Improved Synthesis of KRAS G12C Inhibitor Compounds

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-trioxatriborane-2,4,6-triyl)tris(3-fluorophenol), which can be used in the synthesis of compounds for treating cancers with KRAS G12C mutations, such as compound 9.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 935,502, filed on November 14, 2019, which is incorporated herein by reference in its entirety. Technical field

[0003] This disclosure relates to an improved, efficient, and scalable method for preparing intermediate compounds having a structure such as a compound of Formula 6A, which intermediate compounds can be used in the synthesis of compounds for the treatment of cancers with KRAS G12C mutations. Background art

[0004] KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and cholangiocarcinoma. KRAS mutations are also observed in approximately 25% of NSCLC patients, and some studies have indicated that KRAS mutations are negative prognostic factors in NSCLC patients. Recently, it has been found that mutations in the V - Ki - ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) confer resistance to epidermal growth factor receptor (EGFR) - targeted therapies in colorectal cancer; thus, the mutational status of KRAS can provide important information prior to prescribing TKI therapies. Overall, there is a need for new medical treatments for patients with pancreatic cancer, lung adenocarcinoma, or colorectal cancer, especially those patients who have been diagnosed with such cancers characterized by KRAS mutations and including those who have progressed after chemotherapy. Summary of the invention

[0005] This disclosure relates to the improved preparation of compounds having the following chemical structure:

[0006] Detailed description

[0007] Definitions

[0008] Abbreviations: The following abbreviations may be used herein:

[0009]

[0010]

[0011]

[0012] Unless otherwise indicated, in the context of describing the present invention (especially in the context of the claims), the use of the terms "a", "an", "the", and similar references shall be construed to cover both the singular and the plural. Statements of ranges of values herein are 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. Any and all examples, or exemplary language (e.g., "such as") provided herein are intended to better illustrate the invention and are not a limitation on the scope of the invention. No language in the specification should be construed as indicating any non-claimed element as essential for the practice of the invention.

[0013] As used herein, the term "alkyl" refers to straight-chain and branched-chain C1-C 8 hydrocarbyl groups, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-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 C m-n means an alkyl group having "m" to "n" carbon atoms. The term "alkylene" refers to an alkyl group having substituents. An alkyl group (e.g., methyl) or an alkylene group (e.g., -CH 2 -) group may be substituted by one or more, and typically one to three, groups independently selected from, for example, the following: halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, -NC, amino, -CO 2 H, -CO 2 C 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. The term "haloalkyl" particularly refers to an alkyl group in which at least one (e.g., 1 to 6) or all of the hydrogens of the alkyl group are replaced by halo atoms.

[0014] The terms "alkenyl" and "alkynyl" respectively denote an alkyl group further including a double bond or a triple bond.

[0015] As used herein, the term "halo" refers to fluorine, chlorine, bromine, and iodine. The term "alkoxy" is defined as -OR, where R is an alkyl group.

[0016] As used herein, the term "amino" or "amine" refers interchangeably to an -NR 2 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 NR 3 + . The ammonium moiety is specifically included in the definition of "amino" or "amine". The substituent can be, for example, an alkyl group, an alkoxy group, a cycloalkyl group, a heterocycloalkyl group, an amide group, or a formate group. The R group can be further substituted, for example, by one or more (e.g., one to four) groups selected from the following: halo, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, formate, amine, and amide. The "amide" or "acylamino" group refers interchangeably to a group similar to an amine or amino group but further including C(O), such as -C(O)NR 2 .

[0017] As used herein, the term "aryl" refers to a C 6-14 monocyclic or polycyclic aromatic group, preferably a C 6-10 monocyclic or bicyclic aromatic group, or a C 10-14 polycyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulyl, anthracenyl, phenanthrenyl, pyrenyl, biphenyl, and terphenyl. Aryl also refers to a C 10-14 bicyclic and tricyclic carbocycle, where one ring is aromatic and the other rings are saturated, partially unsaturated, or aromatic, such as dihydronaphthyl, indenyl, indanyl, or tetrahydronaphthyl (tetralinyl). Unless otherwise indicated, the aryl group can be unsubstituted or substituted by one or more, and particularly one to four, groups independently selected from, for example, the following: 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 2 H, -CO 2 C 1 -C 8 alkyl, -OCOC 1 -C 8 alkyl, C 3 -C 10 cycloalkyl, C 3 -C 10 heterocycloalkyl, C 5 -C10 Aryl and C 5 -C 10 heteroaryl.

[0018] As used herein, the term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic carbocyclic ring, wherein the rings of the polycyclic ring can be fused, bridged or spiro. The carbocyclic ring can have 3 to 10 carbocyclic ring atoms. Carbocyclic rings contemplated include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl.

[0019] As used herein, the term "heterocycloalkyl" means a monocyclic or polycyclic (e.g., bicyclic), saturated or partially unsaturated ring system that contains 3 or more (e.g., 3 to 12, 4 to 10, 4 to 8 or 5 to 7) total atoms, wherein 1-5 (e.g., 1, 2, 3, 4 or 5) atoms are independently selected from nitrogen, oxygen and sulfur. Non-limiting examples of heterocycloalkyl include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, dihydropyrrolyl, morpholinyl, thiomorpholinyl, dihydropyridinyl, oxepanyl, dioxepanyl, thiepanyl and diazepanyl.

[0020] Unless otherwise indicated, the cycloalkyl or heterocycloalkyl group can be unsubstituted or substituted with one or more and in particular one to four groups. Some substituents contemplated include halo, C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, -OCF 3 、-NO 2 、-CN、-NC、-OH, alkoxy, amino, -CO 2 H, -CO 2 C 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.

[0021] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic (e.g., bicyclic) system containing one to three aromatic rings and containing one to four (e.g., 1, 2, 3 or 4) heteroatoms selected from nitrogen, oxygen and sulfur in the aromatic rings. In certain embodiments, the heteroaryl has 5 to 20, 5 to 15, 5 to 10 rings, or 5 to 7 atoms. Heteroaryl also refers to C 10-14Bicyclic and tricyclic rings, where one ring is aromatic and the other rings are saturated, partially unsaturated or aromatic. Examples of heteroaryl include, but are not limited to, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl, benzofuryl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzothiophenyl, benzotriazolyl, benzoxazolyl, furanopyridyl, imidazopyridyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuryl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purinyl, pyridinopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidinyl and thiophenopyridyl. Unless otherwise indicated, heteroaryl groups may be unsubstituted or substituted with one or more, and in particular one to four or one or two substituents. Substituents contemplated include halo, C 1-8 alkyl, C 2-8 alkenyl, C 2 - 8 alkynyl, -OCF 3 、-NO 2 、-CN、-NC、-OH, alkoxy, amino, -CO 2 H, -CO 2 C 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.

[0022] As used herein, the term Boc refers to the structure

[0023] Example

[0024] Example 1

[0025] In one embodiment of the present disclosure, the present disclosure includes a compound having Formula 6A:

[0026]

[0027] Example 2

[0028] In another embodiment of the present disclosure, the present disclosure encompasses a composition comprising a compound having Formula 6A:

[0029]

[0030] Example 3

[0031] In another embodiment of the present disclosure, the present disclosure includes a method for preparing a compound having Formula 6A:

[0032]

[0033] The method comprises reacting a mixture comprising a compound having the following structure: and an acid with at least one solvent.

[0034] Example 4

[0035] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3, wherein the acid is BBr 3 .

[0036] Example 5

[0037] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3, wherein the at least one solvent is dichloromethane.

[0038] Example 6

[0039] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3, wherein the at least one solvent is heptane.

[0040] Example 7

[0041] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3, wherein the mixture is cooled to about -20 °C.

[0042] Example 8

[0043] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3, wherein the method for preparing a compound having the following structure: comprises mixing a compound having the following structure: with a reagent, a first base, a secondary amine base, a catalyst, and an acid.

[0044] Example 9

[0045] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 8, wherein the first base is n-butyllithium.

[0046] Example 10

[0047] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 8, wherein the secondary amine base is diisopropylamine.

[0048] Example 11

[0049] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 8, wherein the catalyst is triethylamine hydrochloride.

[0050] Example 12

[0051] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 8, wherein the reagent is triethyl borate.

[0052] Example 13

[0053] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 8, wherein the acid is HCl.

[0054] Example 14

[0055] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3,

[0056] wherein the compound having Formula 6A is used to produce a compound having Formula 7:

[0057]

[0058] Example 15

[0059] In another embodiment of the present disclosure, the present disclosure includes a method for preparing a compound having Formula 7:

[0060]

[0061] The method includes the following steps: reacting a compound having Formula 6A:

[0062]

[0063] with a compound having Formula 6:

[0064]

[0065] in the presence of Pd(dpePhos)Cl 2 and KOAc.

[0066] Example 16

[0067] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 3, wherein the compound having Formula 6A is used to produce a compound having Formula 9:

[0068]

[0069] Example 17

[0070] In another embodiment of the present disclosure, the present disclosure includes the method as described in Example 16, wherein the method further comprises mixing a compound of Formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

[0071] The compounds of the present disclosure

[0072] Provided herein are KRAS inhibitors having the structures discussed in more detail below.

[0073] The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds, wherein one or more atoms of the compounds disclosed herein are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number typically 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 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I. These radiolabeled compounds can be used to assist in determining or measuring the effectiveness of a compound by characterizing, for example, the site or mode of action, or the binding affinity to a pharmacologically important site of action. Certain isotopically labeled compounds of the present disclosure (e.g., those incorporating a radioactive isotope) can be used in drug and / or substrate tissue distribution studies. Given the ease of incorporation and ready means of detection, the radioactive isotopes tritium (i.e., 3 H) and carbon-14 (i.e., 14 C) are particularly useful for this purpose.

[0074] Replacement with a heavier isotope (e.g., deuterium, i.e., 2 H) can provide certain therapeutic advantages (e.g., an extended in vivo half-life or a reduced dosage requirement) resulting from greater metabolic stability, and is thus preferred in some cases.

[0075] With a positron emitting isotope (e.g.,11 C, 18 F, 15 O and 13 N)-substituted 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 similar to those described in the Preparations and Examples described below, using appropriate isotopically labeled reagents in place of the previously employed unlabeled reagents.

[0076] Isotopically labeled compounds as disclosed herein can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the attached examples and protocols, using appropriate isotopically labeled reagents in place of the previously employed unlabeled reagents.

[0077] Certain compounds as disclosed herein can exist as stereoisomers (i.e., isomers that differ only in the spatial arrangement of atoms), including optical isomers and conformational isomers (or conformers). Compounds as disclosed herein include all stereoisomers as pure individual stereoisomeric preparations and enriched preparations of each, as well as racemic mixtures of such stereoisomers and individual diastereomers and enantiomers that can be separated by methods known to those skilled in the art. Additionally, compounds as disclosed herein include all tautomeric forms of these compounds.

[0078] Certain compounds as disclosed herein can exist as atropisomers, which are conformational stereoisomers that arise when rotation about a single bond in a molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule. Compounds as disclosed herein include all atropisomers as pure individual atropisomeric preparations, enriched preparations of each, or non-specific mixtures of each. If the rotational barrier about the single bond is high enough and the interconversion between conformations is slow enough, then separation and isolation of the isomeric species can be tolerated. For example, groups such as but not limited to the following groups may exhibit restricted rotation.

[0079] The term "monohydrate" refers to a salt of a compound 9 having approximately one associated water molecule. Those skilled in the art will understand that the exact number of associated water molecules can vary slightly at any given time with variable temperature, pressure, and other environmental influences. All minor variations in the number of associated water molecules are contemplated within the scope of this disclosure.

[0080] The term "dihydrate" refers to a salt of compound 9 having approximately two associated water molecules. Those skilled in the art will understand that the exact number of associated water molecules can vary slightly at any given time with 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.

[0081] The term "cocrystal" refers to a crystalline material containing two or more compounds at ambient temperature (20 °C to 25 °C, preferably 20 °C), wherein at least two are bound together by weak interactions, wherein at least one of the compounds is a cocrystal former and the other 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.

[0082] The term "amorphous form" or "amorphous" refers to a material that lacks long-range order and thus does not exhibit distinct X-ray diffraction peaks (i.e., Bragg diffraction peaks). The XRPD pattern of an amorphous material is characterized by one or more amorphous halos.

[0083] The term "amorphous halo" is an approximately bell-shaped maximum in the X-ray powder pattern of an amorphous substance.

[0084] 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%.

[0085] The term "patient" refers to an animal, such as a dog, cat, cow, horse, sheep, and human. A particular patient is a mammal. The term patient includes both male and female.

[0086] The terms "treating", "treat", or "treatment" and the like include prophylactic (e.g., preventative) and palliative treatment.

[0087] The term "excipient" means any pharmaceutically acceptable additive, carrier, diluent, adjuvant, or other component other than the active pharmaceutical ingredient (API), which is typically incorporated for formulating and / or administering to a patient.

[0088] Pharmaceutical Compositions, Routes of Administration and Administration

[0089] Also provided herein are pharmaceutical compositions that comprise a compound as disclosed herein and a pharmaceutically acceptable excipient, such as a diluent or a carrier. Compounds and pharmaceutical compositions suitable for use in the present invention include those that can be administered in an effective amount of the compound to achieve its intended purpose. The administration of the compound is described in more detail hereinafter.

[0090] Suitable pharmaceutical formulations can be determined by one of ordinary skill in the art based on the route of administration and the desired dosage. See, e.g., Remington's Pharmaceutical Sciences, 1435-712 (18th ed., Mack Publishing Co, Easton, Pennsylvania, 1990). The formulation can affect the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the administered agent. Depending on the route of administration, suitable dosages can be calculated based on body weight, body surface area, or organ size. One of ordinary skill in the art can further refine the calculations required to determine the appropriate therapeutic dosage in a conventional manner, particularly based on the dosage information and assays disclosed herein and pharmacokinetic data obtainable through animal or human clinical trials without undue experimentation.

[0091] The phrases “pharmaceutically acceptable” or “pharmacologically acceptable” refer to molecular entities and compositions that do not produce adverse reactions, allergic reactions, 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 agents, and absorption delaying agents, etc. The use of such excipients for pharmaceutically active substances is well known in the art. Such conventional media or agents are contemplated for use in therapeutic compositions unless any of them is incompatible with the therapeutic composition. Supplementary active ingredients can also be incorporated into the compositions. In an exemplary embodiment, the formulation can 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 hydrogen 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-glutamic acid, 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, copper sulfate, thiamine hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenite, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0092] The compound can 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.

[0093] Pharmaceutically acceptable base addition salts can be formed with metals or amines (such as alkali metals, 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 are sodium, potassium, magnesium, ammonium, calcium or ferric trivalent, etc. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucosamine and procaine.

[0094] Pharmaceutically acceptable acid addition salts include inorganic acid salts or organic acid salts. Examples of suitable acid salts include hydrochloride, formate, acetate, citrate, salicylate, nitrate, phosphate. 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; salts with organic formic acid, sulfonic acid, sulfonic group acid or phosphoric group acid or N-substituted aminosulfonic acid, 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, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, pamoic acid, nicotinic acid or isonicotinic acid; and salts with amino acids, such as the 20 α-amino acids involved in protein synthesis in nature, such as glutamic acid or aspartic acid, and salts 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 cyclohexylsulfamate), or salts with other acidic organic compounds, such as ascorbic acid.

[0095] Pharmaceutical compositions containing the compounds disclosed herein can be manufactured in a conventional manner, for example by conventional mixing, dissolving, granulating, dragee preparation, grinding, emulsifying, encapsulating, entrapping or lyophilization methods. Suitable formulations depend on the chosen route of administration.

[0096] 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 invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained by adding solid excipients to a compound as disclosed herein, optionally grinding the resulting mixture, and processing the granule mixture, if necessary after adding suitable auxiliaries, to obtain tablet or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. Disintegrating agents can be added if desired. Pharmaceutically acceptable ingredients for various types of formulations are well known and can be, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweetening and flavoring agents, coating materials, preservatives, dyes, thickening agents, adjuvants, antimicrobial agents, antioxidants, and carriers for various types of formulations.

[0097] When orally administering a therapeutically effective amount of a compound disclosed herein, the composition is generally 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).

[0098] When administered in tablet form, the composition can additionally contain functional solids and / or solid carriers, such as gelatin or adjuvants. Tablets, capsules, and powders can contain from about 1% to about 95% of the compound, and preferably from about 15% to about 90% of the compound.

[0099] When administered in liquid or suspension form, functional liquids and / or liquid carriers, such as water, petroleum, or oils of animal or vegetable origin, can be added. The liquid form of the composition can further contain physiological saline solutions, sugar alcohol solutions, dextrose or other sugar solutions, or diols. When administered in liquid or suspension form, the composition can contain from about 0.5% to about 90% by weight of a compound disclosed herein, and preferably from about 1% to about 50% of a compound disclosed herein. In one embodiment contemplated, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition can be supplied as a rapidly dissolving solid formulation for dissolution or suspension immediately prior to administration.

[0100] When administering a therapeutically effective amount of the compounds disclosed herein 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, with due consideration for pH, isotonicity, stability, etc., is within the scope of the skill in the art. In addition to the compounds disclosed herein, preferred compositions for intravenous, transdermal or subcutaneous injection generally contain an isotonic vehicle. Such compositions can be prepared for administration as a solution in water of the free base or a pharmaceutically acceptable salt, appropriately mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof, and in oils. Under ordinary conditions of storage and use, these preparations may optionally contain preservatives to prevent the growth of microorganisms.

[0101] Injectable compositions can 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 cases, the form must be sterile and must be fluid to the extent that easy injectability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi, by, for example, the inclusion of a preservative. The carrier can be a solvent or a dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycols, etc.), suitable mixtures thereof, and vegetable oils. In one embodiment contemplated, the carrier is non-aqueous or substantially non-aqueous. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin; by maintaining the desired particle size of the compounds in the case of dispersions; and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many embodiments, the inclusion of isotonic agents, such as sugars or sodium chloride, will be preferred. Prolonged absorption of injectable compositions can be achieved by the use of absorption delaying agents, such as aluminum monostearate and gelatin, in the composition.

[0102] Sterile injectable solutions are prepared by incorporating the active compound in the required amount into a suitable solvent, optionally containing various other ingredients enumerated above, and then filtering sterilizing. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains a basic dispersion medium and other required ingredients from those enumerated above. In the case 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 other required ingredients from its previously sterile-filtered solution.

[0103] It is also possible to prepare slow-release or sustained-release formulations to achieve controlled release of the active compound upon contact with body fluids in the gastrointestinal tract and to provide a substantially constant and effective level of the active compound in the plasma. For example, the release can be controlled by one or more of dissolution, diffusion, and ion exchange. Additionally, slow-release methods can facilitate absorption through saturable or rate-limiting pathways within the gastrointestinal tract. For example, for this purpose, the compound can be encapsulated in a polymeric matrix of a biodegradable polymer, a water-soluble polymer, or a mixture of both, and optionally a suitable surfactant. In this case, encapsulation can mean incorporation of microparticles in the polymeric matrix. Controlled-release formulations are also obtained by encapsulating the dispersed microparticles or emulsified droplets via known dispersion or emulsion coating techniques.

[0104] For administration by inhalation, the compounds of the present disclosure are conveniently delivered in the form of an aerosol spray from a pressurized package or nebulizer using a suitable propellant. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges, such as gelatin, for inhalers or insufflators can be formulated to contain a powder mixture of the compound and a suitable powder matrix, such as lactose or starch.

[0105] 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 forms (e.g., in ampoules or in multi-dose containers) and are added with preservatives. The compositions can take forms such as suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents.

[0106] Pharmaceutical formulations for parenteral administration include aqueous solutions of the compounds in water-soluble form. Additionally, suspensions of the compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound and permit the preparation of highly concentrated solutions. Alternatively, the compositions of the present invention can be in powder form for constitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0107] 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 previously described formulations, the compounds can 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 suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as a sparingly soluble derivative (e.g., as a sparingly soluble salt).

[0108] In particular, the compounds disclosed herein can be administered orally, buccally or sublingually in the form of tablets containing excipients (such as starch or lactose), or in capsules or ovules, either alone or in admixture with excipients, or in the form of elixirs or suspensions containing flavoring or coloring agents. Such liquid formulations can be prepared with pharmaceutically acceptable additives (such as suspending agents). The compounds can also be injected parenterally, such as intravenously, intramuscularly, subcutaneously or intracoronarily. For parenteral administration, the compounds are optimally used in the form of a sterile aqueous solution, which can contain other substances, such as salts or sugar alcohols (such as mannitol) or glucose, to render the solution isotonic with blood.

[0109] For veterinary use, the compounds disclosed herein are administered as suitably acceptable formulations in accordance with normal veterinary practice. The veterinarian can readily determine the most suitable dosing regimen and route of administration for a particular animal.

[0110] In some embodiments, all of the necessary components for using the compounds disclosed herein, either alone or in combination with another agent or intervention conventionally used to treat such a disease, for treating KRAS-related disorders can be packaged into a kit. Specifically, the present disclosure provides a kit for therapeutic intervention of a disease, the kit comprising a packaged set of medicaments, including the compounds disclosed herein and buffers and other components for preparing a deliverable form of the medicaments; and / or a device for delivering such medicaments; and / or any agent for combination therapy with the compounds disclosed herein; and / or instructions for treating the disease packaged with the medicaments. The instructions can be fixed in any tangible medium, such as printed paper, or a computer-readable magnetic or optical medium, or instructions referring to a remote computer data source, such as a World Wide Web page accessible via the Internet.

[0111] "Therapeutically effective amount" means an amount effective to treat or prevent the development of an existing condition or to alleviate an existing condition in a subject being treated. In particular, based on the detailed disclosures provided herein, determination of an effective amount is well within the capabilities of those of ordinary skill in the art. Generally, a "therapeutically effective dose" refers to the amount of a compound that results in the achievement of the desired effect. For example, in a preferred embodiment, a therapeutically effective amount of the compounds disclosed herein will reduce 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.

[0112] The amount of the compound administered can depend on the subject being treated, the subject's age, health condition, gender and weight, the type of concurrent treatment (if any), the severity of the condition, the nature of the desired effect, the mode and frequency of treatment, and the judgment of the prescribing physician. The dosing frequency can also depend on the pharmacodynamic effect on arterial oxygen partial pressure. However, the most preferred dose can be adjusted according to the individual subject, as will be understood by those of ordinary skill in the art and can be determined without undue experimentation. This generally includes adjusting the standard dose (e.g., reducing the dose if the patient has a low body weight).

[0113] Although individual requirements vary, the determination of the optimal range of the effective amount of the 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, typical doses of the compounds of the present disclosure can be from 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, it can be from 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 dose of the compound can be from about 0.1 mg / kg / day to about 50 mg / kg / day, from about 0.05 mg / kg / day to about 10 mg / kg / day, from about 0.05 mg / kg / day to about 5 mg / kg / day, from about 0.05 mg / kg / day to about 3 mg / kg / day, from about 0.07 mg / kg / day to about 3 mg / kg / day, from about 0.09 mg / kg / day to about 3 mg / kg / day, from about 0.05 mg / kg / day to about 0.1 mg / kg / day, from about 0.1 mg / kg / day to about 1 mg / kg / day, from about 1 mg / kg / day to about 10 mg / kg / day, from about 1 mg / kg / day to about 5 mg / kg / day, from about 1 mg / kg / day to about 3 mg / kg / day, from about 3 mg / day to about 500 mg / day, from about 5 mg / day to about 250 mg / day, from about 10 mg / day to about 100 mg / day, from about 3 mg / day to about 10 mg / day or from about 100 mg / day to about 250 mg / day. Such doses can be administered as a single dose or can be divided into multiple doses.

[0114] Methods of using KRAS G12C inhibitors

[0115] The present disclosure provides methods for inhibiting RAS-mediated cell signaling, the methods comprising contacting a cell with an effective amount of one or more of the compounds disclosed herein. Inhibition of RAS-mediated signal transduction can be evaluated and confirmed in a variety of ways known in the art. Non-limiting examples include showing: (a) a decrease in the GTPase activity of RAS; (b) a decrease in GTP-binding affinity or an increase in GDP-binding affinity; (c) an increase in the K dissociation of GTP or a decrease in the K dissociation of GDP; (d) a decrease in the level of signal transduction molecules downstream in the RAS pathway, such as a decrease in the levels of pMEK, pERK or pAKT; and / or (e) a decrease in the binding of the RAS complex to downstream signal transduction molecules, including but not limited to Raf. One or more of the above items can be determined using kits and commercially available assays.

[0116] The present disclosure also provides methods of treating disease conditions using the compounds or pharmaceutical compositions of the present disclosure, the disease conditions including but not limited to conditions (e.g., cancer) affected by G12C KRAS, HRAS or NRAS mutations.

[0117] In some embodiments, methods of treating cancer are provided, the methods comprising administering to a subject in need thereof an effective amount of any one of the foregoing 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 and cholangiocarcinoma.

[0118] In some embodiments, the present disclosure provides methods of treating a disorder in a subject in need thereof, wherein the methods comprise determining whether the subject has a KRAS, HRAS or NRAS G12C mutation, and if it is determined that the subject has a KRAS, HRAS or NRAS G12C mutation, then administering to the subject a therapeutically effective dose of at least one compound as disclosed herein or a pharmaceutically acceptable salt thereof.

[0119] The disclosed compounds inhibit anchorage-independent cell growth and thus have the potential to inhibit tumor metastasis. Accordingly, another embodiment of the present disclosure provides methods of inhibiting tumor metastasis, the methods comprising administering an effective amount of a compound disclosed herein.

[0120] 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). Accordingly, certain embodiments relate to administering the disclosed compounds (e.g., in the form of a pharmaceutical composition) to a patient in need of treatment for a hematological malignancy. Such malignancies include, but are not limited to, leukemia and lymphoma. For example, the presently disclosed compounds can be used to treat diseases such as, for example: acute lymphoblastic 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, these compounds can be used to treat lymphoma, such as Hodgkin lymphoma or non-Hodgkin lymphoma of all subtypes. In various embodiments, these compounds can be used to treat plasma cell malignancies, such as multiple myeloma, mantle cell lymphoma, and Waldenström macroglobulinemia.

[0121] 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 sequences of wild-type human KRAS, HRAS, or NRAS are known in the art (e.g., accession number NP203524).

[0122] Methods for detecting mutations in the KRAS, HRAS, or NRAS nucleotide sequence are known to those of skill in the art. These 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, a sample is evaluated for a 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, direct sequencing methods using specific regions (e.g., exon 2 and / or exon 3) of the KRAS, HRAS, or NRAS gene are used to identify a G12C KRAS, HRAS, or NRAS mutation. This technique will identify all possible mutations in the sequenced region.

[0123] Methods for detecting mutations in KRAS, HRAS, or NRAS proteins are known to those of skill in the art. These methods include, but are not limited to, detecting KRAS, HRAS, or NRAS mutants using binding agents specific for the mutant proteins (e.g., antibodies), protein electrophoresis and Western blotting, and direct peptide sequencing.

[0124] Methods for determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can use a variety of samples. In some embodiments, the sample is taken 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.

[0125] This disclosure also relates to a method of treating a hyperproliferative disorder in a mammal, the method 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, which cancer is, for example, acute myeloid leukemia, adolescent cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical teratoid tumor, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, atypical teratoid tumor, embryonal tumor, embryonal cell carcinoma, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, ductal carcinoma in situ (DCIS) of the extrahepatic ducts, embryonal tumor, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal glioma, Ewing sarcoma, extracranial embryonal cell carcinoma, extragonadal embryonal cell carcinoma, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), embryonal cell carcinoma, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell carcinoma, pancreatic neuroendocrine tumor, 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 carcinoma, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell tumor, mycosis fungoides, myelodysplastic syndrome, myelodysplasia / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma of bone and osteosarcoma, nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid carcinoma, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach / gastric cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, rare childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer or virus-induced cancer. In some embodiments, the method relates to treating a non-cancerous hyperproliferative disorder, such as a benign proliferation of the skin (e.g., psoriasis), restenosis or of the prostate (e.g., benign prostatic hyperplasia (BPH)).

[0126] In some embodiments, the treatment methods relate to treating lung cancer, and these methods include administering to a subject in need an effective amount of any of the above-described compounds (or a pharmaceutical composition comprising the compound). 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, adenoma, carcinoid tumor, and undifferentiated carcinoma.

[0127] The present disclosure further provides methods for modulating the activity of G12C mutant KRAS, HRAS, or NRAS proteins, which are carried out by contacting the protein with an effective amount of a compound of the present disclosure. The modulation can be inhibition or activation of protein activity. In some embodiments, the present disclosure provides methods for inhibiting protein activity, which are carried out by contacting the 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 G12C mutant KRAS, HRAS, or NRAS proteins, which are carried out by contacting cells, tissues, or organs 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), which are carried out 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 percentage of inhibition exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0128] In some embodiments, the present disclosure provides methods for inhibiting the activity of KRAS, HRAS, or NRAS G12C in cells by contacting the cells with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the cells. In some embodiments, the present disclosure provides methods for inhibiting the activity of KRAS, HRAS, or NRAS G12C in tissues by contacting the tissues with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the tissues. In some embodiments, the present disclosure provides methods for inhibiting the activity of KRAS, HRAS, or NRAS G12C in organisms by contacting the organisms with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the organisms. In some embodiments, the present disclosure provides methods for inhibiting the activity of KRAS, HRAS, or NRAS G12C in animals by contacting the animals with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the animals. In some embodiments, the present disclosure provides methods for inhibiting the activity of KRAS, HRAS, or NRAS G12C in mammals by contacting the mammals with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the mammals. In some embodiments, the present disclosure provides methods for inhibiting the activity of KRAS, HRAS, or NRAS G12C in humans by contacting the humans with an amount of a compound of the present disclosure sufficient to inhibit the activity of KRAS, HRAS, or NRAS G12C in the humans. The present disclosure provides methods for treating a subject in need thereof for a disease mediated by the activity of KRAS, HRAS, or NRAS G12C.

[0129] Combination therapy

[0130] The present disclosure also provides methods for combination therapy, wherein an agent known to modulate other pathways or other components of the same pathway, or even an overlapping set of target enzymes, is combined with a compound of the present disclosure or a pharmaceutically acceptable salt thereof. In one aspect, such therapy includes, but is not limited to, combinations of one or more compounds of the present disclosure with chemotherapeutic agents, therapeutic antibodies, and radiation therapy to provide a synergistic or additive therapeutic effect.

[0131] 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, biologic response modifiers, antihormonal agents, angiogenesis inhibitors, and antiandrogens. Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules such as (imatinib mesylate), (carfilzomib), (bortezomib), Casodex (bicalutamide), (gefitinib), Venclexta TM (venetoclax), and Adriamycin TM (doxorubicin), and various chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan TM ); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphaoramide, and trimethylolomelamine; nitrogen mustards such as chlorambucil, chlornaphazine, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, Casodex TM, chromomycin, actinomycin D, daunomycin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, isorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptozotocin, streptomycin, tubercidin, bestatin, zinostatin, zorubicin; 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, thioguanine, tioguanine; pyrimidine analogs such as azacitidine, aza-cytidine, 6-azauridine, carmofur, cytarabine, didoxuridine, doxifluridine, enocitabine, floxuridine, androgens such as calusterone, dromostanolone propionate, thiotrostan, methyltrienolone, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as folinic acid; acetylcysteine; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; colchicine amide; diaziquone; eflornithine; elisidepsin; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazine; procarbazine; PSK; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2”-trichloroethylamine; urethan; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; cytarabine (“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.

[0132] Suitable chemotherapeutic cytomodulators also include antihormonal agents for modulating or inhibiting the action of hormones on tumors, such as antiestrogens, including for example tamoxifen (NolvadexTM )、raloxifene, aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, and toremifene (Fareston); and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platin; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; Navelbine; Novantrone; teniposide; daunomycin; aminopterin; Xeloda; ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO).

[0133] If desired, the compounds or pharmaceutical compositions of the present disclosure can be used in combination with commonly prescribed anti-cancer drugs such as ABVD, Avicine, Abagovomab, Acridine carboxamide, Adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, Alpharadin, Alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, Amonafide, Anthracenedione, Anti-CD22 immunotoxin, Antineoplastic agent, Antitumorigenic herbs, Apaziquone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW 2992, Biricodar, Brostallicin, Bryostatin, Buthionine sulfoximinesulfoximine), CBV (chemotherapy), Calyculin, cell cycle non-specific anti-tumor agent, dichloroacetic acid, Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin, Everolimus, Exatecan, Exisulind, Ferruginol, Forodesine, Fosfestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod, Indolocarbazole, Irofulven, Laniquidar, Larotaxel, Lenalidomide, Lucanthone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Ortataxel, PAC-1, Pawpaw, Pixantrone, proteasome inhibitor, Rebeccamycin, Resiquimod, Rubitecan, SN-38, Salinosporamide A, Sapacitabine, Stanford V, Swainsonine, Talaporfin, Tariquidar, Tegafur-uracil, Temodar, Tesetaxel, Triplatin tetranitrate, tris(2-chloroethyl)amine, Troxacitabine, Uramustine, Vadimezan, Vinflunine, ZD6126 or Zosuquidar.

[0134] The present disclosure further relates to methods of inhibiting abnormal cell growth or treating hyperproliferative disorders in mammals using a combination of a compound or pharmaceutical composition provided herein and radiotherapy. Techniques for administering radiotherapy are known in the art and these techniques can be used in the combination therapies described herein. The administration of the compounds of the present disclosure in such combination therapies can be determined as described herein.

[0135] Radiotherapy can be administered by one or a combination of several methods, including but not limited to external beam therapy, brachytherapy, implant radiation, stereotactic radiosurgery, total body radiotherapy, radiotherapy and permanent or temporary interstitial brachytherapy. As used herein, the term "brachytherapy" refers to radiotherapy delivered by spatially confined radioactive material that is inserted into the body at or near the site of the tumor or other hyperplastic tissue affected. The term is intended to include, without limitation, exposure to radioactive isotopes (e.g., radioactive isotopes of At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32 and Lu). Suitable radiation sources for use as cell conditioners in 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 rays. 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 a solid radionuclide (e.g., Au-198, Y-90). In addition, one or more radionuclides can be encapsulated in a gel or radioactive microspheres.

[0136] The compounds or pharmaceutical compositions of the present disclosure can be used in combination with an amount of one or more substances selected from: anti-angiogenesis agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors or autophagy inhibitors.

[0137] Anti-angiogenic agents can be used in combination with the compounds of the present disclosure and the pharmaceutical compositions described herein. These anti-angiogenic agents are, for example, MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors. Anti-angiogenic agents include, for example, 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 the following patents: WO 96 / 33172, WO 96 / 27583, European Patent Publication EP 0818442, European Patent Publication EP1004578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, WO 98 / 34915, WO 98 / 33768, WO 98 / 30566, European Patent Publication 606046, European Patent Publication 931 788, WO 90 / 05719, WO 99 / 52910, WO99 / 52889, WO 99 / 29667, WO 1999007675, European Patent Publication EP 1786785, European Patent Publication No. EP 1181017, US Publication US 20090012085, US Publication US 5863 949, US Publication US 5861510, and European Patent Publication EP 0780386, all of which are hereby incorporated by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors are those having minimal or no activity in inhibiting MMP-1. More preferably, those that selectively inhibit MMP-2 and / or AMP-9 relative to other matrix metalloproteinases (i.e., MAP-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.

[0138] The compounds of the present invention can also be used in combination therapies with other anti-tumor agents, such as acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, hexamethylmelamine, amifostine, aminolevulinic acid, amrubicin, aza- n3, interferon alfacon-1, interferon α, natural interferon β, interferon β-1a, interferon β-1b, interferon γ, natural interferon γ-1a, interferon γ-1b, interleukin-1β, iobenguane, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte α interferon, leuprorelin, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, milgramostim, mismatched double-stranded RNA, mitoguazone, dibromodulcitol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis-stimulating protein, NSC 631570 octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronic acid, 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 (Re 186) etidronate, RII retinamide, rituximab, romurtide, samarium (153Sm) lexidronam, sargramostim, sizofiran, sobuzoxane, sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachloro decoxide, thalidomide, thymalfasin, thyrotropin α, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, troxipide, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor α, natural bestatin, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, VIRULIZIN, zinostatinstimalamer) or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotides, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), LYM-1-iodine 131 MAb (Techniclone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab, motexafin gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), rupirtec, satraplatin, sodium phenylacetate, sparfosic acid, SRL 172 (SR Pharma), SU5416 (Sugen, Inc. (now Pfizer, Inc.), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyl etiopurpurin, tirapazamine, cancer vaccines (Bemira), melanoma vaccines (New York University), melanoma vaccines (Sloan Kettering Institute), melanoma tumor lysate vaccines (New York Medical College), viral melanoma cell lysate vaccines (Royal Newcastle Hospital), or valspodar.

[0139] The compounds of the present disclosure can be further used in combination with VEGFR inhibitors. Other compounds described in the following patents and patent applications can be used in combination therapies: 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, WO04 / 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.

[0140] In some embodiments, the combination comprises a combination of the composition of the present disclosure with at least one anti - angiogenic agent. Agents include, but are not limited to, chemical compositions prepared synthetically in vitro, antibodies, antigen - binding regions, radionuclides, and combinations and conjugates thereof. The agent can be an agonist, antagonist, allosteric modulator, toxin, or more generally, can be used to inhibit or stimulate its target (e.g., receptor or enzyme activation or inhibition), and thereby promote cell death or prevent cell growth.

[0141] Exemplary anti - angiogenic agents include ERBITUX TM(IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to the kinase domain receptor), anti-VEGF agents (e.g., antibodies or antigen-binding regions that specifically bind VEGF, or soluble VEGF receptors or their ligand-binding regions) (e.g., AVASTIN TM or VEGF-TRAP TM ), 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 TM (gefitinib), TARCEVA TM (erlotinib), anti-Ang1 agents and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind thereto or to its receptor (e.g., Tie2 / Tek)), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto). The pharmaceutical compositions of the present disclosure may also include one or more agents that specifically bind to growth factors and inhibit the activity of growth factors (e.g., antibodies, antigen-binding regions, or soluble receptors), 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".

[0142] Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., US Publication No. 2003 / 0162712; US Patent No. 6,413,932), anti-TWEAK agents (e.g., antibodies or antigen-binding regions that specifically bind, or soluble TWEAK receptor antagonists; see Wiley, US Patent No. 6,727,225), ADAM disintegrin domains for antagonizing the binding of integrin to its ligand (Fanslow et al., US Publication No. 2002 / 0042368), antibodies or antigen-binding regions that specifically bind anti-eph receptors and / or ephrins (US 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), and antibodies or antigen-binding regions that specifically bind the PDGF-BB ligand and PDGFR kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto).

[0143] Other anti-angiogenesis / anti-tumor agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, US5712291); ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783); vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA, now known as CASI Pharamaceutical); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); α-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands); DAC: anti-angiogenesis 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); Metastatin (Enteral, USA); Angiogenesis inhibitor (Tripep, Sweden); Maspin (Sosei, Japan); 2-Methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); Flumetralin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); Platelet factor 4 (RepliGen, USA, EP407122); Vascular endothelial growth factor antagonist (Borean, Denmark); Bevacizumab (pINN) (Genentech, USA); Angiogenesis inhibitor (SUGEN, USA); XL784 (Exelixis, USA); XL 647 (Exelixis, USA); MAb, α5β3 integrin, second generation (Applied Molecular Evolution, USA and MedImmune, USA); Gene therapy, retinopathy (OxfordBioMedica, UK); Enzastaurin hydrochloride (USAN) (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); Angiogenesis inhibitor (Alchemia, Australia); VEGF antagonist (Regeneron, USA); rBPI 21 and BPI-derived anti-angiogenic agent (XOMA, USA); PI 88 (Progen, Australia); Cilengitide (pINN) (Merck KGaA, Germany;Technical University of Munich, 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); ZD6126 (Angiogene 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 (Entremed, USA); Pegaptanib (pINN) (Gilead Sciences, USA); Zingiberone (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); Metastasin (Entremed, USA, now known as CASI Pharmaceuticals); Troponin I (Harvard University, USA); SU 6668 (Sugen, USA, now known as Pfizer); OXI 4503 (OXiGENE, USA);Linguanide (Dimensional Pharmaceuticals, USA); Motuporamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); Atimolol (pINN) (GlaxoSmithKline, UK); E7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); Angiogenesis vaccine (EntreMed, USA, now known as CASI Pharmaceuticals); Urokinase plasminogen activator inhibitor (Dendreon, USA); Oglufanide (pINN) (Melmotte, USA); HIF-1α inhibitor (Xenova, UK); CEP5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); Angiostatin (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); Intravitreal 2-methoxyestradiol drug delivery system (EntreMed, USA); Anginex (Maastricht University, Netherlands and Minnesota University, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA);Tumor necrosis factor-α inhibitor (National Institute on Aging, USA); SU 11248 (Pfizer Inc. and Sugen Inc., USA); ABT 518 (Abbott Laboratories, USA); YH16 (Yantai Rongchang, China); S-3APG (Children's Hospital Boston and EntreMed Inc., USA); MAb, KDR (ImClone Systems, USA); MAb, α5β1 (Protein Design, USA); KDR kinase inhibitor (Celltech Group plc and Johnson & Johnson, USA); GFB 116 (University of South Florida and Yale University, USA); CS 706 (Sankyo Co., Ltd., Japan); Combretastatin A4 prodrug (Arizona State University, USA); Chondroitinase AC (IBEX Technologies Inc., Canada); BAY RES 2690 (Bayer AG, Germany); AGM 1470 (Harvard University, USA, Takeda Pharmaceutical Company Limited, Japan and TAP Pharmaceuticals Products Inc., USA); AG 13925 (Agouron Pharmaceuticals Inc., USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD732 (Chong Kun Dang, South Korea); MAb, vascular endothelial growth factor (Nova Laboratories Limited, UK); Irsogladine (INN) (Nippon Shinyaku Co., Ltd., Japan); RG 13577 (Aventis Pharma S.A., France); WX 360 (Wilex AG, Germany); Squalamine (pINN) (Genaera Corporation, USA); RPI 4610 (Sirna Therapeutics, Inc., USA); Cancer therapy (Marinova Pty Ltd., Australia); Heparanase inhibitor (InSight Biotechnology Ltd., 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 (Shire, USA); VGA 1102 (Taisho, Japan); VEGF receptor modulator (Pharmacopeia, USA); VE-cadherin-2 antagonist (ImClone Systems, USA); Angiostatin (National Institutes of Health, USA); Flk-1 vaccine (ImClone Systems, USA); TZ 93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); Truncated soluble FLT 1 (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).;

[0144] Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil TM )), bafilomycin A1, 5-amino-4-imidazolecarboxamide ribonucleoside (AICAR), okadaic acid, autophagy-inhibiting marine toxins that inhibit protein phosphatase type 2A or type 1, cAMP analogs, and drugs that elevate cAMP levels such as adenosine, LY204002, N6-mercaptopurine ribonucleoside, and vinblastine. Additionally, antisense or siRNA that inhibits protein expression can be used, and these proteins include, but are not limited to, ATG5 (which is involved in autophagy).

[0145] Other pharmaceutically active compounds / agents that can be used to treat cancer and can be used in combination with one or more of the compounds of the present disclosure include: erythropoietin alpha; darbepoetin alfa; panitumumab; pegfilgrastim; palifermin; filgrastim; denosumab; ansermin; AMG 102; AMG 176; AMG 386; AMG 479; AMG 655; AMG 745; AMG 951; and AMG 706, or a pharmaceutically acceptable salt thereof.

[0146] 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, epidipodophyllotoxin (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 metabolizes L-asparagine systemically and deprives cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative / antimitotic alkylating agents (e.g., nitrogen mustards such as mechlorethamine, e.g., dichloromethyldiethylamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimines and methylmelamines (e.g., hexaamethylmelaamine and thiotepa), CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs and streptozocin), triazenes-dacarbazinine (DTIC), antiproliferative / antimitotic antimetabolites such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (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, apicidan, hydroxamic 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), multi-kinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g.,estrogens) and hormonal agonists such as luteinizing hormone releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide, and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK 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., 17AAG and KOS 953), PI3K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTI (e.g., Zarnestra, TM ), 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 can include mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib or any analogs or derivative variants of the foregoing.

[0147] The compounds of the present disclosure can also be used in combination with radiotherapy, hormone therapy, surgery, and immunotherapy, which are well known to those skilled in the art.

[0148] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with a steroid. Suitable steroids can include, but are not limited to, 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoxymethasone, dexamethasone, diflorasone, diflucortolone, difuprednate, glycyrrhetinic acid, flucortolone, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinolone acetonide acetate, fluocortin butyl, flucortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, hydrocortisone, loteprednol etabonate, mapracorat, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednisolone valerate, prednylidene, rimocidin, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and salts and / or derivatives thereof. In a particular embodiment, the compounds of the present disclosure can also be used in combination with other pharmaceutically active agents for treating nausea.Examples of agents useful for treating nausea include: dronabinol; granisetron; metoclopramide; ondansetron; and prochlorperazine; or pharmaceutically acceptable salts thereof.

[0149] The compounds of the present disclosure 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 compounds are PD-1 and PD-L1 antagonists. The compounds or pharmaceutical compositions of the present disclosure may also be used in combination with an amount of one or more substances selected from: 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.

[0150] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful EGFR antibody inhibitors include cetuximab (Erbitux), panitumumab (Vectibix), 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 J G et al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004; 304(5676):1497-500.

[0151] Non-limiting examples of small molecule EGFR inhibitors include any EGFR inhibitor described in the following patent publications, as well as all pharmaceutically acceptable salts and solvates of said EGFR inhibitors: European Patent Application EP520722, published December 30, 1992; European Patent Application EP 566226, published October 20, 1993; PCT International Publication WO 96 / 33980, published October 31, 1996; US 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 August 21, 1997; PCT International Publication WO 97 / 38994, published October 23, 1997; PCT International Publication WO 97 / 49688, published December 31, 1997; European Patent Application EP 837063, published April 22, 1998; PCT International Publication WO 98 / 02434, published January 22, 1998; PCT International Publication WO 97 / 38983, published October 23, 1997; PCT International Publication WO 95 / 19774, published July 27, 1995; PCT International Publication WO 95 / 19970, published 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 January 23, 1997; PCT International Publication WO 97 / 27199, published July 31, 1997; PCT International Publication WO 98 / 07726, published February 26, 1998; PCT International Publication WO 97 / 34895, published 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 WO98 / 17662, published on April 30, 1998; U.S. Patent No. 5,789,427, authorized on August 4, 1998; U.S. Patent No. 5,650,415, authorized on July 22, 1997; U.S. Patent No. 5,656,643, authorized on August 12, 1997; PCT International Publication WO 99 / 35146, published on July 15, 1999; PCT International Publication WO 99 / 35132, published on July 15, 1999; PCT International Publication WO 99 / 07701, published on February 18, 1999; and PCT International Publication WO 92 / 20642, published on November 26, 1992. Other non-limiting examples of small molecule EGFR inhibitors include any EGFR inhibitor described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12):1599-1625.

[0152] Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block the activation of EGFR by its natural ligands. Non-limiting examples of antibody-based EGFR inhibitors include those described in the following literature: Modjtahedi, H. et al., 1993, Br. J. Cancer 67:247-253; Teramoto, T. et al., 1996, Cancer 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S.M. et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang, X. et al., 1999, Cancer Res. 59:1236-1243. Thus, an EGFR inhibitor can be monoclonal antibody Mab E7.6.3 (Yang, 1999, ibid.), or Mab C225 (ATCC accession number HB-8508), or an antibody or antibody fragment having its binding specificity.

[0153] The KRAS of the present disclosure G12C inhibitors can be used in combination with MEK inhibitors. Specific MEK inhibitors that can be used in the combinations of the present disclosure include PD-325901, trametinib, pimasertib, MEK162 [also known as binimetinib], TAK-733, GDC-0973, and AZD8330. Specific MEK inhibitors that can be used in combination with the KRAS G12C inhibitors are trametinib (trade name: 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 combinations 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.

[0154] PI3K inhibitors include, but are not limited to, wortmannin, 17-hydroxy wortmannin analogs described in WO 06 / 044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (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]propanenitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT Publication No. WO 06 / 122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in PCT Publication No. WO 2008 / 070740), LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, available from Axon Medchem), PI 103 hydrochloride (3-[4-(4-morpholinopyridino-[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 Axon Medchem), PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide, available from Axon Medchem), GDC-0941 dimethanesulfonate (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine, available from Axon Medchem), AS-252424 (5-[1-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-methyl-(Z)-methylene]-thiazolidine-2,4-dione, available from Axon Medchem), and TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one, available from Axon Medchem), 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.

[0155] AKT inhibitors include, but are not limited to, Akt-1-1 (which inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (which inhibits Akt1 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., WO05011700); indole-3-carbinol and its derivatives (e.g., U.S. Patent No. 6,656,963; Sarkar and Li (2004) J Nutr., 134 (12 Suppl.), 3493S-3498S); perifosine (e.g., which 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 troxacitabine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al. (2004) Cancer Res., 64, 4394-9).

[0156] TOR inhibitors include, but are not limited to, AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, 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; rapalogs, such as those disclosed in WO 98 / 02441 and WO01 / 14387, such as AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methyl propionate]-rapamycin (also known as CC1779), 40-epi-(tetrazolyl)-rapamycin (also known as ABT578), 32-deoxyrapamycin, 16-pentyloxy-32(S)-dihydrorapamycin, and other derivatives disclosed in WO 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, WO94 / 02136, WO 95 / 16691, WO 96 / 41807, WO 96 / 41807, and U.S. Patent No. 5,256,790; phosphorous-containing rapamycin derivatives (e.g., WO 05016252); 4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional Application No. 60 / 528,340).

[0157] MCl-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. 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 to not only conventional chemotherapy but also targeted therapies including BCL-2 inhibitors (e.g., ABT-263).

[0158] In this disclosure, KRAS G12CThe inhibitor can also be used in combination with an SHP2 inhibitor. SHP2 inhibitors that can be used in the combinations of the present invention include, but are not limited to, SHP099, and RMC-4550 or RMC-4630 (from Revolutions Medicines, Redwood City, CA).

[0159] Proteasome inhibitors include, but are not limited to (carfilzomib), (bortezomib), and oprozomib.

[0160] Immunotherapies include, but are not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents.

[0161] Monoclonal antibodies include, but are not limited to (daratumumab), (trastuzumab), (bevacizumab), (rituximab), (ranibizumab), and (aflibercept).

[0162] Immunomodulatory drugs (IMiDs) are a class of immunomodulatory drugs (drugs that regulate the immune response) containing an imide group. The class of IMiDs includes thalidomide and its analogs (lenalidomide, pomalidomide, and amiselim).

[0163] Anti-PD-1 inhibitors, including but not limited to antibodies, include, but are not limited to, pembrolizumab AMG404, and nivolumab Exemplary anti-PD-1 antibodies and methods of use thereof are described in the following references: 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 / JP2006 / 309606 (Publication No. WO 2006 / 121168 A1), each of which is hereby expressly incorporated by reference herein. Included: Yervoy TMIpilimumab or tremelimumab (against CTLA-4), galiximab (against B7.1), BMS-936558 (against PD-1), MK-3475 (against PD-1), AMP224 (against B7DC), BMS-936559 (against B7-H1), MPDL3280A (against B7-H1), MEDI-570 (against ICOS), AMG557 (against B7H2), MGA271 (against B7H3), IMP321 (against LAG-3), BMS-663513 (against CD137), PF-05082566 (against CD137), CDX-1127 (against CD27), anti-OX40 (Providence Health Services), huMAbOX40L (against OX40L), atacicept (against TACI), CP-870893 (against CD40), lucatumumab (against CD40), dacetuzumab (against CD40), muromonab-CD3 (against CD3), ipilimumab (against CTLA-4). Immunotherapies also include genetically engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTE).

[0164] 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 No.: WO 2010 / 003118 and 2011 / 090754, or anti-GITR antibodies described, for example, in the following: 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.

[0165] The compounds described herein can be used in combination with the agents or other suitable agents disclosed herein, depending on the condition being treated. Thus, in some embodiments, one or more of the compounds disclosed herein will be co-administered with other agents as described above. When used in combination therapy, the compounds described herein are administered simultaneously or separately from the second agent. Such combination administration can include administering both agents simultaneously in the same dosage form, simultaneously in separate dosage forms, and separately. That is, the compounds described herein and any of the above agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds disclosed herein and any of the above agents can be administered simultaneously, where the two agents are present in separate formulations. In another alternative, any of the above agents can be administered immediately after administering the compounds disclosed herein, or vice versa. In some embodiments of the separate administration regimens, the administration of the compounds disclosed herein and any of the above agents is separated by a few minutes, or a few hours, or a few days.

[0166] Since one aspect of the present disclosure contemplates treating a disease / condition with a combination of pharmaceutically active compounds that can be administered separately, the present disclosure further relates to combining individual pharmaceutical compositions in kit form. The kit comprises two individual pharmaceutical compositions: a compound of the present disclosure and a second pharmaceutical compound. The kit comprises containers for holding the individual compositions, such as separate bottles or separate foil pouches. Other examples of containers include syringes, cartridges, and bags. In some embodiments, the kit comprises instructions for use of the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (e.g., oral and parenteral), at different dosage intervals, or when titration of an individual component of the combination is required by the prescribing healthcare professional.

[0167] All patents and other publications cited herein are incorporated herein by reference.

[0168] 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.

[0169] Related synthetic methods

[0170] 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-propenoyl)-1-piperazinyl)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 invention.

[0171] 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, Nov. 22, 2018), which claims the priority of U.S. Provisional Application No. 62 / 509,629, filed May 22, 2017, and claims the benefit of the priority of that provisional application number, and both of these documents are incorporated herein by reference in their entirety for all purposes. 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one is prepared using the following method, in which the isomers of the final product are separated by chiral chromatography.

[0172]

[0173] Step 1: 2,6-Dichloro-5-fluoronicotinamide (Intermediate S). To a mixture of 2,6-dichloro-5-fluoro-nicotinic acid (4.0 g, 19.1 mmol, AstaTech Inc., Bristol, PA) in dichloromethane (48 mL) was added oxalyl chloride (2 M DCM solution, 11.9 mL, 23.8 mmol), followed by a catalytic amount of DMF (0.05 mL). The reaction was stirred overnight at room temperature 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 added dropwise 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, then filtered. The filtered solid was discarded, and the remaining mother liquor portion was concentrated to half volume and filtered. The filtered solid was washed with heptane and dried overnight in a vacuum oven (45 °C) to afford 2,6-dichloro-5-fluoronicotinamide. 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm 8.23 (d, J = 7.9 Hz, 1H) 8.09 (br s, 1H) 7.93 (br s, 1H). m / z (ESI, +ve ion): 210.9 (M + H) + 。

[0174] Step 2: 2,6-Dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. Oxalyl chloride (2 M DCM solution, 14.4 mL, 28.8 mmol) was added dropwise via syringe to a slurry of ice-cooled 2,6-dichloro-5-fluoronicotinamide (Intermediate S, 5.0 g, 23.9 mmol) in THF (20 mL). The resulting mixture was heated at 75 °C for 1 h, then heating was stopped and the reaction was concentrated to half volume. After cooling to 0 °C, THF (20 mL) was added dropwise via cannula, followed by addition of a solution of 2-isopropyl-4-methylpyridin-3-amine (Intermediate R, 3.59 g, 23.92 mmol) in THF (10 mL). The resulting mixture was stirred at 0 °C for 1 h and 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 afford 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. This material was used in the following step without further purification. m / z (ESI, +ve ion): 385.1 (M + H) + 。

[0175] Step 3: 7-Chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione. Potassium bis(trimethylsilyl)amide (1 M THF solution, 50.2 mL, 50.2 mmol) was slowly added via syringe to a solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (9.2 g, 24.0 mmol) in THF (40 mL) cooled in an ice bath. 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 afford 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione. 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm 12.27 (brs, 1H), 8.48 - 8.55 (m, 2H), 7.29 (d, J = 4.8 Hz, 1H), 2.87 (quin, J = 6.6 Hz, 1H), 1.99 - 2.06 (m, 3H), 1.09 (d, J = 6.6 Hz, 3H), 1.01 (d, J = 6.6 Hz, 3H). 19 F NMR (376 MHz, DMSO-d 6 ) δ: -126.90 (s, 1F). m / z (ESI, +ve ion): 349.1 (M + H) + .

[0176] Step 4: 4,7-Dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. 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 afford 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. The material was used in the following step without further purification. m / z (ESI, +ve ion): 367.1 (M + H) + .

[0177] Step 5: (S)-tert-Butyl 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. To a 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) cooled to ice, DIPEA (7.1 mL, 40.3 mmol) was added, followed by (S)-4-N-Boc-2-methylpiperazine (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 were separated, and the aqueous layer was 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), providing (S)-tert-Butyl 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. m / z (ESI, +ve ion): 531.2 (M+H) + 。

[0178] Step 6: (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-methylpiperazine-1-carboxylate. A mixture of (S)-tert-butyl 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (4.3 g, 8.1 mmol), potassium tris(2-fluoro-6-hydroxyphenyl)borate (Intermediate Q, 2.9 g, 10.5 mmol), potassium acetate (3.2 g, 32.4 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (661 mg, 0.81 mmol) in 1,4-dioxane (80 mL) was degassed with nitrogen for 1 min. Deoxygenated water (14 mL) was added and the resulting mixture was heated at 90 °C for 1 h. The reaction was cooled to room temperature, quenched with semi-saturated 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 afford (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-methylpiperazine-1-carboxylate. 1 H NMR (400 MHz, DMSO-d 6 ) δ ppm 10.19 (br s, 1H), 8.38 (d, J = 5.0 Hz, 1H), 8.26 (dd, J = 12.5, 9.2 Hz, 1H), 7.23 - 7.28 (m, 1H), 7.18 (d, J = 5.0 Hz, 1H), 6.72 (d, J = 8.0 Hz, 1H), 6.68 (t, J = 8.9 Hz, 1H), 4.77 - 4.98 (m, 1H), 4.24 (br t, J = 14.2 Hz, 1H), 3.93 - 4.08 (m, 1H), 3.84 (br d, J = 12.9 Hz, 1H), 3.52 - 3.75 (m, 1H), 3.07 - 3.28 (m, 1H), 2.62 - 2.74 (m, 1H), 1.86 - 1.93 (m, 3H), 1.43 - 1.48 (m, 9H), 1.35 (dd, J = 10.8, 6.8 Hz, 3H), 1.26 - 1.32 (m, 1H), 1.07 (dd, J = 6.6, 1.7 Hz, 3H), 0.93 (dd, J = 6.6, 2.1 Hz, 3H). 1919F NMR (376 MHz, DMSO-d 6 ) δ: -115.65 (s, 1F), -128.62 (s, 1F). m / z (ESI, +ve ion): 607.3 (M+H) + .

[0179] Step 7: 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one. 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-methylpiperazine-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 successively with a solution of 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 and then quenched with semi-saturated aqueous sodium bicarbonate 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 afford 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one. 1 1H NMR (400 MHz, DMSO-d 6) δ ppm 10.20 (s, 1H), 8.39 (d, J = 4.8 Hz, 1H), 8.24 - 8.34 (m, 1H), 7.23 - 7.32 (m, 1H), 7.19 (d, J = 5.0 Hz, 1H), 6.87 (td, J = 16.3, 11.0 Hz, 1H), 6.74 (d, J = 8.6 Hz, 1H), 6.69 (t, J = 8.6 Hz, 1H), 6.21 (br d, J = 16.2 Hz, 1H), 5.74 - 5.80 (m, 1H), 4.91 (br s, 1H), 4.23 - 4.45 (m, 2H), 3.97 - 4.21 (m, 1H), 3.44 - 3.79 (m, 2H), 3.11 - 3.31 (m, 1H), 2.67 - 2.77 (m, 1H), 1.91 (s, 3H), 1.35 (d, J = 6.8 Hz, 3H), 1.08 (d, J = 6.6 Hz, 3H), 0.94 (d, J = 6.8 Hz, 3H). 19 19F NMR (376 MHz, DMSO - d 6 ) δ ppm - 115.64 (s, 1F), - 128.63 (s, 1F). m / z (ESI, +ve ion): 561.2 (M + H) + .

[0180] Another synthesis of Compound 9 and related intermediates is described in the U.S. Provisional Patent Application filed on November 16, 2018, which is incorporated herein by reference in its entirety for all purposes.

[0181]

[0182] Representative synthetic methods

[0183] This disclosure includes the following steps, wherein in the manufacture of AMG 510 (Compound 9), the synthesis and utilization of the boroxane intermediate are novel and inventive steps.

[0184]

[0185] Raw materials

[0186]

[0187]

[0188] Step 1a

[0189]

[0190]

[0191]

[0192] Oxalyl chloride (18.9 kg; 148.9 mol) was added 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) 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 h. 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 min and then the product was collected by filtration. The filtered solid was washed with deionized water (3 X 87 L) and dried to afford 2,6-dichloro-5-fluoronicotinamide (Compound 1).

[0193] Step 1b

[0194]

[0195]

[0196] In reactor A, oxalyl chloride (11.9 kg; 93.8 mol) was added to a solution of 2,6-dichloro-5-fluoronicotinamide (Compound 1) (16.27 kg; 77.8 mol) in dichloromethane (359.5 kg) while maintaining the temperature at ≤25 °C for 75 min. The resulting solution was then heated to 40 °C ± 3 °C and aged for 3 h. Using vacuum, the solution was distilled to remove dichloromethane until the solution was below the stirrer. Then dichloromethane (300 kg) was added and the mixture was cooled to 0 ± 5 °C. 2-Isopropyl-4-methylpyridin-3-amine (aniline Compound 2A) (12.9 kg; 85.9 mol) was added to a clean dry reactor (Reactor B), 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 with additional dichloromethane until the solution was dry by KF analysis (limit ≤0.05%). The solution volume was adjusted to approximately 23 L volume with dichloromethane. Then the dry aniline solution was added to reactor A while maintaining the internal temperature at 0 ± 5 °C throughout the addition. The mixture was then heated to 23 °C and aged for 1 h. The solution was finely filtered into a clean reactor to obtain a DCM solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (Compound 3) and used directly in the next step.

[0197] Step 2

[0198]

[0199]

[0200]

[0201] The solvent of the dichloromethane solution of 2,6-dichloro-5-fluoro-N-{[4-methyl-2-(propan-2-yl)pyridin-3-yl]carbamoyl}pyridine-3-carboxamide (UREA (Compound 3)) (containing 15 kg; 38.9 mol) was changed to 2-MeTHF using vacuum distillation while maintaining the internal temperature at 20 °C - 25 °C. The reactor volume was adjusted to 40 L, and then an additional 2-MeTHF (105.4 kg) was charged. Sodium tert-butoxide (9.4 kg; 97.8 mol) was added while maintaining 5 °C - 10 °C. The contents were warmed to 23 °C and stirred for 3 h. Then the contents were 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 it was further aged for 30 min. Stirring was stopped and the layers were separated. The aqueous layer was removed, and deionized water (81.7 L) was added to the organic layer. A mixture of concentrated HCl (1.5 kg) and water (9 L) was prepared and then slowly added to the reactor until the measured pH value was between 4 - 5. The layers were separated, and the aqueous layer was back-extracted with 2-MeTHF (42.2 kg). The two organic layers were combined and washed with a 10% citric acid solution (75 kg), followed by washing with a mixture of water (81.7 L) and saturated NaCl (19.8 kg). Then the organic layer was washed with saturated sodium bicarbonate (75 kg), and if necessary, repeated to achieve an aqueous solution target pH ≥ 7.0. The organic layer was washed again with brine (54.7 kg), and then dried over magnesium sulfate (5 kg). The mixture was filtered to remove magnesium sulfate, and the filter bed was rinsed with 2-MeTHF (49.2 kg). The combined filtrate and washings were distilled 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 h, and then heptane (27.3 kg) was added over 2 h. The product slurry was aged at 20 - 25 °C for 3 h, then filtered and washed with a mixture of 2-MeTHF (2.8 kg) and heptane (9 kg). The product was dried using nitrogen and vacuum to obtain the solid 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (racemic-dione (Compound 4)).

[0202] Step 3

[0203]

[0204]

[0205] Under a nitrogen atmosphere, into a container, (+)-2,3-dibenzoyl-D-tartaric acid (2.0 equivalents) was added to a stirred suspension of Compound 4 (1.0 equivalent) in 2-methyltetrahydrofuran (7.0 L / kg). 2-MeTHF is chiral but is used as a racemic mixture. The different enantiomers of 2-MeTHF are randomly incorporated into the co-crystal. The resulting suspension was warmed to 75 °C and aged at 75 °C until complete dissolution was observed (≤30 min). The resulting solution was filtered through a fine filter into a second container at 75 °C. n-Heptane (2.0 L / kg) was charged into the fine-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. n-Heptane (3.0 L / kg) was charged into the suspension, and then the suspension was aged at 20 °C under a nitrogen atmosphere for 4 hours. 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 with nitrogen and under vacuum to give the diketone:DBTA:Me-THF complex (Compound 4a).

[0206] Step 4

[0207]

[0208]

[0209] In container A, stir a suspension of disodium 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 container B, stir a suspension of the diketone:DBTA:Me-THF complex (Composition 4a) [46.9 kg (corrected to 25.9 kg, 1.0 equivalent for diketone)] in methyl tert-butyl ether (517.8 L, 11.0 L / kg) for 15 to 30 minutes. Add the resulting solution from container A to container B, then stir the mixture for over 3 hours. Stop stirring and separate the biphasic mixture 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 container B and dry with magnesium sulfate (24.8 kg, 0.529 kg / kg). Stir the resulting suspension from container B for over three hours and then filter it into container C. Charge container B with a methyl tert-butyl ether (46.9 L, 1.0 L / kg) rinse and then filter it into container C. Cool the contents of container C to 10 °C and then distill under vacuum while slowly warming to 35 °C. Continue distillation until 320 - 350 kg (6.8 - 7.5 kg / kg) of methyl tert-butyl ether is collected. After cooling the contents of container C to 20 °C, charge n-heptane (278.7 L, 5.9 L / kg) over 1 hour and then distill under vacuum while slowly warming to 35 °C. Continue distillation until 190 - 200 kg (4.1 - 4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane is collected. After cooling the contents of container C to 20 °C, charge n-heptane (278.7 L, 5.9 L / kg) a second time over 1 hour and then distill under vacuum while slowly warming to 35 °C. Continue distillation until 190 - 200 kg (4.1 - 4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane is collected. After cooling the contents of container C to 20 °C, charge n-heptane (195.9 L, 4.2 L / kg) a third time over 1 hour and then sample for GC analysis of the solvent composition. Continue stirring the suspension in container C for over one hour. Filter the suspension and then wash it from container C with an n-heptane (68.6 L, 1.5 L / kg) rinse. Dry the separated solid at 50 °C and submit a sample for storage. Obtain the 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 (diketone) Compound 5M.

[0210] The first-generation method highlighted above has been successfully scaled up to 200+ kg of racemic-dione starting material (Compound 4). In this method, seeding with the thermodynamically stable racemic-dione crystal form (exhibiting low solubility) resulted in batch failures. Based on our subsequent studies, we found that by adjusting the heptane feed schedule to increase the DBTA equivalent and decrease the seed temperature, the robustness of the method was improved. The improved method was resistant to the presence of the thermodynamically stable racemic-dione crystal form and facilitated the successful resolution of the atropisomers. Subsequent batches incorporated the improved method for large-scale manufacturing.

[0211] Step 5

[0212]

[0213]

[0214] Note: All L / kg amounts are relative to the m-dione input; all equivalent amounts are relative to the m-dione input adjusted by potency.

[0215] Charge the m-dione (Compound 5M, 1.0 equivalent) and toluene-1 (10.0 L / kg) into vessel A. Dry the resulting solution under vacuum at 45 °C by azeotropic distillation until 5.0 L / kg of solvent has been removed. Then cool the contents of vessel A to 20 °C.

[0216] Charge vessel C with toluene-3 (4.5 L / kg), phosphoryl chloride (1.5 equivalents), and N,N-diisopropylethylamine-1 (2.0 equivalents) while maintaining the internal temperature below 20 °C ± 5 °C.

[0217] After completion of filling, warm the vessel C to 30 °C ± 5 °C. Then transfer the contents of vessel A to vessel C over 4 hours while maintaining the internal temperature at 30 °C ± 5 °C. Rinse vessel A with toluene-2 (0.5 L / kg) and transfer it to vessel C. Stir the contents of vessel C at 30 °C for an additional 3 hours. Cool the contents of vessel C to 20 °C ± 5 °C. Prepare a solution of (s)-1-boc-3-methylpiperazine (1.2 equivalents), N,N-diisopropylethylamine-2 (1.2 equivalents) in isopropyl acetate-1 (1.0 L / kg) in vessel D. Charge the solution from vessel D into vessel C while maintaining the batch temperature at 20 °C ± 5 °C (Note: Exotherm observed). After the transfer is complete, rinse vessel D with additional dichloromethane (1.0 L / kg) and transfer it to vessel C. Stir the contents of vessel C at 20 °C for an additional 60 minutes. Then charge a solution of sodium bicarbonate [water-1 (15.0 L / kg + sodium bicarbonate (4.5 equivalents)] into vessel C over one hour while maintaining the internal temperature at 20 °C ± 5 °C throughout the addition. Stir the contents of vessel C for at least 12 hours, at which point separate the pipazoline (Compound 6) product by filtration in a stirred filtration dryer. Wash the filter cake with water-2 and -3 (5.0 L / kg x 2 times, stir for 15 minutes each wash) and isopropyl acetate-2 and 3 (5.0 L / kg x 2 times, stir for 15 min each wash). Dry the filter cake under nitrogen for 12 hours.

[0218] Acetone Repulping (Optional):

[0219] Charge pipazoline (Compound 6) and acetone (10.0 L / kg) into vessel E. Heat the suspension to 50 °C for 2 hours. Charge water-4 (10.0 L / kg) into vessel E over 1 hour. After completion of the water addition, cool the mixture to 20 °C over 1 hour. Filter the contents of vessel E to separate the product and wash the filter cake with a 1:1 acetone / water mixture (5.0 L / kg). Dry the filter cake under nitrogen for 12 hours.

[0220] Step 6

[0221]

[0222] General Note: All equivalents and volumes are referenced to the pipazoline input report

[0223]

[0224] Note: All L / kg and kg / kg amounts are relative to the pipazoline input

[0225] Reactor A was charged with a solution of pipazoline (Compound 6, 1.0 eq), degassed 2-MeTHF (9.0 L / kg), and potassium acetate (2.0 eq) 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)Cl 2 (0.003 eq) in 2-MeTHF (0.5 L / kg). Within 2 h of catalyst charge, over a period of >1 h but <2 h, a freshly prepared solution of cycloboroxane (Compound 6A, 0.5 eq) in wet degassed 2-MeTHF (4.0 L / kg, KF > 4.0%) was charged. After addition was complete, it was rinsed with an additional portion of wet 2-MeTHF (0.5 L / kg). After the reaction was complete (<0.15 area% of pipazoline remaining, typically <1 h after cycloboroxane addition was complete), 0.2 wt% (0.002 kg / kg) of biaryl seeds was added as a slurry in 0.02 L / kg of wet 2-MeTHF, and the resulting seed bed was aged for >60 min. At 75 °C ± 5 °C, heptane (5.0 L / kg) was added over 2 h. Then the batch was cooled to 20 °C ± 5 °C over 2 h and aged for another 2 h. Then the slurry was filtered, and the filter cake was washed with 1 x 5.0 L / kg of water, 1 x 5.0 L / kg of 1:1 iPrOH:water, and then 1 x 5.0 L / kg of 1:1 iPrOH:heptane (reslurry wash: resuspend the filter cake with a stirrer and allow to stand, then filter). Then the filter cake (biaryl, Compound 7) was dried by purging with nitrogen under vacuum.

[0226] Note: If the reaction stops, catalyst and cycloboroxane need to be recharged.

[0227] Step 7, Charcoal filtration for Pd removal

[0228]

[0229] General Note: All equivalents and volumes are referenced to the crude biaryl input report

[0230]

[0231] Note: All L / kg and kg / kg amounts are relative to the crude biaryl input

[0232] In a clean vessel A, crude biaryl (1 eq) was charged, and DCM (10 L / kg) was charged. At 22 °C ± 5 °C, the contents were stirred for >60 min and dissolution was observed. The crude biaryl from vessel A was added at ≤3 L 2The flux of / min / m passes through the bag filter and the carbon filter, and the filtrate is collected in the clean container B. The DCM rinse (1 L / kg) is charged into container A and collected in container B through the carbon filter.

[0233] Take a solution sample from the filtrate in container B for IPC Pd content. Concentrate the sample to a solid and analyze it by ICP-MS. IPC: Pd ≤ 25 ppm, relative to the biaryl.

[0234] a. If, relative to the biaryl, the Pd content is greater than 25 ppm in the first or second IPC sample, then pass the solution through the carbon filter for a second time at ≤ 3 L 2 / min / m 2 and rinse with 1 L / kg DCM; the sample filtrate is used for IPC.

[0235] b. If, after the third IPC, the Pd content is still greater than 25 ppm, then install and adjust a fresh carbon disk. Pass the biaryl filtrate through the updated carbon filter and wash with 1 L / kg DCM. The sample is used for IPC.

[0236] Distill and refill to the appropriate concentration. Prepare the distillation of the filtrate for recovery by concentrating to ≤ 4 L / kg DCM and refill to reach 5.25 ± 0.25 L / kg DCM, then move to step 7 Boc-deprotection reaction.

[0237] Step 7

[0238]

[0239] General Note: All equivalents and volumes are referenced to the crude biaryl input report

[0240]

[0241]

[0242] Note: All L / kg and kg / kg amounts are relative to the biaryl input

[0243] Add to reactor A: 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]pyrimidine-4-yl}-3-methylpiperazine-1-carboxylate (biaryl) (1.0 equivalent), dichloromethane (5.0 L / kg), and slowly charge TFA (15.0 equivalents, 1.9 L / kg) to maintain the internal temperature at 20 °C ± 5 °C. Stir the reaction at 20 °C ± 5 °C for 4 h.

[0244] Add to Reactor B: 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 ± 5 °C for an additional 12 h.

[0245] Filter the resulting slurry and wash with water (2 x 10 L / kg). Dry the wet filter cake for 24 h to obtain 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperazin-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).

[0246] Step 8

[0247]

[0248] General Note: All equivalents and volumes are referenced to the Des-Boc input report

[0249]

[0250] Note: All L / kg and kg / kg amounts are relative to the Des-Boc input

[0251] Under nitrogen, charge Des-Boc (Compound 8, 1.0 equivalent) and NMP (4.2 L / kg) to vessel A. Slowly charge TFA (1.0 equivalent) to maintain Tr < 25 °C. Age the mixture at 25 °C until complete dissolution is observed (approximately 0.5 h). Then finely filter the solution through a 0.45 micron filter into vessel B and wash with NMP (0.8 L / kg). Combine the filtrate and washings, then cool to 0 °C. Add acryloyl chloride (1.3 equivalents) to the resulting solution while maintaining the temperature < 10 °C. Then age the reaction mixture at 5 °C ± 5 °C until completion by IPC (approximately 1.5 h).

[0252] Preparation of sodium phosphate aqueous quench solution:

[0253] Charge sodium phosphate (3.0 equivalents) and water (15.0 L / kg) to vessel C. Age the mixture at 25 °C until complete dissolution is observed. Warm the solution to 45 °C ± 5 °C. Prepare a seed slurry of AMG 510 (0.005 equivalent) in water (0.4 L / kg) and add it to vessel C while maintaining the temperature at 45 °C ± 5 °C.

[0254] Transfer the reaction mixture in vessel B to vessel C (quenching solution) while maintaining the temperature at 45 °C ± 5 °C (for about 1 hour). Wash vessel B with a portion of NMP (0.5 L / kg). Age the product slurry at 45 °C ± 5 °C for 2 hours, cool to 20 °C over 3 hours, age at 20 °C for at least 12 hours, filter and wash with water (2 x 10.0 L / kg). Dry the product using nitrogen and vacuum to obtain crude AMG 510 (Compound 9A).

[0255] Step 9

[0256]

[0257] General Note: All equivalents and volumes are referenced to the crude AMG 510 input report

[0258]

[0259] Note: All L / kg and kg / kg amounts are relative to the crude AMG 510 input

[0260] Reactor A is charged with 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (crude AMG 510) (1.0 equivalent), ethanol (7.5 L / kg), and water (1.9 L / kg). Heat the mixture to 75 °C and filter finely into a clean reactor B. Cool the solution to 45 °C and inoculate with authentically milled AMG 510 seeds (0.015 ± 0.005 kg / kg); age the resulting slurry for 30 min. Add water (15.0 L / kg) over 5 h while maintaining the internal temperature > 40 °C; age the mixture for an additional 2 h.

[0261] Cool the mixture to 20 °C over 3 hours and age for 8 h, then collect the solid by filtration and wash with a mixture of ethanol (2.5 L / kg) and water (5.0 L / kg). Dry the solid using vacuum and nitrogen to obtain 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (AMG 510, Compound 9).

[0262] Compound 6A Cycloboroxane Synthesis:

[0263] Lithiation / Boration

[0264]

[0265]

[0266] Reactor A was charged with THF (6 volumes), a secondary amine base, diisopropylamine (1.4 equivalents), and a catalyst such as triethylamine hydrochloride (0.01 equivalent). The resulting solution was cooled to -70 °C, and the first base n-BuLi (2.5 M in hexanes, 1.5 equivalents) was added slowly. After the addition was complete, a solution of 3-fluoroanisole (1.0 equivalent) in THF (6 volumes) was added slowly and held at -70 °C for 5 min. Reagent B (EtO) 3 (2.0 equivalents) was added slowly either simultaneously or sequentially and held at -70 °C for 10 min. The reaction mixture was quenched with an acid, 2 N HCl. The quenched reaction mixture was extracted with MTBE (3 × 4 volumes). The combined organic phases were concentrated to 1.5 - 3 total volumes. Heptane (7 - 9 volumes) was added dropwise, and the mixture was cooled to 0 °C - 10 °C and stirred for 3 h. The mixture was filtered and rinsed with heptane (1.5 volumes). The solid was dried under nitrogen at <30 °C to give (2-fluoro-6-methoxyphenyl)boronic acid.

[0267] Demethylation:

[0268]

[0269]

[0270]

[0271] Note: All L / kg and kg / kg amounts are relative to the (2-fluoro-6-methoxyphenyl)boronic acid input

[0272] The reactor was charged with dichloromethane (solvent, 4.0 L / kg) and an acid, BBr 3 (1.2 equivalents), and cooled to -20 °C. To this solution, a suspension of (2-fluoro-6-methoxyphenyl)boronic acid (1.0 equivalent) in dichloromethane (4.0 L / kg) was added to BBr 3In the / DCM mixture, while maintaining the temperature at -15°C to -25°C. Allow the reaction to proceed for about 2 hours while monitoring by HPLC [≤1% (2-fluoro-6-methoxyphenyl)boronic acid], and then quench it back into water (3.0 L / kg). Then the precipitated solid is separated by filtration and slurried with water (3.0 L / kg) on the filter, and then de-liquored. The filtrate is adjusted to pH 4 - 6 by adding sodium bicarbonate. The bottom organic phase is separated, and the resulting aqueous layer is washed with dichloromethane (solvent, 5.0 volumes) and adjusted to pH = 1 by adding concentrated hydrochloric acid. The resulting solid is separated by filtration, and the filter cake is washed with water (2x5.0 L / kg).

[0273] Purification by re-slurrying (required)

[0274] Charge the combined crude solid into a reactor and slurry it with 5% EtOH / water (5.0 L / kg) at 20°C for >1 h. Then the purified product is separated by filtration and rinsed with water (2x3 L / kg), and then dried with nitrogen / vacuum on the filter at <30°C to obtain 2,2',2''-(1,3,5,2,4,6-trioxatriborane-2,4,6-triyl)tris(3-fluorophenol) (cyclic boroxine, Compound 6A).

[0275] The foregoing is only illustrative of the invention and is not intended to limit the invention to the disclosed uses. Variations and changes 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 mentioned are hereby incorporated by reference in their entirety as if written herein.

Claims

1. A compound having formula 6A:

2. A composition comprising a compound having formula 6A:

3. A method for preparing a compound having formula 6A: The method comprises mixing (2-fluoro-6-methoxyphenyl)boronic acid with an acid in a solvent to form a compound having formula 6A.

4. The method according to claim 3, wherein the acid is BBr 3 .

5. The method according to claim 3, wherein the at least one solvent is dichloromethane.

6. The method according to claim 3, wherein the at least one solvent is heptane.

7. The method according to claim 3, wherein the mixture is cooled to about -20 °C.

8. The method according to claim 3, wherein the method for preparing a compound having the following structure: comprises mixing a compound having the following structure: with triethyl borate, a first base, a secondary amine base, a catalyst, and an acid.

9. The method according to claim 8, wherein the first base is n-butyllithium.

10. The method according to claim 8, wherein the secondary amine base is diisopropylamine.

11. The method according to claim 8, wherein the catalyst is triethylamine hydrochloride.

12. The method according to claim 8, wherein the acid is HCl.

13. A method for preparing a compound having formula 7, The method comprises the steps of: reacting a compound having formula 6A: with a compound having formula 6: React in the presence of Pd(dpePhos)Cl 2 and KOAc.

Images