stimalamer or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotides, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diazocone, EL532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocibine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAbs (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotypic 105AD7 MAb (CRC Technology), idiotypic CEA MAb (Trilex), LYM-1-iodine-131 MAb (Techniclone), polymorphic epithelial mucin-yttrium-90 MAb (Antisoma), marimastat, menolipin, mitumomab, motexafin gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), lupinotecan, satraplatin, sodium phenylacetate, sparrosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN, now Pfizer, Inc.), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyl etiopurpurin, tirapazamine, cancer vaccine (Bamira), melanoma vaccine (New York University)University), melanoma vaccine (Sloan Kettering Institute), melanoma tumor lysate vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or valspodar.

[0098] The compounds disclosed herein can further be used together with VEGFR inhibitors. Other compounds described in the following patents and patent applications may be used in combination therapy: US 6,258,812, US 2003 / 0105091, WO 01 / 37820, US 6,235,764, WO 01 / 32651, US 6,630,500, US 6,515,004, US 6,713,485, US 5,521,184, US 5,770,599, US 5,747,498, WO 02 / 68406, WO 02 / 66470, WO 02 / 55501, WO 04 / 05279, WO 04 / 07481, WO 04 / 07458, WO 04 / 09784, WO 02 / 59110, WO 99 / 45009, WO 00 / 59509, WO 99 / 61422, US 5,990,141, WO 00 / 12089 and WO 00 / 02871.

[0099] In some embodiments, the combination comprises a composition of the present disclosure in combination with at least one anti-angiogenic agent. Agents include, but are not limited to, synthetically prepared in vitro chemical compositions, antibodies, antigen-binding regions, radionuclides, and combinations and conjugates thereof. Agents can be agonists, antagonists, allosteric modulators, toxins, or, more generally, can be used to inhibit or stimulate their target (e.g., receptor or enzyme activation or inhibition), thereby promoting cell death or arresting cell growth.

[0100] Exemplary anti-angiogenic agents include ERBITUX™ (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., antibodies or antigen-binding regions that specifically bind to VEGF, or a soluble VEGF receptor or its ligand-binding region) (e.g., AVASTIN™ or VEGF-TRAP™) and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind thereto), EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto) (e.g., Vectibix) panitumumab), IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-Ang1 agents and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind thereto or to their receptors (e.g., Tie2 / Tek)), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto). The pharmaceutical composition disclosed herein may also include one or more agents (e.g., antibodies, antigen binding regions, or soluble receptors) that specifically bind to growth factors and inhibit the activity of the growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor), and antibodies or antigen binding regions that specifically bind to its receptor "c-met".

[0101] Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., U.S. Publication No. 2003 / 0162712; U.S. Patent No. 6,413,932), anti-TWEAK agents (e.g., specific binding antibodies or antigen binding regions, or soluble TWEAK receptor antagonists; see Wiley, U.S. Patent No. 6,727,225), ADAM disintegrin domains for antagonizing the binding of integrins to their ligands (Fanslow et al., U.S. Publication No. 2002 / 0042368), antibodies or antigen binding regions that specifically bind to eph receptors and / or ephrins (U.S. Patent Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124 and members of their patent families), and anti-PDGF-BB antagonists (e.g., antibodies or antigen binding regions that specifically bind to PDGF-BB ligands), and antibodies or antigen binding regions that specifically bind to PDGF-BB ligands and PDGFR kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind thereto).

[0102] Other antiangiogenic / antitumor agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, US 5712291); ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783); vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA, US 5892112); USA), now known as CASI Pharmaceuticals; TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); α-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands); DAC: antiangiogenic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP 970070);ARGENT technology (Ariad, USA);YIGSR-Stealth (Johnson & Johnson, USA);Fibrinogen E fragment (BioActa, UK);Angiogenesis inhibitor (Trigen, UK);TBC-1635 (Encysive Pharmaceuticals, USA);SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); metastasis (Entel, USA); angiogenesis inhibitor (Tripep, Sweden); maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); fluazifop (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN) (Genentech, USA); angiogenesis inhibitor (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); MAb, α5β3 integrin, second generation (Applied Molecular Evolution, USA and MedImmune, USA); gene therapy, retinal disease (Oxford BioMedica, UK); enzastaurin hydrochloride (USAN) (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Cancer Institute, Italy). Institute of Cancer Research, Italy); angiogenesis inhibitors (Alchemia, Australia); VEGF antagonists (Regeneron, USA); rBPI 21 and BPI-derived anti-angiogenic agents (XOMA, USA); PI 88 (Progen, Australia);Cilengitide (pINN) (Merck, Germany; Munich Technical University, Germany; Scripps Clinic and Research Foundation, USA); Cetuximab (INN) (Aventis, France); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292 (Telios, USA); Endostatin (Boston Children's Hospital, USA); ATN 161 (Attenuon, USA); Angiostatin (Boston Children's Hospital, USA); 2-Methoxyestradiol (Boston Children's Hospital, USA); ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK). Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); vatalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); tissue factor pathway inhibitor (Entel, USA); pegaptanib (pINN) (Gilead Sciences, USA); zeocinol (Yonsei University, South Korea); gene-based VEGF-2 vaccine (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX 103 (University of California at San Diego, USA); PX 478 (ProlX, USA) USA); transferstatin (Entel Corporation, USA, now known as CASI Pharmaceuticals); troponin I (Harvard University, USA); SU 6668 (Sugen Corporation, USA, now known as Pfizer Pharmaceuticals);OXI 4503 (OXiGENE, USA); orthoguanidine (Dimensional Pharmaceuticals, USA); motuporamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); atimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); angiogenesis vaccine (Entel, now CASI Pharmaceuticals, USA); urokinase plasminogen activator inhibitor (Dendreon, USA); oglufanide (pINN) (Melmotte, USA) USA); HIF-1α inhibitor (Xenova, UK); CEP 5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); Angisidin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korea Research Institute of Chemical Technology, South Korea); GW 2286 (GlaxoSmithKline, UK); EHT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan) Japan); intraocular 2-methoxyestradiol drug delivery system (Entel, USA); picolinate (anginex) (Maastricht University, Netherlands and University of Minnesota, USA); ABT 510 (Abbott Laboratories, USA); AAL 993 (Novartis, Switzerland);VEGI (ProteomTech, USA); tumor necrosis factor-α inhibitor (National Institute on Aging, USA); SU 11248 (Pfizer and Sugen, USA); ABT 518 (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital and Entel, USA); MAb, KDR (ImClone Systems, USA); MAb, α5β1 (Protein Design, USA); KDR kinase inhibitor (Hiltech, UK and Johnson & Johnson, USA); GFB 116 (South Florida University, USA and Yale University, USA); CS 706 (Sankyo, Japan); Compretin A4 prodrug (Arizona State University, USA). USA); Chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda Company, Japan, and TAP, USA); AG 13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); MAb, vascular endothelial growth factor (Nova Pharmaceuticals, UK); Isoladine (INN) (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany) Germany); squalamine (pINN) (Genaera, USA); RPI 4610 (Sirna, USA);Cancer therapy (Marinova, Australia); heparanase inhibitor (InSight, Israel); KL 3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis AG, Switzerland and Schering AG, Germany); XMP 300 (Xiao Ma, USA); VGA 1102 (Taisho, Japan); VEGF receptor modulator (Pharmacopeia, USA); VE-cadherin-2 antagonist (Bio-Systems, USA); angiostatin (National Institutes of Health, USA); Flk-1 vaccine (Bio-Systems, USA); TZ 93 (Tsumura Co., Ltd., Japan); TumStatin (Beth Israel Hospital, USA); Truncated soluble FLT1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand (Regeneron Pharmaceuticals, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA).

[0103] Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazolecarboxamide ribonucleoside (AICAR), okadaic acid, autophagy-inhibiting algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that increase cAMP levels, such as adenosine, LY204002, N6-mercaptopurine ribonucleoside, and vinblastine. Additionally, antisense or siRNAs that inhibit the expression of proteins, including, but not limited to, ATG5 (which is involved in autophagy), can be used.

[0104] Other pharmaceutically active compounds / agents that can be used to treat cancer and can be used in combination with one or more compounds of the present disclosure include: erythropoietin alfa; darbepoetin alfa; panitumumab; pegfilgrastim; palifermin; filgrastim; denosumab; anciplostim; AMG 102; AMG 176; AMG 386; AMG 479; AMG 655; AMG 745; AMG 951; and AMG 706, or pharmaceutically acceptable salts thereof. []

[0105] In certain embodiments, the compositions provided herein are administered in combination with a chemotherapeutic agent. Suitable chemotherapeutic agents can include natural products such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), taxol, epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunomycin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparaginase, which systemically metabolizes L-asparagine and deprive cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative / antimitotic alkylating agents (e.g., nitrogen mustards, e.g., dichloromethyldiethylamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimines and methylmelamines (e.g., hexamethylmelaamine and thiotepa), CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-03 32991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs and streptozotocin), trazenes-dacarbazinine (DTIC), antiproliferative / antimitotic antimetabolites such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., dapoxetine), (e.g., fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apiracetam, suberoylanilide hydroxamic acid, and dapoxetine). acid), vorinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP (Eg5) inhibitors (e.g., Array 520), DNA binders (e.g., Zalypsis), PI3Kδ inhibitors (e.g., GS-1101 and TGR-1202), PI3Kδ and γ inhibitors (e.g., CAL-130), multikinase inhibitors (e.g.,TG02 and sorafenib), hormones (e.g., estrogens) and hormone agonists such as luteinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprorelin and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38 MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN 163L), Aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CS1 (e.g., elotuzumab), HSP90 inhibitors (e.g., 17 AAG and KOS 953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., Enzastaurin), FTIs (e.g., Zarnestra™), anti-CD138 (e.g., BT062), Torc1 / 2 specific kinase inhibitors (e.g., INK128), kinase inhibitors (e.g., GS-1101), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), BCL-2 antagonists. Other chemotherapeutic agents may include dichloromethane, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or any analog or derivative variant of the foregoing.

[0106] The compounds of the present disclosure may also be used in combination with radiation therapy, hormone therapy, surgery, and immunotherapy, which are well known to those skilled in the art. []

[0107] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with a steroid. Suitable steroids may include, but are not limited to, 21-acetyloxypregnenolone, alclomethasone, algestrel, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazole, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difluprednate, glycyrrhetinic acid, fluzacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinolone acetonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, and fluperolone acetonide. acetate), fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, hydrocortisone, loteprednol etabonate, maprednione, medrysone, methylprednisolone, methylprednisolone, mometasone furoate The compounds disclosed herein include, but are not limited to, furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and their salts and / or derivatives. In a specific embodiment, the compounds disclosed herein can also be used in combination with other pharmaceutically active agents for treating nausea.Examples of agents that can be used to treat nausea include: dronabinol; granisetron; metoclopramide; ondansetron; and prochlorperazine; or pharmaceutically acceptable salts thereof.

[0108] The compounds disclosed herein may also be used in combination with other pharmaceutically active compounds that disrupt or inhibit the RAS-RAF-ERK or PI3K-AKT-TOR signaling pathways. In other such combinations, the other pharmaceutically active compound is a PD-1 and PD-L1 antagonist. The compounds or pharmaceutical compositions disclosed herein may also be used in combination with a certain amount of one or more substances selected from the group consisting of: EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, Mcl-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors, and immunotherapies, including monoclonal antibodies, immunomodulatory imides (IMiDs), anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40 agents, GITR agonists, CAR-T cells, and BiTEs.

[0109] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNA. Useful EGFR antibody inhibitors include cetuximab (Erbitux), panitumumab (Victoza), zalutumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and more recently, lapatinib (TykerB). See, e.g., Yan L et al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005;39(4):565-8 and Paez JG et al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004;304(5676):1497-500.

[0110] Non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications, and all pharmaceutically acceptable salts and solvates of the EGFR inhibitors: European Patent Application EP 520722, published December 30, 1992; European Patent Application EP 566226, published October 20, 1993; PCT International Publication WO 96 / 33980, published October 31, 1996; U.S. Patent No. 5,747,498, issued May 5, 1998; PCT International Publication WO 96 / 30347, published October 3, 1996; European Patent Application EP 787772, published August 6, 1997; PCT International Publication WO 97 / 30034, published August 21, 1997; PCT International Publication WO 97 / 30044, published on August 21, 1997; PCT International Publication WO 97 / 38994, published on October 23, 1997; PCT International Publication WO 97 / 49688, published on December 31, 1997; European Patent Application EP 837063, published on April 22, 1998; PCT International Publication WO 98 / 02434, published on January 22, 1998; PCT International Publication WO 97 / 38983, published on October 23, 1997; PCT International Publication WO 95 / 19774, published on July 27, 1995; PCT International Publication WO 95 / 19970, published on July 27, 1995; PCT International Publication WO 97 / 13771, published April 17, 1997; PCT International Publication WO 98 / 02437, published January 22, 1998; PCT International Publication WO 98 / 02438, published January 22, 1998; PCT International Publication WO 97 / 32881, published September 12, 1997; German Application DE 19629652, published January 29, 1998; PCT International Publication WO 98 / 33798, published August 6, 1998; PCT International Publication WO 97 / 32880, published September 12, 1997; PCT International Publication WO 97 / 32880, published September 12, 1997; European Patent Application EP 682027, published November 15, 1995; PCT International Publication WO 97 / 02266, published on January 23, 1997; PCT International Publication WO 97 / 27199, published on July 31, 1997; PCT International Publication WO 98 / 07726, published on February 26, 1998; PCT International Publication WO 97 / 34895, published on September 25, 1997;PCT International Publication WO 96 / 31510′, published on October 10, 1996; PCT International Publication WO 98 / 14449, published on April 9, 1998; PCT International Publication WO 98 / 14450, published on April 9, 1998; PCT International Publication WO 98 / 14451, published on April 9, 1998; PCT International Publication WO 95 / 09847, published on April 13, 1995; PCT International Publication WO 97 / 19065, published on May 29, 1997; PCT International Publication WO 98 / 17662, published April 30, 1998; U.S. Patent No. 5,789,427, issued August 4, 1998; U.S. Patent No. 5,650,415, issued July 22, 1997; U.S. Patent No. 5,656,643, issued August 12, 1997; PCT International Publication WO 99 / 35146, published July 15, 1999; PCT International Publication WO 99 / 35132, published July 15, 1999; PCT International Publication WO 99 / 07701, published February 18, 1999; and PCT International Publication WO 92 / 20642, published November 26, 1992. Other non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8 (12): 1599-1625.

[0111] Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block activation of EGFR by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi, H. et al., 1993, Br. J. Cancer [British Journal of Cancer] 67: 247-253; Teramoto, T. et al., 1996, Cancer [Cancer] 77: 639-645; Goldstein et al., 1995, Clin. Cancer Res. [Clinical Cancer Research] 1: 1311-1318; Huang, SM et al., 1999, Cancer Res. [Cancer Research] 15: 59 (8): 1935-40; and Yang, X. et al., 1999, Cancer Res. [Cancer Research] 59: 1236-1243. Therefore, the EGFR inhibitor can be the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra), or Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment having the binding specificity thereof.

[0112] The KRAS G12C inhibitors disclosed herein can be used in combination with a MEK inhibitor. Specific MEK inhibitors that can be used in the combinations disclosed herein include PD-325901, trametinib, pimatitinib, MEK162 (also known as binitinib), TAK-733, GDC-0973, and AZD8330. A specific MEK inhibitor that can be used with a KRAS G12C inhibitor in the combinations disclosed herein is trametinib (trade name: Mekinist®, commercially available from Novartis Pharmaceuticals Corp.). Another specific MEK inhibitor is N-(((2R)-2,3-dihydroxypropyl)oxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide, also known as AMG 1009089, 1009089, or PD-325901. Another specific MEK inhibitor that can be used in the combination of the present disclosure includes cobimetinib. MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, and ARRY-438162. []

[0113] PI3K inhibitors include, but are not limited to, wortmannin, a 17-hydroxywortmannin analog described in WO 06 / 044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperidin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]iodine (also known as GDC 0941 and described in PCT Publication Nos. WO 09 / 036,082 and WO 09 / 055,730), 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT Publication No. WO 09 / 036,082 and WO 09 / 055,730). 06 / 122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-thiolinothieno[3,2-d]pyrimidin-6-yl)methyl)piperidin-1-yl)-2-hydroxypropan-1-one (described in PCT Publication No. WO 2008 / 070740), LY 294002 (2-(4-thiolino)-8-phenyl-4H-1-benzopyran-4-one, available from Axon Medchem), PI 103 hydrochloride (3-[4-(4-thiolinopyrido-[3′,2′:4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride, available from Axon Medchem), PIK 75 (N′-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfonyl-hydrazine hydrochloride, available from Exon Medical Chemicals), PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide, available from Exon Medical Chemicals), GDC-0941 dimethanesulfonate (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperidin-1-ylmethyl)-4-oxo-thieno[3,2-d]pyrimidine, available from Exon Medical Chemicals), AS-252424 ( 5-[1-[5-(4-Fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione, available from Exon Medical Chemicals) and TGX-221 (7-methyl-2-(4-oxo-1,2-dextrin-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one, available from Exon Medical Chemicals), XL-765, and XL-147.Other PI3K inhibitors include demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0114] AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385(Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and 2) (Barnett et al. (2005) Biochem. J., 385(Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05011700); indole-3-carbinol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134 (12 Suppl), 3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; Dasmahapatra et al. (2004) Clin. Cancer Res. 10 (15), 5242-52, 2004); phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); and triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al. (2004) Cancer Res. 64, 4394-9).

[0115] TOR inhibitors include, but are not limited to, AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, and ATP-competitive TORC1 / TORC2 inhibitors, including PI-103, PP242, PP30, and Torin 1. Other TOR inhibitors include FKBP12 enhancers; rapamycin and its derivatives, including: CCI-779 (temsirolimus), RAD001 (everolimus; WO 9409010) and AP23573; rapamycin analogs (rapalogs), such as AP23573, AP23464 or AP23841 as disclosed in WO 98 / 02441 and WO 01 / 14387; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropionate]-rapamycin (also known as CC1779), 40-epi-(tetrazolyl)-rapamycin (also known as ABT578), 32-deoxyrapamycin, 16-pentynyloxy-32(S)-dihydrorapamycin and WO 98 / 02441. 05005434; derivatives disclosed in the following patents: U.S. Patent No. 5,258,389, WO 94 / 090101, WO 92 / 05179, U.S. Patent No. 5,118,677, U.S. Patent No. 5,118,678, U.S. Patent No. 5,100,883, U.S. Patent No. 5,151,413, U.S. Patent No. 5,120,842, WO 93 / 111130, WO 94 / 02136, WO 94 / 02485, WO 95 / 14023, WO 94 / 02136, WO 95 / 16691, WO 96 / 41807, WO 96 / 41807 and U.S. Patent No. 5,256,790; phosphorus-containing rapamycin derivatives (e.g., WO 05016252); 4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional Application No. 60 / 528,340).

[0116] MCl-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the key anti-apoptotic members of the B-cell lymphoma 2 (BCL-2) protein family. Overexpression of MCL-1 is closely associated with tumor progression and resistance not only to traditional chemotherapy but also to targeted therapies including BCL-2 inhibitors (e.g., ABT-263).

[0117] In the present disclosure, KRAS G12C inhibitors can also be used in combination with SHP2 inhibitors. SHP2 inhibitors that can be used in the present combination include, but are not limited to, SHP099, and RMC-4550 or RMC-4630 (available from Revolutions Medicines, Redwood City, CA).

[0118] Proteasome inhibitors include, but are not limited to, Kyprolis® (carfilzomib), Velcade® (bortezomib), and oprozomib.

[0119] Immunotherapy includes, but is not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents.

[0120] Monoclonal antibodies include, but are not limited to, Darzalex® (daratumumab), Herceptin® (trastuzumab), Avastin® (bevacizumab), Rituxan® (rituximab), Lucentis® (ranibizumab), and Eylea® (aflibercept).

[0121] Immunomodulators (IMiDs) are a class of immunomodulatory drugs (drugs that modulate immune responses) that contain an imide group. IMiDs include thalidomide and its analogs (lenalidomide, pomalidomide, and amilast).

[0122] Anti-PD-1 inhibitors, including but not limited to antibodies, include but are not limited to pembrolizumab (Keytruda®), AMG 404, and nivolumab (Opdivo®). Exemplary anti-PD-1 antibodies and methods of use thereof are described in Goldberg et al., Blood 110(1):186-192 (2007); Thompson et al., Clin. Cancer Res. 13(6):1757-1761 (2007); and Korman et al., International Application No. PCT / JP 2006 / 309606 (Publication No. WO 2006 / 121168 A1), each of which is expressly incorporated herein by reference. These include: Yervoy™ (ipilimumab) or Tremelimumab (targeting CTLA-4), galiximab (targeting B7.1), BMS-936558 (targeting PD-1), MK-3475 (targeting PD-1), AMP224 (targeting B7DC), BMS-936559 (targeting B7-H1), MPDL3280A (targeting B7-H1), MEDI-570 (targeting ICOS), AMG557 (targeting B7H2), MGA271 (targeting B7H3), IMP321 (targeting LAG-3), BMS-663513 (targeting CD137), PF-05082566 (targeting CD137), CDX-1127 (targeting CD27), anti-OX40 (Providence Health Services Services), huMAbOX40L (targeting OX40L), Atacicept (targeting TACI), CP-870893 (targeting CD40), Lucatumumab (targeting CD40), Dacetuzumab (targeting CD40), Muromonab-CD3 (targeting CD3), and Ipilimumab (targeting CTLA-4). Immunotherapy also includes genetically engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTEs).

[0123] GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as the GITR fusion proteins described in U.S. Patent No. 6,111,090 box.c, European Patent No. 090505B1, U.S. Patent No. 8,586,023, PCT Publication Nos. WO 2010 / 003118 and 2011 / 090754, or anti-GITR antibodies such as those described in U.S. Patent No. 7,025,962, European Patent No. 1947183B1, U.S. Patent No. 7,812,135, U.S. Patent No. 8,388,967, U.S. Patent No. 8,591,886, European Patent No. EP 1866339, PCT Publication No. WO 2011 / 028683, PCT Publication No. WO 2013 / 039954, PCT Publication No. WO 2005 / 007190, PCT Publication No. WO 2007 / 133822, PCT Publication No. WO 2005 / 055808, PCT Publication No. WO 99 / 40196, PCT Publication No. WO 2001 / 03720, PCT Publication No. WO 99 / 20758, PCT Publication No. WO 2006 / 083289, PCT Publication No. WO 2005 / 115451, U.S. Patent No. 7,618,632 and PCT Publication No. WO 2011 / 051726.

[0124] The compounds described herein may be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated. Thus, in some embodiments, one or more compounds of the present disclosure will be co-administered with other agents as described above. When used in combination therapy, the compounds described herein and the second agent are administered simultaneously or separately. This combined administration can include simultaneous administration of both agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compounds described herein and any of the above-mentioned agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds of the present disclosure and any of the above-mentioned agents can be administered simultaneously, with the two agents being present in separate formulations. In another alternative, any of the above-mentioned agents can be administered immediately after the compounds of the present disclosure, or vice versa. In some embodiments of separate administration regimens, the compounds of the present disclosure and any of the above-mentioned agents are administered a few minutes, a few hours, or a few days apart. []

[0125] While one aspect of the present disclosure contemplates treating diseases / conditions with combinations of pharmaceutically active compounds that can be administered separately, the present disclosure further relates to combining separate pharmaceutical compositions in a kit format. The kit comprises two separate pharmaceutical compositions: a compound of the present disclosure and a second pharmaceutical compound. The kit comprises containers for holding the separate compositions, such as divided bottles or divided foil pouches. Other examples of containers include syringes, boxes, and bags. In some embodiments, the kit comprises instructions for use of the separate components. The kit format is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), at different dosage intervals, or when titration of the individual components of the combination is desired by the prescribing healthcare professional.

[0126] All patents and other publications cited herein are incorporated by reference.

[0127] The methods presented below illustrate specific embodiments of the present disclosure. These methods are representative and are not intended to limit the scope of the claims in any way. [] [Related Synthesis Methods] []

[0128] The following intermediate compounds of 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenyl)-1-piperidinyl)pyrido[2,3-d]pyrimidin-2(1H)-one are representative examples of the present disclosure and should not be construed as limiting the scope of the present invention.

[0129] The synthesis of compound 9 and related intermediates is described in U.S. Serial No. 15 / 984,855, filed May 21, 2018 (U.S. Publication No. 2018 / 0334454, November 22, 2018), which claims priority to and claims the benefit of U.S. Provisional Application No. 62 / 509,629, filed May 22, 2017, both of which are incorporated herein by reference in their entireties for all purposes. 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenyl)-1-piperidinyl)pyrido[2,3-d]pyrimidin-2(1H)-one was prepared using the following method, in which the isomers of the final product were separated by chiral chromatography.

[0130] [step] [1] [:] [2,6-] [Dichloro] [-5-] [Flunicotinamide (intermediate)] [S] To a mixture of 2,6-dichloro-5-fluoro-nicotinic acid (4.0 g, 19.1 mmol, AstaTech Inc., Bristol, Pennsylvania) in dichloromethane (48 mL) was added oxalyl chloride (2M in DCM, 11.9 mL, 23.8 mmol), followed by a catalytic amount of DMF (0.05 mL). The reaction was stirred at room temperature overnight and then concentrated. The residue was dissolved in 1,4-dioxane (48 mL) and cooled to 0°C. Ammonium hydroxide solution (28.0%-30% based on NH3, 3.6 mL, 28.6 mmol) was slowly added via syringe. The resulting mixture was stirred at 0°C for 30 min and then concentrated. The residue was diluted with a 1:1 mixture of EtOAc / heptane and stirred for 5 min before filtering. The filtered solid was discarded, and the remaining mother liquor was concentrated to half its volume and filtered. The filtered solid was washed with heptane and dried in a reduced pressure oven (45°C) overnight to provide 2,6-dichloro-5-fluoronicotinamide. H NMR (400 MHz, DMSO-d 6) δ ppm 8.23 ​​(d, J = 7.9 Hz, 1 H) 8.09 (br s, 1 H) 7.93 (br s, 1 H). m / z (ESI, positive ion): 210.9 (M+H).

[0131] [step] [2] [:] [2,6-] [Dichloro] [-5-] [fluorine] [- , N , -((2- ] [Isopropyl] [-4-] [Methylpyridine] [-3-] [base] [)] [Carbamoyl] [)] [Niacinamide.] Add ice-cold 2,6-dichloro-5-fluoronicotinamide ( [Intermediates] To a slurry of [S], 5.0 g, 23.9 mmol) in THF (20 mL) was slowly added oxalyl chloride (2M in DCM, 14.4 mL, 28.8 mmol). The resulting mixture was heated at 75°C for 1 h, then the heat was removed and the reaction was concentrated to half its volume. After cooling to 0°C, THF (20 mL) was added dropwise via cannula, followed by 2-isopropyl-4-methylpyridin-3-amine ( [Intermediates] [R], 3.59 g, 23.92 mmol) in THF (10 mL). The resulting mixture was stirred at 0°C for 1 h, then quenched with a 1:1 mixture of brine and saturated aqueous ammonium chloride. The mixture was extracted with EtOAc (3x), and the combined organic layers were dried over anhydrous sodium sulfate and concentrated to provide 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)aminoformyl)nicotinamide. This material was used in the following step without further purification. m / z (ESI, +ve ion): 385.1 (M+H) +.

[0132] [step] [3] [:] [7-] [chlorine] [-6-] [fluorine] [-1-(2-] [Isopropyl] [-4-] [Methylpyridine] [-3-] [base] [)] [Pyrido] [[2,3-d]] [Pyrimidine] [-2,4(1H,3H)-] [Diketone.] To an ice-cooled solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)aminoformyl)nicotinamide (9.2 g, 24.0 mmol) in THF (40 mL) was slowly added KHMDS (1 M in THF, 50.2 mL, 50.2 mmol) via syringe. The ice bath was removed, and the resulting mixture was stirred at room temperature for 40 min. The reaction was quenched with saturated aqueous ammonium chloride and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH / heptane) to provide 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione. 1H NMR (400 MHz, DMSO- d 6 ) δ ppm 12.27 (br s, 1H), 8.48-8.55 (m, 2 H), 7.29 (d, J= 4.8 Hz, 1 H), 2.87 (quin, J= 6.6 Hz, 1 H), 1.99-2.06 (m, 3 H), 1.09 (d, J= 6.6 Hz, 3 H), 1.01 (d, J= 6.6 Hz, 3 H). 19F NMR (376 MHz, DMSO-d 6) δ: -126.90 (s, 1 F). m / z (ESI, +ve ion): 349.1 (M+H) +.

[0133] [step] [4] [:] [4,7-] [Dichloro] [-6-] [fluorine] [-1-(2-] [Isopropyl] [-4-] [Methylpyridine] [-3-] [base] [)] [Pyrido] [[2,3- , d , ] ] [Pyrimidine] [-2(1H)-] [Ketone.] Phosphorus oxychloride (1.63 mL, 17.5 mmol) was added dropwise via syringe to a solution of 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (4.7 g, 13.5 mmol) and DIPEA (3.5 mL, 20.2 mmol) in acetonitrile (20 mL). The resulting mixture was heated at 80°C for 1 h, then cooled to room temperature and concentrated to provide 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. This material was used in the following step without further purification. m / z (ESI, +ve ion): 367.1 (M+H)+.

[0134] [step] [5] [:] [4-(7-] [chlorine] [-6-] [fluorine] [-1-(2-] [Isopropyl] [-4-] [Methylpyridine] [-3-] [base] [)-2-] [Pendant Oxygen Group] [-1,2-] [Dihydropyrido] [[2,3- , d , ] ] [Pyrimidine] [-4-] [base] [)-3-] [Methylpiperidin] [𠯤] [-1-] [Formic acid] [( , S , )- ] [Tributyl ester.] To an ice-cold solution of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (13.5 mmol) in acetonitrile (20 mL) was added DIPEA (7.1 mL, 40.3 mmol), followed by (S)-4-N-Boc-2-methylpiperidinium (3.23 g, 16.1 mmol, Combi-Blocks, Inc., San Diego, CA, USA). The resulting mixture was warmed to room temperature and stirred for 1 h, then diluted with cold saturated aqueous sodium bicarbonate (200 mL) and EtOAc (300 mL). The mixture was stirred for an additional 5 min, the layers separated, and the aqueous layer extracted with more EtOAc (1x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0%-50% EtOAc / heptane) to provide 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperidin-1-carboxylic acid (S)- [] Tertiary butyl ester. m / z (ESI, +ve ion): 531.2 (M+H) +.

[0135] [step] [6] [:] [4-(6-] [fluorine] [-7-(2-] [fluorine] [-6-] [Hydroxyphenyl] [)-1-(2-] [Isopropyl] [-4-] [Methylpyridine] [-3-] [base] [)-2-] [Pendant Oxygen Group] [-1,2-] [Dihydropyrido] [[2,3- , d , ] ] [Pyrimidine] [-4-] [base] [)-3-] [Methylpiperidin] [𠯤] [-1-] [Formic acid] [(3 , S , )- ] [Tributyl ester.] 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperidin-1-carboxylic acid (S)- [] tertiary butyl ester (4.3 g, 8.1 mmol), potassium trifluoro(2-fluoro-6-hydroxyphenyl)borate ( [Intermediate] A mixture of [Q] (2.9 g, 10.5 mmol), potassium acetate (3.2 g, 32.4 mmol), and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (661 mg, 0.81 mmol) complexed with dichloromethane in 1,4-dioxane (80 mL) was degassed with nitrogen for 1 minute. Deoxygenated water (14 mL) was added, and the resulting mixture was heated at 90°C for 1 hour. The reaction was cooled to room temperature, quenched with semisaturated aqueous sodium bicarbonate, and extracted with EtOAc (2x) and DCM (1x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-60% 3:1 EtOAc-EtOH / heptane) to provide 4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperidin-1-carboxylic acid (3S)- [] Tertiary butyl ester. 1H NMR(400 MHz, DMSO- d 6 )δ ppm 10.19(br s, 1 H), 8.38(d, J= 5.0 Hz, 1 H), 8.26(dd, J= 12.5, 9.2 Hz, 1 H), 7.23-7.28(m, 1 H), 7.18(d, J= 5.0 Hz, 1 H), 6.72(d, J= 8.0 Hz, 1 H), 6.68(t, J= 8.9 Hz, 1 H), 4.77-4.98(m, 1 H), 4.24(br t, J= 14.2 Hz, 1 H), 3.93-4.08(m, 1 H), 3.84(br d, J= 12.9 Hz, 1 H), 3.52-3.75(m, 1 H), 3.07-3.28(m, 1 H), 2.62-2.74(m, 1 H), 1.86-1.93(m, 3 H), 1.43-1.48(m, 9 H), 1.35(dd, J= 10.8, 6.8 Hz, 3 H), 1.26-1.32(m, 1 H), 1.07(dd, J= 6.6, 1.7 Hz, 3 H), 0.93(dd, J= 6.6, 2.1 Hz, 3 H). 19F NMR (376 MHz, DMSO-d 6) δ: -115.65 (s, 1 F), -128.62 (s, 1 F). m / z(ESI, +ve ion): 607.3 (M+H) +.

[0136] [step] [7] [:] [6-] [fluorine] [-7-(2-] [fluorine] [-6-] [Hydroxyphenyl] [)-1-(4-] [methyl] [-2-(2-] [Propanyl] [)-3-] [Pyridyl] [)-4-((2 , S , )-2- ] [methyl] [-4-(2-] [Acryloyl] [)-1-] [piperidin] [𠯤] [base] [)] [Pyrido] [[2,3- , d , ] ] [Pyrimidine] [-2(1H)-] [Ketone.] Trifluoroacetic acid (25 mL, 324 mmol) was added to a solution of (3S)-tert-butyl 4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperidin-1-carboxylate (6.3 g, 10.4 mmol) in DCM (30 mL). The resulting mixture was stirred at room temperature for 1 h and then concentrated. The residue was dissolved in DCM (30 mL), cooled to 0°C, and treated sequentially with DIPEA (7.3 mL, 41.7 mmol) and acryloyl chloride (0.849 mL, 10.4 mmol) in DCM (3 mL; added dropwise via syringe). The reaction was stirred at 0°C for 10 min, then quenched with half-saturated aqueous sodium bicarbonate solution and extracted with DCM (2x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-100% 3:1 EtOAc-EtOH / heptane) to provide 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenyl)-1-piperidinyl)pyrido[2,3-d]pyrimidin-2(1H)-one. 1H NMR(400 MHz, DMSO- d 6 )δppm 10.20(s, 1 H), 8.39(d, J= 4.8 Hz, 1 H), 8.24-8.34(m, 1 H), 7.23-7.32(m, 1 H), 7.19(d, J= 5.0 Hz, 1 H), 6.87(td, J= 16.3, 11.0 Hz, 1 H), 6.74(d, J= 8.6 Hz, 1 H), 6.69(t, J= 8.6 Hz, 1 H), 6.21(br d, J= 16.2 Hz, 1 H), 5.74-5.80(m, 1 H), 4.91(br s, 1 H), 4.23-4.45(m, 2 H), 3.97-4.21(m, 1 H), 3.44-3.79(m, 2 H), 3.11-3.31(m, 1 H), 2.67-2.77(m, 1 H), 1.91(s, 3 H), 1.35(d, J= 6.8 Hz, 3 H), 1.08(d, J= 6.6 Hz, 3 H), 0.94(d, J= 6.8 Hz, 3 H).19F NMR (376 MHz, DMSO-d 6) δ ppm -115.64 (s, 1 F), -128.63 (s, 1 F). m / z(ESI, +ve ion): 561.2 (M+H) +. 。

[0137] Another synthesis of compound 9 and related intermediates is described in U.S. provisional patent application filed on November 16, 2018, which is incorporated herein by reference in its entirety for all purposes. [Representative Synthesis Methods] []

[0138] The present disclosure includes the following steps, wherein the synthesis and utilization of a boroxine intermediate in the preparation of AMG 510 (Compound 9) are novel and inventive steps. [raw materials] [] [Material] [structure] [CAS #] [MW] [(] [g / mol] [)] (2,6-dichloro-5-fluoronicotinamide) Compound 1 113237-20-0 209.99 2-Isopropyl-4-methylpyridin-3-amine Compound 2A 1698293-93-4 150.22 (S)-1-Boc-3-methylpiperidinium 147081-29-6 200.28 2,2',2''-(1,3,5,2,4,6-trioxatriborane-2,4,6-triyl)tris(3-fluorophenol) Compound 6A N / A 413.71 Acryloyl chloride 814-68-6 90.51 NOTE: In these incoming starting materials, the Des-boc content in the amine and the 3-chloropropionyl chloride content in the acrylyl chloride need to be controlled to ensure adequate final drug substance quality. [] [step] [1a] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [ / ] [volume] [Moer] [theory] 2,6-Dichloro-5-fluoro-3-pyridinecarboxylic acid 82671-06-5 209.99 1.0 equivalent 119.1 25 kg DCM 74-09-2 84.93 16.51 equivalent 2354.9 200 kg DMF 68-12-2 73.09 0.068 equivalent 8.1 592 g (627 mL) Oxaloyl chloride 79-37-8 126.93 1.25 equivalent 148.9 18.9 kg Ammonium hydroxide 1336-21-6 35.05 5 equivalents 595.5 40.2 L water 7732-18-5 18.02 N / A N / A 261 L

[0139] To a solution of 2,6-dichloro-5-fluoro-3-pyridinecarboxylic acid (25 kg; 119.1 mol) in dichloromethane (167 kg) and DMF (592 g) was added oxalyl chloride (18.9 kg; 148.9 mol), while maintaining the internal temperature between 15°C and 20°C. Additional dichloromethane (33 kg) was added as a rinse, and the reaction mixture was stirred for 2 hours. The reaction mixture was cooled and then quenched with ammonium hydroxide (40.2 L; 595.5 mol), while maintaining the internal temperature at 0 ± 10°C. The resulting slurry was stirred for 90 minutes, after which the product was collected by filtration. The filtered solid was washed with deionized water (3 x 87 L) and dried to provide 2,6-dichloro-5-fluoronicotinamide (Compound 1). [] [step] [1b] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [ / ] [volume] [Moer] [theory] amide (2,6-dichloro-5-fluoronicotinamide) 113237-20-0 209.99 1.0 equivalent 77.8 16.27 kg Oxaloyl chloride 79-37-8 126.93 1.2 equivalent 93.8 11.9 kg (7.9 L) dichloromethane 75-09-2 84.93 N / A N / A 730.7 kg (551.5 L) Aniline DCM solution 2-Isopropyl-4-methylpyridin-3-amine 1698293-93-4 150.22 1.1 equivalent 85.9 12.9 kg (including weight of aniline)

[0140] In Reactor A, a solution of 2,6-dichloro-5-fluoronicotinamide (Compound 1) (16.27 kg; 77.8 mol) in dichloromethane (359.5 kg) was added with oxalyl chloride (11.9 kg; 93.8 mol) while maintaining the temperature at ≤ 25°C for 75 minutes. The resulting solution was then heated to 40°C ± 3°C and aged for 3 hours. Using vacuum, the solution was distilled to remove dichloromethane until the solution was below the stirrer. Dichloromethane (300 kg) was then added, and the mixture was cooled to 0 ± 5°C. To a clean, dry reactor (Reactor B), 2-isopropyl-4-methylpyridin-3-amine (Aniline Compound 2A) (12.9 kg; 85.9 mol) was added, followed by dichloromethane (102.6 kg). The aniline solution was azeotropically dried by vacuum distillation while maintaining the internal temperature between 20°C and 25°C, replacing it with additional dichloromethane until the solution was dry (limit ≤ 0.05%) as determined by KF analysis. The volume of the solution was adjusted to approximately 23 L with dichloromethane. The dry aniline solution was then added to Reactor A, maintaining the internal temperature at 0 ± 5°C throughout the addition. The mixture was then heated to 23°C and aged for 1 hour. The solution was finely filtered into a clean reactor to yield a DCM solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)aminoformyl)nicotinamide (Compound 3), which was used directly in the next step. [] [step] [2] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [ / ] [volume] [Moer] [theory] Urea, DCM solution 2,6-Dichloro-5-fluoro-N-{[4-methyl-2-(propan-2-yl)pyridin-3-yl]aminocarbonyl}pyridine-3-carboxamide N / A 385.22 1.0 equivalent 38.9 208.3 kg (including 15 kg weight) 2-Methyltetrahydrofuran 96-47-9 86.13 N / A N / A 308 kg (358 L) Tertiary sodium butoxide 865-48-5 96.11 2.0 equivalent 97.8 9.4 kg Ammonium chloride 12125-02-9 53.49 N / A 430 23.0 kg hydrochloric acid 7467-01-0 36.46 N / A 41 1.6 kg magnesium sulfate 7487-88-9 120.37 N / A 195 23.5 kg Sodium chloride 7647-14-5 58.44 N / A 282 16.5 kg heptane 142-82-5 100.21 N / A N / A 94 L 10% citric acid 75 kg

[0141] A solution of 2,6-dichloro-5-fluoro-N-{[4-methyl-2-(propan-2-yl)pyridin-3-yl]carbamyl}pyridine-3-carboxamide (UREA (Compound 3)) (containing 15 kg; 38.9 mol) in dichloromethane was solvent-exchanged with 2-MeTHF using vacuum distillation while maintaining an internal temperature of 20°C-25°C. The reactor volume was adjusted to 40 L and then charged with additional 2-MeTHF (105.4 kg). Sodium tert-butoxide (9.4 kg; 97.8 mol) was added while maintaining a temperature of 5°C-10°C. The contents were warmed to 23°C and stirred for 3 hours. The contents were then cooled to 0-5°C, and a solution of ammonium chloride (23.0 kg; 430 mol) in 60 L of deionized water was added. The mixture was warmed to 20°C, deionized water (15 L) was added, and further aged for 30 minutes. Stop stirring and separate the layers. Remove the aqueous layer, and add deionized water (81.7 L) to the organic layer. Prepare a mixture of concentrated HCl (1.5 kg) and water (9 L) and slowly add it to the reactor until the measured pH is between 4 and 5. Separate the layers, and back-extract the aqueous layer with 2-MeTHF (42.2 kg). Combine the two organic layers and wash with 10% citric acid solution (75 kg), followed by a mixture of water (81.7 L) and saturated NaCl (19.8 kg). The organic layer is then washed with saturated sodium bicarbonate (75 kg), repeating as necessary to achieve an aqueous target pH ≥ 7.0. Wash the organic layer again with brine (54.7 kg) and then dry with magnesium sulfate (5 kg). Filter the mixture to remove the magnesium sulfate, and rinse the filter bed with 2-MeTHF (49.2 kg). Distill the combined filtrate and washings under vacuum to a volume of 40 L. The concentrated solution was heated to 55°C and heptane (10-12 kg) was slowly added until the cloud point was reached. The solution was cooled to 23°C over 2 hours, and then heptane (27.3 kg) was added over 2 hours. The product slurry was aged at 20-25°C for 3 hours, then filtered and washed with a mixture of 2-MeTHF (2.8 kg) and heptane (9 kg). The product was dried under nitrogen and vacuum to obtain solid 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (racemic-diketone (Compound 4)). [] [step] [3] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [ / ] [volume] [Moer] [theory] Racemic-diketone (Compound 4) N / A 348.76 1.0 (+)-2,3-Diphenylmethane-D-tartaric acid 17026-42-5 358.30 2.0 2-Methyltetrahydrofuran 96-47-9 86.13 7.0 heptane 142-82-5 100.21 2.0 heptane 142-82-5 100.21 3.0 2-Methyltetrahydrofuran 96-47-9 86.13 4.0 heptane 142-82-5 100.21 2.0

[0142] Under a nitrogen atmosphere, (+)-2,3-diphenylcarbamyl-D-tartaric acid (2.0 equiv) was added to a stirred suspension of compound 4 (1.0 equiv) in 2-methyltetrahydrofuran (7.0 L / kg) in a vessel. 2-MeTHF is chiral but was used as a racemic mixture. Different enantiomers of 2-MeTHF were randomly incorporated into the co-crystals. The resulting suspension was warmed to 75°C and aged at 75°C until complete dissolution was observed (≤ 30 min). The resulting solution was finely filtered at 75°C into a second vessel. n-heptane (2.0 L / kg) was added to the finely filtered solution at a rate that maintained the internal temperature above 65°C. The solution was then cooled to 60°C, seeded with crystals (0.01 kg / kg), and aged for 30 minutes. The resulting suspension was cooled to 20°C over 4 hours and then sampled for chiral purity analysis by HPLC. The suspension was charged with n-heptane (3.0 L / kg) and then aged at 20°C for 4 hours under a nitrogen atmosphere. The suspension was filtered, and the separated solid was washed twice with (2:1) n-heptane:2-methyltetrahydrofuran (3.0 L / kg). The material was dried under nitrogen and vacuum to obtain a meta-diketone:DBTA:Me-THF complex (Compound 4a). [] [step] [4] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [ / ] [volume] [Moer] [theory] Meta-diketone / DBTA / Me-THF cocrystal (Compound 4a) N / A 1228.08 1.0 74.2 46.9 kg (25.9 kg corrected for meta-diketone) Methyl tertiary butyl ether 1634-04-4 88.15 45.0 17593 2100 L Disodium hydrogen phosphate 7558-79-4 141.96 2.0 148.4 21.1 kg USP Purified Water As needed magnesium sulfate 7487-88-9 120.37 N / A N / A 25 kg heptane 142-82-5 100.20 60.0 19322 2835 L

[0143] In vessel A, stir a suspension of sodium hydrogen phosphate (21.1 kg, 2.0 equivalents) in deionized water (296.8 L, 6.3 L / kg) until dissolution is observed (≥ 30 min). In vessel B, stir a suspension of meta-diketone: DBTA: Me-THF complex (Composition 4a) [46.9 kg (25.9 kg, 1.0 equivalents corrected for meta-diketone)] in methyl tert-butyl ether (517.8 L, 11.0 L / kg) for 15 to 30 minutes. Add the resulting solution from vessel A to vessel B, and stir the mixture for over 3 hours. Stop stirring, and allow the biphasic mixture to separate for over 30 minutes. Remove the lower aqueous phase, then back-extract with methyl tert-butyl ether (77.7 L, 1.7 L / kg). Combine the organic phases in vessel B and dry over magnesium sulfate (24.8 kg, 0.529 kg / kg). The resulting suspension from container B was stirred for over three hours and then filtered into container C. A rinse of methyl tert-butyl ether (46.9 L, 1.0 L / kg) was added to container B and then filtered into container C. The contents of container C were cooled to 10°C and then distilled under vacuum while slowly warming to 35°C. Distillation was continued until 320-350 kg (6.8-7.5 kg / kg) of methyl tert-butyl ether was collected. After cooling the contents of container C to 20°C, n-heptane (278.7 L, 5.9 L / kg) was added over 1 hour and then distilled under vacuum while slowly warming to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane was collected. After cooling the contents of container C to 20°C, n-heptane (278.7 L, 5.9 L / kg) was added a second time over 1 hour, followed by distillation under vacuum while slowly warming to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane was collected. After cooling the contents of container C to 20°C, n-heptane (195.9 L, 4.2 L / kg) was added a third time over 1 hour, followed by sampling for solvent composition analysis by GC. The suspension in container C was stirred for an additional hour. The suspension was filtered and then washed from container C with a rinse of n-heptane (68.6 L, 1.5 L / kg). The isolated solid was dried at 50°C and a sample was submitted for storage. 7-Chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (meta-diketone) compound 5M was obtained.

[0144] The first-generation method highlighted above has been successfully scaled up to 200+ kg of rac-dione starting material (Compound 4). In this method, seeding with a thermodynamically stable crystalline form of the rac-dione (which exhibits low solubility) resulted in batch failure. Based on our subsequent studies, we discovered that adjusting the heptane feed schedule to increase the DBTA equivalents and lower the seed temperature improved the robustness of the method. The improved method is resistant to the presence of the thermodynamically stable crystalline form of the rac-dione and facilitates successful separation of atropisomers. Subsequent batches will incorporate the improved method for large-scale manufacturing. [] [step] [5] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [enter] Meta-diketone 5M N / A 348.76 1 equivalent 1 equivalent Toluene-1 108-88-3 92.14 10.0 L / kg Toluene-2 108-88-3 92.14 0.5 L / kg Toluene-3 108-88-3 92.14 4.5 L / kg Phosphate chloride 10025-87-3 153.33 1.5 equivalent N,N-Diisopropylethylamine-1 7087-68-5 129.24 2.0 equivalent N,N-Diisopropylethylamine-2-yl 7087-68-5 129.24 1.2 equivalent (S)-1-Boc-3-methylpiperidinium 147081-29-6 200.28 1.2 equivalent Sodium bicarbonate 144-55-8 84.01 4.5 equivalent Water-1 18.01 15.0 L / kg dichloromethane 75-09-2 84.93 1.0 L / kg Isopropyl acetate-1 108-21-4 102.132 1.0 L / kg Water-2 18.01 5.0 L / kg Water-3 18.01 5.0 L / kg Isopropyl acetate-2 108-21-4 102.132 5.0 L / kg Isopropyl acetate-3 108-21-4 102.132 5.0 L / kg acetone 67-64-1 58.08 10.0 L / kg Water-4 18.01 10.0 L / kg 1:1 acetone / water N / A N / A 5.0 L / kg Note: All L / kg amounts are relative to meta-dionone input; all equivalent amounts are relative to meta-dionone input adjusted for potency.

[0145] Meta-diketone (Compound 5M, 1.0 equivalent) and toluene-1 (10.0 L / kg) were charged to vessel A. The resulting solution was dried by azeotropic distillation under vacuum at 45°C until 5.0 L / kg of solvent had been removed. The contents of vessel A were then cooled to 20°C.

[0146] Vessel C was filled with toluene-3 (4.5 L / kg), phosphatidyl chloride (1.5 eq.), and N,N-diisopropylethylamine-1 (2.0 eq.) while maintaining the internal temperature below 20°C ± 5°C. After filling is complete, warm container C to 30°C ± 5°C. Then, transfer the contents of container A to container C over 4 hours, maintaining the internal temperature at 30°C ± 5°C. Rinse container A with toluene-2 ​​(0.5 L / kg) and transfer it to container C. Stir the contents of container C at 30°C for an additional 3 hours. Cool the contents of container C to 20°C ± 5°C. Prepare a solution of (S)-1-boc-3-methylpiperidinium (1.2 equiv.) and N,N-diisopropylethylamine-2 (1.2 equiv.) in isopropyl acetate-1 (1.0 L / kg) in container D. Charge the solution from container D into container C, maintaining the batch temperature at 20°C ± 5°C (Note: an exotherm is observed). After the transfer is complete, rinse container D with additional dichloromethane (1.0 L / kg) and transfer it to container C. The contents of Container C were stirred at 20°C for an additional 60 minutes. A solution of sodium bicarbonate [water (15.0 L / kg) + sodium bicarbonate (4.5 equivalents)] was then added to Container C over one hour, maintaining an internal temperature of 20°C ± 5°C throughout the addition. The contents of Container C were stirred for at least 12 hours, at which point the pipazoline (Compound 6) product was isolated by filtration in a stirred filter dryer. The filter cake was washed with water (2) and (3) (5.0 L / kg x 2, stirring for 15 minutes each wash) and isopropyl acetate (2) and (3) (5.0 L / kg x 2, stirring for 15 minutes each wash). The filter cake was dried under nitrogen for 12 hours. [] [Acetone reslurry (if necessary):]

[0147] Pipazoline (Compound 6) and acetone (10.0 L / kg) were charged into vessel E. The suspension was heated to 50°C for 2 hours. Water-4 (10.0 L / kg) was charged into vessel E over 1 hour. After the water addition was complete, the mixture was cooled to 20°C over 1 hour. The contents of vessel E were filtered to isolate the product, and the filter cake was washed with a 1:1 acetone / water mixture (5.0 L / kg). The filter cake was dried under nitrogen for 12 hours. [] [step] [6] [General Note:] All equivalents and volumes refer to the Pipazoline input report [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [or] [kg / kg] [enter] Pipazoline (Compound 6) 531 1.0 equivalent - Boroxine (Compound 6 A) N / A 413.71 0.5 equivalent - Boroxine (Compound 6A) N / A 413.71 0.1 equivalent - 2-Methyltetrahydrofuran 96-47-9 86.13 - 9.0 L / kg 2-Methyltetrahydrofuran 96-47-9 86.13 - 0.5 L / kg Pd(dpePhos)Cl 2 205319-06-8 715.9 0.003 equivalent - Pd(dpePhos)Cl 2 205319-06-8 715.9 0.001 equivalent - Wet 2-methyltetrahydrofuran 96-47-9 86.13 - 4.5 L / kg water 7732-18-5 18.02 - 6.5 L / kg Potassium acetate 127-08-2 98.14 2.0 equivalent - Biaryl Seed 606.7 - 0.002 kg / kg Wet 2-methyltetrahydrofuran 96-47-9 86.13 - 0.02 L / kg heptane 142-82-5 100.20 5.0 L / kg water 7732-18-5 18.02 - 5.0 L / kg Isopropyl alcohol 67-63-0 66.10 - 2.5 L / kg water 7732-18-5 18.02 - 2.5 L / kg Isopropyl alcohol 67-63-0 66.10 - 2.5 L / kg Isopropyl alcohol 67-63-0 66.10 - 2.5 L / kg heptane 142-82-5 100.20 2.5 L / kg Note: All L / kg and kg / kg amounts are relative to the Pipazoline input.

[0148] Reactor A was charged with a solution of pyriprazole (Compound 6, 1.0 equiv), degassed 2-MeTHF (9.0 L / kg), and potassium acetate (2.0 equiv) in degassed water (6.5 L / kg). The resulting mixture was warmed to 75°C ± 5°C and then charged with a slurry of Pd(dpePhos)Cl2 (0.003 equiv) in 2-MeTHF (0.5 L / kg). Within 2 h of the catalyst charge, a solution of freshly prepared boroxine (Compound 6A, 0.5 equiv) in wet degassed 2-MeTHF (4.0 L / kg, KF > 4.0%) was added over a period of >1 hour but <2 hours. After the addition was complete, the reactor was rinsed with an additional portion of wet 2-MeTHF (0.5 L / kg). After the reaction is complete (<0.15 area % pipazoline remains, typically <1 h after the boroxine addition is complete), 0.2 wt % (0.002 kg / kg) biaryl seeds are added as a slurry in 0.02 L / kg wet 2-MeTHF and the resulting seed bed is aged for >60 min. At 75°C ± 5°C, heptane (5.0 L / kg) is added over 2 hours. The batch is then cooled to 20°C ± 5°C over 2 hours and aged for an additional 2 hours. The slurry is then filtered and the filter cake is washed with 1 x 5.0 L / kg water, 1 x 5.0 L / kg 1:1 iPrOH:water, followed by 1 x 5.0 L / kg 1:1 iPrOH:heptane (resuspension wash: resuspend the filter cake with a stirrer and allow to settle before filtering). The filter cake (biaryl, compound 7) was then dried under vacuum with a nitrogen purge. NOTE: If the reaction stops, refill with catalyst and boroxine. [] [step] [7], Charcoal filtration for Pd removal [] General Note: All equivalents and volumes are referenced to the crude biaryl input report [] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [or] [kg / kg] [enter] Initial dissolution Crude Lianfang N / A 606.67 1.0 - dichloromethane 75-09-2 84.93 - 10 L / kg 3M "Zeta Plus R55SP" carbon plate rinse dichloromethane 75-09-2 84.93 - 1.0 L / kg Note: All L / kg and kg / kg values ​​are relative to the crude biaryl input.

[0149] In a clean container A, charge the crude biaryl (1 equivalent) and DCM (10 L / kg). Stir the contents at 22°C ± 5°C for > 60 minutes, observing dissolution. The crude biaryl from container A is passed through a bag filter and a carbon filter at a rate of ≤ 3 L / min / m, and the filtrate is collected in a clean container B. A DCM rinse (1 L / kg) is charged to container A and collected in container B through the carbon filter.

[0150] A sample of the filtrate in container B was taken for IPC Pd analysis. The sample was concentrated to a solid and analyzed by ICP-MS. IPC: Pd ≤ 25 ppm relative to the biaryl. If the Pd content is greater than 25 ppm relative to the biaryl on either the first or second IPC sample, pass the solution through the carbon filter a second time at ≤ 3 L² / min / m², rinsing with 1 L / kg DCM; use the sample filtrate for IPC. b. If the Pd content is still greater than 25 ppm after the third IPC, install and condition a fresh carbon disk. Pass the biaryl filtrate through the new carbon filter and wash with 1 L / kg DCM. Use the sample for IPC.

[0151] Prepare the recovered filtrate for distillation by concentrating to ≤ 4 L / kg DCM and refilling to 5.25 ± 0.25 L / kg DCM before moving to Step 7 Boc-deprotection reaction. [] [step] [7] General Note: All equivalents and volumes are referenced to the crude biaryl input report [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [or] [kg / kg] [enter] [Biaryl] Compound 8 NA 606.67 1.0 - dichloromethane 74-09-2 84.93 - 5.0 L / kg TFA 76-05-1 114.02 15.0 1.9 L / kg potassium carbonate 584-08-7 138.2 18.0 4.1 kg / kg water 7732-18-5 18.02 - 20.0 L / kg 1-Methyl-2-pyrrolidone 872-50-4 99.13 - 1.0 L / kg dichloromethane 74-09-2 84.93 - 1.0 L / kg water 7732-18-5 18.02 - 10.0 L / kg water 7732-18-5 18.02 - 10.0 L / kg Note: All L / kg and kg / kg values ​​are relative to the biaryl input.

[0152] To reactor A was added: tert-butyl (3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperidinium-1-carboxylate (biaryl) (1.0 equiv), dichloromethane (5.0 L / kg), and TFA (15.0 equiv, 1.9 L / kg) was slowly charged to maintain the internal temperature at 20° C. ± 5° C. The reaction was stirred at 20° C. ± 5° C. for 4 h.

[0153] To reactor B, add potassium carbonate (18.0 equivalents), water (20.0 L / kg), and NMP (1.0) to form a homogeneous solution. While stirring at the maximum acceptable rate of the equipment, transfer the reaction mixture in A to the potassium carbonate solution in B over 30 minutes (at a rate of approximately 0.24 L / kg / min). Stir the mixture at 20°C ± 5°C for an additional 12 hours.

[0154] The resulting slurry was filtered and rinsed with water (2 x 10 L / kg). The wet cake was dried for 24 h to afford 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperidin-1-yl]-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (Des-Boc, Compound 8). [] [step] [8] General Note: All equivalents and volumes refer to Des-Boc input reports [] [] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [or] [kg / kg] [enter] reaction Des-Boc compound 8 N / A 506.6 1.0 - TFA 76-05-1 114.02 1.0 1-Methyl-2-pyrrolidone ‎872-50-4 99.1 - 4.2 1-Methyl-2-pyrrolidone 872-50-4 99.1 - 0.8 Acryloyl chloride 814-68-6 90.5 1.3 - Quenching solution Disodium hydrogen phosphate 7558-79-4 142.0 3.0 eq [] water 7732-18-5 18.0 - 15.0 L / kg AMG 510 seeds N / A 560.6 0.005 0.005 kg / kg water 7732-18-5 18.0 - 0.4 L / kg rinse 1-Methyl-2-pyrrolidone 872-50-4 99.1 - 0.5 L / kg washing water 7732-18-5 18.02 - 10.0 L / kg Note: All L / kg and kg / kg values ​​are relative to Des-Boc input

[0155] Under nitrogen, Des-Boc (Compound 8, 1.0 equiv) and NMP (4.2 L / kg) were charged to Container A, and TFA (1.0 equiv) was slowly added to maintain the temperature at <25°C. The mixture was aged at 25°C until complete dissolution was observed (approximately 0.5 h). The solution was then finely filtered through a 0.45 μm filter into Container B and washed with NMP (0.8 L / kg). The filtrate and washings were combined and then cooled to 0°C. Acryloyl chloride (1.3 equiv) was added to the resulting solution while maintaining the temperature at <10°C. The reaction mixture was then aged at 5°C ± 5°C until complete by IPC (approximately 1.5 h). Preparation of disodium phosphate aqueous quenching solution:

[0156] Disodium phosphate (3.0 equivalents) and water (15.0 L / kg) were charged to Vessel C. The mixture was aged at 25°C until complete dissolution was observed. The solution was warmed to 45°C ± 5°C. A seed slurry of AMG 510 (0.005 equivalents) in water (0.4 L / kg) was prepared and added to Vessel C while maintaining the temperature at 45°C ± 5°C.

[0157] The reaction mixture in vessel B was transferred to vessel C (quench solution) while maintaining the temperature at 45°C ± 5°C (approximately 1 hour). Vessel B was washed with a portion of NMP (0.5 L / kg). The product slurry was aged at 45°C ± 5°C for 2 hours, cooled to 20°C over 3 hours, aged at 20°C for a minimum of 12 hours, filtered, and washed with water (2 x 10.0 L / kg). The product was dried under nitrogen and vacuum to yield crude AMG 510 (Compound 9A). [] [step] [9] [General Note:] All equivalent weights and volumes refer to the crude AMG 510 input report [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [or] [kg / kg] [enter] Rough AMG 510 Compound 9A NA 560.60 1.0 - ethanol 64-17-5 - 7.5 L / kg water - 18.02 - 1.9 L / kg AMG 510 seeds[1] - 560.60 0.015 0.015 kg / kg water -- 18.02 - 15.0 L / kg Ethanol (for washing) 64-17-5 - 2.5 V 2.5 L / kg Water (for washing) - - 5.0 V 5.0 L / kg Note: All L / kg and kg / kg values ​​are relative to the crude AMG 510 input. []

[0158] Reactor A was filled with 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1 M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2 S)-2-methyl-4-(prop-2-enyl)piperidin-1-yl]pyrido[2,3-d]pyrimidin-2(1 H)-one ( [ , thick , ] [ , AMG 510 , ]) (1.0 equivalent), ethanol (7.5 L / kg), and water (1.9 L / kg). The mixture was heated to 75°C and finely filtered into a clean reactor B. The solution was cooled to 45°C and inoculated with authentic ground AMG 510 seeds (0.015 ± 0.005 kg / kg); the resulting slurry was aged for 30 minutes. Water (15.0 L / kg) was added over 5 hours while maintaining the internal temperature > 40°C; the mixture was aged for an additional 2 hours.

[0159] The mixture was cooled to 20°C over 3 hours and aged for 8 hours, after which the solid was collected by filtration and washed with a mixture of ethanol (2.5 L / kg) and water (5.0 L / kg). The solid was dried using vacuum and nitrogen to obtain 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1 M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2 S)-2-methyl-4-(prop-2-enyl)piperidin-1-yl]pyrido[2,3- d]pyrimidin-2(1 H)-one ( [ , AMG 510 , ], [ , Compound , ] [ , 9 , ]). [] [Compound] [6A] [Boroxine Synthesis:] [] Lithiation / boration [] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [ / ] [volume] [mol] [quality(] [g] [)] [volume(] [L] [)] 3-Fluoroanisole 456-49-5 126.13 1.0 1.19 150 0.136 n-Butyllithium (2.5 M in hexane) (first base) 109-72-8 64.06 1.5 1.78 N / A 0.712 Diisopropylamine (secondary amine base) 108-18-9 101.19 1.4 1.66 168 0.233 Triethylamine hydrochloride (catalyst) 554-68-7 137.65 0.01 0.012 1.65 N / A Triethyl borate (reagent) 150-46-9 145.99 2.0 2.38 347.5 0.405 Tetrahydrofuran 109-99-9 72.11 12 volume N / A N / A 1.8 Hydrochloric acid (2N) (acid) 7647-01-0 36.46 10 volume N / A N / A 1.5 Methyl tertiary butyl ether 1634-04-4 88.15 12 volume N / A N / A 1.8 heptane 142-82-5 100.20 10.5 volume N / A N / A 1.575

[0160] Reactor A is charged with THF (6 vol), a secondary amine base, diisopropylamine (1.4 eq), and a catalyst, such as triethylamine hydrochloride (0.01 eq). The resulting solution is cooled to -70°C, and the first base, n-BuLi (2.5 M in hexane, 1.5 eq), is slowly added. After the addition is complete, a solution of 3-fluoroanisole (1.0 eq) in THF (6 vol) is slowly added and held at -70°C for 5 min. Simultaneously or sequentially, reagent B (EtO) 3 (2.0 eq) is slowly added and held at -70°C for 10 min. The reaction mixture is quenched with acid, 2N HCl. The quenched reaction mixture is extracted with MTBE (3 x 4 vol). The combined organic phases are concentrated to 1.5-3 vol total. Heptane (7-9 vol) is added dropwise, and the mixture is cooled to 0-10°C and stirred for 3 h. The mixture is filtered and rinsed with heptane (1.5 vol). The solid was dried at < 30 °C under nitrogen to give (2-fluoro-6-methoxyphenyl)boronic acid. [] [Demethylation:] [] [Material] [CAS #] [MW] [(] [g / mol] [)] [equivalent] [L / kg] [or] [kg / kg] [enter] [(2-] [fluorine] [-6-] [Methoxyphenyl] [)] [Boric acid] 78495-63-3 169.95 1.0 - Boron tribromide 10294-33-4 250.52 1.2 1.8 kg / kg dichloromethane 74-09-2 84.93 - 4.0 L / kg dichloromethane 74-09-2 84.93 - 4.0 L / kg water 7732-18-5 18.02 - 3.0 L / kg water 7732-18-5 18.02 - 3.0 L / kg saturated sodium bicarbonate solution As needed dichloromethane 74-09-2 84.93 - 5.0 L / kg concentrated hydrochloric acid As needed water 7732-18-5 18.02 - 3.0 L / kg water 7732-18-5 18.02 - 3.0 L / kg ethanol water water 7732-18-5 18.02 - 3.0 L / kg water 7732-18-5 18.02 - 3.0 L / kg Note: All L / kg and kg / kg amounts are relative to (2-fluoro-6-methoxyphenyl)boronic acid input

[0161] A reactor was charged with dichloromethane (solvent, 4.0 L / kg) and the acid, BBr (1.2 equivalents), and cooled to -20°C. To this solution, a suspension of (2-fluoro-6-methoxyphenyl)boronic acid (1.0 equivalents) in dichloromethane (4.0 L / kg) was added to the BBr / DCM mixture while maintaining the temperature between -15°C and -25°C. The reaction was allowed to proceed for approximately 2 hours while monitoring by HPLC (≤ 1% (2-fluoro-6-methoxyphenyl)boronic acid), after which it was back-quenched into water (3.0 L / kg). The precipitated solid was then isolated by filtration and slurried on the filter with water (3.0 L / kg), followed by stripping. The filtrate was adjusted to pH 4-6 by adding sodium bicarbonate. The bottom organic phase was separated, and the resulting aqueous layer was washed with dichloromethane (solvent, 5.0 volumes) and adjusted to pH = 1 by the addition of concentrated hydrochloric acid. The resulting solid was isolated by filtration, and the filter cake was washed with water (2 x 5.0 L / kg). Purification by reslurry (if necessary)

[0162] The combined crude solids were charged to a reactor and slurried with 5% EtOH / water (5.0 L / kg) for >1 h at 20°C. The purified product was then isolated by filtration and rinsed with water (2 x 3 L / kg) before drying on the filter with nitrogen / vacuum at <30°C to yield 2,2',2''-(1,3,5,2,4,6-trioxatriborane-2,4,6-triyl)tris(3-fluorophenol) ( [Boroxine] [, compound] [6A]). []

[0163] The foregoing is merely illustrative of the present invention and is not intended to limit the invention to the disclosed uses. Variations and modifications that are routine for those skilled in the art are intended to fall within the scope and nature of the invention as defined by the appended claims. All references, patents, applications, and publications cited are incorporated herein by reference in their entirety as if written herein.

[0164] none

[0165] none

Claims

1. A method for preparing a compound having formula (4a), (4a) comprising mixing 1.0 molar equivalent of compound (4) having formula (4) with 3.0 molar equivalent of (+)-2,3-dibenzoyl-D-tartaric acid in the presence of 2-methyltetrahydrofuran to form a compound having formula (4a).

2. The method of claim 1, wherein the compound having formula (4) is mixed with (+)-2,3-dibenzoyl-D-tartaric acid in a solvent, wherein the solvent comprises 2-methyltetrahydrofuran, heptane, or a combination thereof.

3. The method of claim 2, wherein the solvent comprises 2-methyltetrahydrofuran.

4. The method of claim 3, wherein the solvent comprises heptane.

5. A method for preparing a compound having formula (9) or a pharmaceutically acceptable salt thereof, (9) comprising the method of any one of claims 1 to 4. [1] Seeds perform best when ground by a grinding tool or, if no grinding tool is available, when other types of mechanical grinding are used to reduce the particle size. The actual amount of seeds used will be based on seed availability. The target seed quantity is 1.0%-2.0%.

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