Improved synthesis method for KRAS G12C inhibitor compounds

Abstract

This invention provides an improved, efficient, and scalable method for preparing intermediate compounds, such as 2-isopropyl-4-methylpyridine-3-amine, which are useful for the synthesis of compounds for the treatment of KRAS G12C-mutated cancers. [Solution] Structure JPEG2026102821000066.jpg26153 A method for producing the following compounds is provided, comprising reacting a mixture containing a compound with palladium in a solvent in the presence of hydrogen. JPEG2026102821000067.jpg21153

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 935,515, filed November 14, 2019, which is hereby incorporated by reference in its entirety.

[0002] The present disclosure relates to an improved, efficient, and scalable method for preparing intermediate compounds, such as compounds of Formula 1B, useful in the synthesis of compounds for the treatment of KRAS G12C mutant cancers, having the structure

Chemical Formula

Background Art

[0003] 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 in some studies, have been shown to be a negative prognostic factor for NSCLC patients. Recently, V - Ki - ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations have been found to confer resistance to epidermal growth factor receptor (EGFR) targeted therapy in colorectal cancer, and thus, the mutational status of KRAS can provide important information prior to the prescription of TKI therapy. In summary, there is a need for new medical treatments for patients with pancreatic cancer, lung adenocarcinoma, or colorectal cancer, particularly patients diagnosed with cancers characterized by KRAS mutations, and patients who have progressed even after chemotherapy.

Summary of the Invention

Means for Solving the Problems

[0004] The present disclosure provides the following chemical structure:

Chemical Formula

Mode for Carrying Out the Invention

[0005] Definitions Abbreviations: The following abbreviations may be used in this specification:

[0006]

Table 1

[0007]

Table 2

[0008]

Table 3

[0009] The use of the terms "a", "an", "the", and similar referents in the context of this disclosure (especially in the context of the claims) should be construed to include both the singular and the plural unless otherwise indicated. The recitation of a range of values herein is merely intended to serve as a concise way of referring individually to each separate value falling within that range, and each separate value is incorporated herein as if it were individually recited herein. The use of any examples, or exemplary language (e.g., "such as") provided herein is intended to better illustrate the disclosure and is not intended to limit the scope of the disclosure unless otherwise specifically mentioned. No language in this specification should be construed as indicating that any non-recited element is essential for the implementation of the disclosure.

[0010] As used herein, the term “alkyl” refers to linear and branched C1-C8 hydrocarbon groups, including, but not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, t-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. m~n This means that the alkyl group has m to n carbon atoms. The term "alkylene" refers to alkyl groups with substituents. Alkyl (e.g., methyl) or alkylene (e.g., -CH2-) groups include, for example, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, and C. 1~8 Alkyl, C 2~8 Alkenil, C 2~8 Alkinyl, -NC, amino, -CO2H, -CO2C1~C8 alkyl, -OCOC1~C8 alkyl, C3~C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5~C 10 Aryl, and C5~C 10 One or more, usually 1 to 3, atoms can be substituted independently of heteroaryls. The term "haloalkyl" specifically refers to an alkyl group in which at least one, e.g., 1 to 6, or all of the hydrogen atoms of the alkyl group are substituted with halo atoms.

[0011] The terms "alkenyl" and "alkynyl" refer to alkyl groups that further contain double or triple bonds, respectively.

[0012] As used herein, the term "halo" refers to fluoro, chloro, bromo, and iodine. The term "alkoxy" is defined as -OR (where R is alkyl).

[0013] As used herein, the terms "amino" or "amine" refer interchangeably to a -NR2 group, where each R is, for example, H or a substituent. In some embodiments, the amino group is further substituted to form an ammonium ion, such as NR3 + is formed. Substituents that are specifically included in the definition of "ammonium" or "amine" can be, for example, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, amide, or carboxylate. The R groups can further be substituted with one or more, for example 1 to 4, groups selected from, for example, halo, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, carboxylate, amine, and amide. The term "amide" or "amide group" refers interchangeably to a group that is similar to an amine or amino group but further includes C(O), for example, -C(O)NR2.

[0014] 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, azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Aryl also refers here to bicyclic and tricyclic carbocyclic rings of C 10~14 where one ring is an aromatic ring and the other is a saturated ring, a partially unsaturated ring, or an aromatic ring, such as dihydronaphthyl, indenyl, indanyl, or tetrahydronaphthyl (tetralinyl). Unless otherwise specified, an aryl group is, for example, halo, C 1-8 alkyl, C 2~8 alkenyl, C 2~8 alkynyl, -CF3, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 cycloalkyl, C3-C 10 heterocycloalkyl, C5-C10 Aryl, and C5~C 10 It may be substituted with one or more groups, particularly 1 to 4 groups, independently selected from the heteroaryl.

[0015] As used herein, the term "cycloalkyl" refers to monocyclic or polycyclic non-aromatic carbocyclic rings (where the polycyclic ring may be condensed, cross-linked, or spiro). A carbocyclic ring may have 3 to 10 carbocyclic atoms. Possible carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclononyl.

[0016] As used herein, the term "heterocycloalkyl" means a ring system containing three or more (e.g., 3 to 12, 4 to 10, 4 to 8, or 5 to 7) total atoms, of which 1 to 5 (e.g., 1, 2, 3, 4, or 5) atoms are monocyclic or polycyclic (e.g., bicyclic) saturated or partially unsaturated rings independently selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heterocycloalkyl groups include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, dihydropyrrolyl, morpholinyl, thiomorpholinyl, dihydropyridinyl, oxacycloheptyl, dioxacycloheptyl, thiacycloheptyl, and diazacycloheptyl.

[0017] Unless otherwise specified, cycloalkyl groups or heterocycloalkyl groups may be unsubstituted or substituted with one or more groups, particularly 1 to 4 groups. Some possible substituents include halo, C 1~8 Alkyl, C 2~8 Alkenil, C 2~8 Alkinyl, -OCF3, -NO2, -CN, -NC, -OH, Alkoxy, Amino, -CO2H, -CO2C1~C8 alkyl, -OCOC1~C8 alkyl, C3~C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5~C 10 Aryl, and C5~C 10Heteroaryls are one example.

[0018] As used herein, the term “heteroaryl” refers to a monocyclic or polycyclic (e.g., bicyclic) ring system comprising 1 to 3 aromatic rings, the aromatic rings containing 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from nitrogen, oxygen, and sulfur. In certain embodiments, the heteroaryl group has 5 to 20, 5 to 15, 5 to 10, or 5 to 7 atoms. A heteroaryl is also a ring in which one ring is aromatic and the other is saturated, partially unsaturated, or aromatic. 10~14 This refers to bicyclic and tricyclic rings. Examples of heteroaryl groups include furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridadinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl, benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzothiophenyl, and benzotriazoli Examples of heteroaryl groups include, but are not limited to, halo, benzoxazolyl, flopyridyl, imidazopyridinyl, imidazothiazolyl, indolidinyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, chiazolinyl, thiadiazolopyrimidyl, and thienopyridyl. Unless otherwise specified, heteroaryl groups may be unsubstituted or substituted with one or more substituents, particularly 1 to 4 or 1 or 2 substituents. Possible substituents include halo, C 1~8 Alkyl, C 2~8 Alkenil, C 2~8 Alkinyl, -OCF3, -NO2, -CN, -NC, -OH, Alkoxy, Amino, -CO2H, -CO2C1~C8 alkyl, -OCOC1~C8 alkyl, C3~C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5~C10 Aryl, and C5~C 10 Heteroaryls are one example.

[0019] As used herein, the term Boc refers to the following structure: [ka]

[0020] Embodiment Embodiment 1 In one embodiment of this disclosure, this disclosure relates to formula 2A: [ka] A method for producing the compound, wherein the structure is: [ka] The method includes reacting a mixture containing a compound with palladium in a solvent in the presence of hydrogen.

[0021] Embodiment 2 In another embodiment of the present disclosure, the present disclosure is obtained by reacting a mixture comprising 1,2-dimethoxyethane (DME), an aqueous solution of K2CO3, Pd(PPh3)Cl2, 2-chloro-4-methylpyridine-3-amine, and 4,4,5,5-tetramethyl-2-(propa-1-en-2-yl)-1,3,2-dioxaborolane to produce a structure: [ka] The method of Embodiment 1 includes producing a compound having [a specific characteristic].

[0022] Embodiment 3 In another embodiment of this disclosure, this disclosure uses formula 2A: [ka] A method for producing the compound, comprising an aqueous solution of NaOH, structure: [ka] The method includes reacting a compound 36 having a certain property with a mixture containing sodium hypochlorite.

[0023] Embodiment 4 In another embodiment of this disclosure, the structure: [ka] The method of Embodiment 3 includes producing compound 36 by reacting compound 35 having with H2SO4.

[0024] Embodiment 5 In another embodiment of this disclosure, the structure: [ka] Compound 34 having the following structure is reacted with DMF-DMA, NH4OH, and NH4OOCCH3 to obtain a compound with the following structure: [ka] The method of Embodiment 4 includes producing a compound 35 having [a certain characteristic].

[0025] Embodiment 6 In another embodiment of this disclosure, the structure: [ka] Compound 33 having the following structure is reacted with potassium tert-butoxide, ethyl isobutyrate, acetonitrile and L-proline to obtain a compound with the following structure: [ka] The method of Embodiment 5 includes producing a compound 34 having the following characteristics.

[0026] Embodiment 7 In another embodiment of this disclosure, the structure: [ka] A method for producing a compound 35 having the following characteristics: a.) Crotonaldehyde and (S)-α,α-bis[3,5-bis(trifluoromethyl)phenyl]-2-pyrrolidinemethanoltrimethylsilyl ether; b.) 4-methyl-3-oxopentannitrile; c.) Acetonitrile; and d.) Hydroxylamine hydrochloride The method includes a step of reacting a mixture, which includes a step of reacting an ingredient.

[0027] Embodiment 8 In another embodiment of the present disclosure, the present disclosure uses the compound of formula 2A to represent formula 9: [ka] The method of Embodiment 1 includes producing a compound having [a specific characteristic].

[0028] Embodiment 9 In another embodiment of the present disclosure, the present disclosure uses the compound of formula 2A to represent formula 9: [ka] The method of Embodiment 3 includes producing a compound having [a specific characteristic].

[0029] Embodiment 10 In another embodiment of the present disclosure, the present disclosure uses the compound of formula 2A to represent formula 9: [ka] The method of Embodiment 7 includes producing a compound having [a specific characteristic].

[0030] Embodiment 11 In another embodiment of the present disclosure, the present disclosure includes the method of Embodiment 10, further comprising mixing a compound of Formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

[0031] Embodiment 12 In another embodiment of the present disclosure, the present disclosure includes the method of Embodiment 11, further comprising mixing a compound of Formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

[0032] Embodiment 13 In another embodiment of the present disclosure, the present disclosure includes the method of Embodiment 12, further comprising mixing a compound of Formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

[0033] Compounds of the Disclosure This specification provides KRAS inhibitors having a structure described in more detail below.

[0034] The compounds disclosed herein include all pharmaceutically acceptable isotope-labeled compounds in which one or more atoms of the disclosed compound are substituted with atoms having the same atomic number but with an atomic mass or mass number different from those commonly 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, for example, 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, 36Cl, 123 I, and 125 I is an example. These radiolabeled compounds may be useful for helping to determine or measure the efficacy 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 isotope-labeled compounds of this disclosure, for example, compounds incorporating radioisotopes, are useful in studies of drug and / or substrate tissue distribution. Radioisotopes such as tritium, i.e. 3 H, and carbon-14, i.e. 14 C is particularly useful for this purpose in terms of the ease of their incorporation and the ease of detection.

[0035] Deuterium, that is 2 Substitution with heavier isotopes, such as 1H, can provide greater metabolic stability, offering certain therapeutic benefits resulting from, for example, an increased in vivo half-life or reduced drug requirements, and is therefore preferable in some situations.

[0036] 11 C, 18 F, 15 O and 13 Substitution with positron-emitting isotopes such as 1N may be useful in positron-emitting topography (PET) studies to investigate substrate-receptor occupancy. The isotope-labeled compound of structure (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described below, using appropriate isotope-labeling reagents instead of previously used unlabeled reagents.

[0037] The isotope-labeled compounds disclosed herein can generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described in the accompanying examples and schemes, using appropriate isotope-labeled reagents instead of previously used unlabeled reagents.

[0038] Some of the compounds disclosed herein may exist as stereoisomers (i.e., isomers that differ only in the spatial arrangement of atoms), including optical isomers and conformational isomers (or conformers). The compounds disclosed herein include all stereoisomers as pure individual stereoisomer preparations, as each as a Rich preparation, as a racemic mixture of such stereoisomers, or as individual diastereomers and enantiomers that can be separated according to methods known to those skilled in the art. Furthermore, the compounds disclosed herein include all tautomers of those compounds.

[0039] Some of the compounds disclosed herein may exist as atropisomers, which are conformational isomers that arise when rotation around a single bond in a molecule is hindered or very slowed as a result of steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers as pure individual atropisomer preparations, as Richie preparations for each, or as nonspecific mixtures thereof. Separation and isolation of isomer species may be possible if the rotational barrier around the single bond is sufficiently high and the interconversion between conformations is sufficiently slow. For example, the following groups may restrict rotation: [ka]

[0040] The term "monohydrate" refers to a salt of compound 9 to which approximately one water molecule is bonded. Those skilled in the art will understand that the exact number of bonded water molecules can change slightly at any time as temperature, pressure, and other environmental influences change. All slight variations in the number of bonded water molecules are considered to be within the scope of this disclosure.

[0041] The term “dihydrate” refers to a salt of compound 9 in which approximately two water molecules are bonded. Those skilled in the art will understand that the exact number of bonded water molecules can change slightly at any time as temperature, pressure, and other environmental influences change. All slight changes in the number of bonded water molecules are considered to be within the scope of this disclosure.

[0042] The term "cocrystal" refers to a crystalline material containing two or more compounds at ambient temperature (20°C to 25°C, preferably 20°C), where at least two of the compounds are bonded by weak interactions, at least one of the compounds is a cocrystal-forming agent, and the others are compound 5. Weak interactions are defined as interactions that are neither ionic nor covalent, such as hydrogen bonds, van der Waals forces, and π-π interactions.

[0043] The term "amorphous morphology" or "amorphous" refers to a material that lacks long-range order and does not exhibit a distinct X-ray diffraction peak, i.e., a Bragg diffraction peak. The XRPD pattern of an amorphous material is characterized by one or more amorphous halos.

[0044] The term "amorphous halo" refers to a nearly bell-shaped maximum in the X-ray powder pattern of an amorphous material.

[0045] The term "substantially pure" refers to the solid form of compound 9 having a purity of approximately 95%, particularly approximately 99.5%, more particularly approximately 99.8%, and even more particularly approximately 99.9%.

[0046] The term "patient" refers to animals such as dogs, cats, cows, horses, sheep, and humans. Certain patients are mammals. The term "patient" includes both males and females.

[0047] Terms such as "treating," "treating," or "treatment" include preventative (e.g., prophylactic) and palliative treatments.

[0048] The term "excipients" refers to pharmaceutically acceptable additives, carriers, diluents, adjuvants, or other components other than the active pharmaceutical ingredient (API), which are typically included for formulation and / or administration to a patient.

[0049] Pharmaceutical composition, dosage, and route of administration Furthermore, provided herein are pharmaceutical compositions comprising the compounds disclosed herein together with pharmaceutically acceptable excipients, such as diluents or carriers. Compounds and pharmaceutical compositions suitable for use in this disclosure include those in which the compound can be administered in an effective amount to achieve its intended purpose. The administration of the compounds is described in more detail below.

[0050] Those skilled in the art can determine the appropriate pharmaceutical dosage form depending on the route of administration and the desired dose. See, for example, Remington's Pharmaceutical Sciences, 1435-712 (18th ed., Mack Publishing Co, Easton, Pennsylvania, 1990). The dosage form may affect the physical state, stability, in vivo release rate, and in vivo elimination rate of the administered drug. Depending on the route of administration, the appropriate dose may be calculated according to body weight, body surface area, or organ size. Further refinements to the calculations necessary to determine the appropriate therapeutic dose are routinely made by those skilled in the art without excessive experimentation, particularly in light of the dose information and analyses disclosed herein, as well as pharmacokinetic data obtained through animal or human clinical trials.

[0051] The terms "pharmaceutically acceptable" or "pharmacologically acceptable" refer to molecular entities and compositions that, when administered to animals or humans, do not produce harmful allergic or other adverse reactions. In this specification, "pharmaceutically acceptable" includes any and all solvents, dispersion media, coatings, antimicrobial and antifungal agents, isotonic agents and absorption retarders, etc. The use of such excipients for pharmaceutically active substances is well known in the art. Any conventional media or agent is intended for use in a therapeutic composition unless it is incompatible with the therapeutic composition. Auxiliary active ingredients may also be incorporated into the composition. In exemplary embodiments, the formulation includes corn syrup solid, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tricalcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dipotassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, It may contain L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, tocopherol mixture, calcium pantothenate, copper sulfate, thiamine hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, β-carotene, potassium iodide, phylloquinone biotin, sodium selenite, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0052] The compounds may be present in pharmaceutical compositions as pharmaceutically acceptable salts. In this specification, “pharmaceutically acceptable salts” include, for example, base addition salts and acid addition salts.

[0053] pharmaceutically acceptable base addition salts can be produced using metals or amines such as alkalis and alkaline earth metals or organic amines. pharmaceutically acceptable salts of compounds can also be prepared using pharmaceutically acceptable cations. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkali, alkaline earth, ammonium, and quaternary ammonium cations. Carbonates or bicarbonates are also possible. Examples of metals used as cations include sodium, potassium, magnesium, ammonium, calcium, or ferric. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, procaine chloride, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine.

[0054] Pharmaceutically acceptable acid addition salts include inorganic or organic acid salts. Examples of suitable acid salts include hydrochloride, formate, acetate, citrate, salicylate, nitrate, and 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; organic carboxylic acids, sulfonic acids, sulfoacids, or phosphoacids or N-substituted sulfamic acids, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, and ni This includes substances containing chotinic acid or isonicotinic acid; and substances containing 20 alpha amino acids involved in the synthesis of natural proteins, such as glutamic acid or aspartic acid; and also substances containing 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- or 3-phosphoglyceric acid, glucose 6-phosphate, N-cyclohexylsulfamic acid (with cyclamate formation), or other acidic organic compounds such as ascorbic acid.

[0055] Pharmaceutical compositions containing the compounds disclosed herein can be manufactured by conventional methods, for example, by conventional mixing, dissolution, granulation, sugar-coated tablet manufacturing, powdering, emulsification, encapsulation, encapsulation, or lyophilization processes. The appropriate dosage form depends on the chosen route of administration.

[0056] For oral administration, suitable compositions can be readily formed by combining the compounds disclosed herein with pharmaceutically acceptable excipients, such as carriers well known in the art. Such excipients and carriers enable the compounds disclosed herein to be formed into dosage forms such as tablets, pills, coated tablets, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral intake by patients being treated. Pharmaceutical preparations for oral use can be obtained by adding a solid excipient to the compounds disclosed herein, optionally grinding the resulting mixture, and, if necessary, adding suitable adjuvants to obtain a tablet or coated tablet core, and then processing the granular mixture. Suitable excipients include, for example, fillers and cellulose preparations. Disintegrants may be added as needed. Pharmaceutically acceptable ingredients are well known in various types of formulations and may include, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavorings, coating materials, preservatives, dyes, thickeners, adjuvants, antimicrobial agents, antioxidants, and carriers for various formulation types.

[0057] When a therapeutically effective dose of the compounds disclosed herein is administered orally, the compositions are typically in the form of solids (e.g., tablets, capsules, pills, powders, or lozenges) or liquid formulations (e.g., aqueous suspensions, solutions, elixirs, or syrups).

[0058] When administered in tablet form, the composition may further contain a functional solid and / or solid carrier, such as gelatin or an adjuvant. Tablets, capsules, and powders may contain about 1 to about 95% of the compound, preferably about 15 to about 90%.

[0059] When administered in liquid or suspension form, functional liquids and / or liquid carriers, such as water, petroleum, or oils of animal or plant origin, may be added. The liquid form of the composition may further contain physiological saline, sugar alcohol solutions, dextrose or other sugar solutions, or glycols. When administered in liquid or suspension form, the composition may contain about 0.5 to about 90% by weight of the compounds disclosed herein, preferably about 1 to about 50% of the compounds disclosed herein. In one possible embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. When administered in liquid form, the composition may be supplied as a rapidly dissolving solid formulation for dissolution or suspension immediately before administration.

[0060] When a therapeutically effective dose of the compounds disclosed herein is administered intravenously, cutaneously, or subcutaneously, the composition is in the form of a parenterally acceptable aqueous solution that does not contain pyrogens. Preparing such parenterally acceptable solutions, taking into account pH, isotonicity, stability, etc., is within the scope of the art. Preferred compositions for intravenous, cutaneous, or subcutaneous injection typically contain an isotonic vehicle in addition to the compounds disclosed herein. Such compositions may be prepared for administration as a solution of a free base or a pharmacokinetically acceptable salt in water, appropriately mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as in oil. Under normal storage and use conditions, these preparations may optionally contain preservatives to prevent microbial growth.

[0061] The injectable composition may include sterile aqueous solutions, suspensions, or dispersions, and sterile powders for the immediate preparation of sterile injectable solutions, suspensions, or dispersions. In all embodiments, this form must be sterile and fluid enough to pass through an injection needle easily. It must be stable under manufacturing and storage conditions and resistant to microbial contamination, such as bacteria and fungi, by optionally including preservatives. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils. In one possible embodiment, the carrier is non-aqueous or substantially non-aqueous. Adequate fluidity can be maintained, for example, by the use of a coating such as lecithin, maintaining the required particle size of the compound in the dispersion embodiment, and the use of surfactants. Prevention of microbial activity can be provided by various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many embodiments, it is preferable to include an isotonic agent, such as sugar or sodium chloride. The extension of absorption of an injectable composition can be achieved by using absorption-delaying agents, such as aluminum monostearate and gelatin, in the composition.

[0062] Sterile injectable solutions are prepared by incorporating the required amount of active compound, along with various other components listed above if necessary, into a suitable solvent, followed by sterilization by filtration. Generally, dispersions are prepared by incorporating various sterile active ingredients into a sterile vehicle containing the basic dispersion medium and other necessary components listed above. In embodiments of sterile powders for preparing sterile injectable solutions, preferred preparation methods are vacuum drying and freeze-drying, in which the powders of the active ingredients and any additional desired ingredients are obtained from their pre-sterilized filtered solutions.

[0063] Slow-release formulations, i.e., sustained-release formulations, can also be prepared to achieve controlled release of the active compound in contact with body fluids within the GI tube, thereby providing a substantially constant and effective plasma concentration of the active compound. For example, release can be controlled by one or more of the following: dissolution, diffusion, and ion exchange. Furthermore, the sustained-release approach can enhance absorption via saturable or restricted pathways within the GI tube. For example, the compound can be embedded for this purpose in a matrix of biodegradable polymers, water-soluble polymers, or mixtures thereof, and optionally in a suitable surfactant. Embedding in this context means incorporating microparticles into a polymer matrix. Controlled-release formulations can also be obtained by encapsulating dispersed microparticles or emulsified microdroplets using well-known dispersion or emulsification coating techniques.

[0064] For administration by inhalation, the compounds of this disclosure are conveniently delivered in the form of an aerosol spray from a pressurized container or nebulizer using a suitable propellant. In the pressurized aerosol embodiment, the dose unit can be determined by providing a valve for delivering a measured amount. Capsules and cartridges, for example, gelatin capsules and cartridges for use in inhalers or injectors, can be formulated to contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0065] The compounds disclosed herein can be formulated for parenteral administration by injection (e.g., bolus injection or serial infusion). Injectable formulations can be provided in unit dose form (e.g., ampoules or multi-dose containers) with added preservatives. Compositions can take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles and may contain formulation agents such as suspending agents, stabilizers, and / or dispersants.

[0066] Pharmaceutical formulations for parenteral administration contain aqueous solutions of water-soluble compounds. Furthermore, suspensions of compounds can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. Optionally, suspensions may also contain suitable stabilizers or agents that increase the solubility of the compound and enable the preparation of highly concentrated solutions. Alternatively, the composition may be in powder form for preparation with a suitable vehicle (e.g., sterile, pyrogen-free water) before use.

[0067] The compounds disclosed herein can also be formulated into rectal compositions such as suppositories or retaining enemas (e.g., including conventional suppository bases). In addition to the formulations described above, the compounds can also be formulated as depot formulations. Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Accordingly, for example, the compounds can be formulated with a suitable polymer or hydrophobic material (e.g., as an emulsion in an acceptable oil), or with an ion exchange resin, or as a sparingly soluble derivative, for example, as a sparingly soluble salt.

[0068] In particular, the compounds disclosed herein may be administered orally, orally, or sublingually, in the form of tablets containing excipients such as starch or lactose, or in the form of capsules or ovules, alone or in mixtures with excipients, or in the form of elixirs or suspensions containing flavoring agents or coloring agents. Such liquid preparations may be prepared with pharmaceutically acceptable additives such as suspensions. The compounds may also be administered parenterally, for example, by injection intravenously, intramuscularly, subcutaneously, or into the coronary artery. For parenteral administration, the compounds are most often used in the form of sterile aqueous solutions that may contain other substances, such as salts, or sugar alcohols such as mannitol, or glucose, in order to make the solution isotonic with blood.

[0069] For veterinary use, the compounds disclosed herein are administered as appropriately acceptable formulations in accordance with normal veterinary practice. Veterinarians can easily determine the most appropriate administration plan and route for a particular animal.

[0070] In some embodiments, all components necessary for the treatment of KRAS-related disorders, either used alone or in combination with other agents or therapeutic interventions traditionally used for the treatment of such disorders, may be packaged in a kit. Specifically, this disclosure provides a kit for use in a therapeutic intervention for a disorder, comprising a packaged set of agents including the compounds disclosed herein, buffers and other components for preparing the agents into a deliverable form, and / or a device for delivering such agents, and / or any agents used in combination therapy with the compounds disclosed herein, and / or instructions for the treatment of the disorder packaged together with the agents. The instructions may be fixed on printed paper or any tangible medium such as computer-readable magnetic or optical media, or the instructions may refer to a data source on a remote computer, such as a World Wide Web page accessible via the Internet.

[0071] “Therapeutic dose” means an amount that is effective in treating, slowing the progression of, or alleviating the current symptoms of the subject being treated. Determining the effective dose is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, “therapeutic dose” refers to the amount of compound that results in the achievement of the desired effect. For example, in one preferred embodiment, a therapeutic dose of a compound disclosed herein reduces KRAS activity by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% compared to a control.

[0072] The amount of compound administered may depend on the subject being treated, their age, health, sex, and weight, the type of concomitant therapy (if any), the severity of the distress, the nature of the desired effect, the method and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on arterial oxygen pressure. However, the most preferred dose can be determined to suit the individual subject without excessive experimentation, as can be understood and determined by those skilled in the art. This usually involves adjusting the standard dose (e.g., reducing the dose if the patient is underweight).

[0073] While individual needs vary, determining the optimal range of effective amounts of the compounds is within the scope of those skilled in 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 disclosed herein can be about 0.05 mg / kg / day to about 50 mg / kg / day, for example, 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, which may be, for example, about 2.5 mg / day (0.5 mg / kg × 5 kg) to about 5000 mg / day (50 mg / kg × 100 kg). For example, the dosage of the compound ranges from approximately 0.1 mg / kg / day to approximately 50 mg / kg / day, approximately 0.05 mg / kg / day to approximately 10 mg / kg / day, approximately 0.05 mg / kg / day to approximately 5 mg / kg / day, approximately 0.05 mg / kg / day to approximately 3 mg / kg / day, approximately 0.07 mg / kg / day to approximately 3 mg / kg / day, approximately 0.09 mg / kg / day to approximately 3 mg / kg / day, and approximately 0.05 mg / kg / day to approximately 0.1 mg / kg / day. The dosage can be approximately 0.1 mg / kg / day to approximately 1 mg / kg / day, approximately 1 mg / kg / day to approximately 10 mg / kg / day, approximately 1 mg / kg / day to approximately 5 mg / kg / day, approximately 1 mg / kg / day to approximately 3 mg / kg / day, approximately 3 mg / day to approximately 500 mg / day, approximately 5 mg / day to approximately 250 mg / day, approximately 10 mg / day to approximately 100 mg / day, approximately 3 mg / day to approximately 10 mg / day, or approximately 100 mg / day to approximately 250 mg / day. Such dosages may be administered as a single dose or divided into multiple doses.

[0074] How to use KRAS G12C inhibitors This disclosure provides a method for inhibiting RAS-mediated cell signaling, comprising contacting cells with an effective amount of one or more compounds disclosed herein. Inhibition of RAS-mediated signaling can be evaluated and demonstrated by a wide range of methods well known in the art. Non-limiting examples include (a) a decrease in RAS GTPase activity; (b) a decrease in GTP binding affinity or an increase in GDP binding affinity; (c) an increase in K-off of GTP or a decrease in K-off of GDP; (d) a decrease in the levels of downstream signaling molecules of the RAS pathway, such as a decrease in pMEK, pERK, or pAKT levels; and / or (e) a decrease in the binding of the RAS complex to downstream signaling molecules, including but not limited to Raf. Kits and commercially available assays can be utilized to determine one or more of the above.

[0075] The disclosure also provides methods of using the compounds or pharmaceutical compositions of the disclosure to treat disease conditions, including but not limited to, those associated with G12C KRAS, HRAS, or NRAS mutations (e.g., cancer).

[0076] In some embodiments, a method for treating cancer is provided, which comprises administering an effective amount of any of the aforementioned pharmaceutical compositions containing the compounds disclosed herein to a subject in need thereof. In some embodiments, the cancer is mediated by a G12C mutation in KRAS, HRAS, or NRAS. In various embodiments, the cancer is pancreatic cancer, colorectal cancer, or lung cancer. In some embodiments, the cancer is gallbladder cancer, thyroid cancer, and bile duct cancer.

[0077] In some embodiments, the Disclosure provides a method for treating a disorder in a subject requiring treatment, comprising determining whether the subject has a G12C mutation in KRAS, HRAS, or NRAS, and if it is determined that the subject has a G12C mutation in KRAS, HRAS, or NRAS, administering to the subject a therapeutically effective dose of at least one compound disclosed herein or a pharmaceutically acceptable salt thereof.

[0078] The disclosed compounds have the potential to inhibit anchorage-independent cell proliferation and therefore tumor metastasis. Accordingly, in another embodiment, the present disclosure provides a method for inhibiting tumor metastasis, the method comprising administering an effective amount of the compounds disclosed herein.

[0079] G12C mutations in KRAS, HRAS, or NRAS have also been identified in hematological malignancies (e.g., cancers affecting the blood, bone marrow, and / or lymph nodes). Therefore, certain embodiments relate to the administration of the disclosed compounds (e.g., in the form of pharmaceutical compositions) to patients requiring treatment for hematological malignancies. Such malignancies include, but are not limited to, leukemia and lymphoma. For example, the compounds of this disclosure can be used to treat diseases such as 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 another embodiment, the compounds are useful for the treatment of lymphoma, e.g., all subtypes of Hodgkin lymphoma or non-Hodgkin lymphoma. In various embodiments, the compounds are useful in treating plasma cell malignancies such as multiple myeloma, mantle cell lymphoma, and Waldenström macroglobulinemia.

[0080] The determination of whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can be made 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 characterizing the putative KRAS, HRAS, or NRAS mutant protein. The sequences of wild-type human KRAS, HRAS, or NRAS are well known in the art (e.g., accession number NP203524).

[0081] Methods for detecting mutations in KRAS, HRAS, or NRAS nucleotide sequences are well known to those skilled 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 conformational 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 thawing assays, and microarray analysis. In some embodiments, a sample is evaluated for G12C KRAS, HRAS, or NRAS mutations by real-time PCR. In real-time PCR, a fluorescent probe specific to G12C mutations in KRAS, HRAS, or NRAS is used. If a mutation is present, the probe binds and fluorescence is detected. In some embodiments, G12C mutations in KRAS, HRAS, or NRAS are identified using direct sequencing of a specific region (e.g., exon 2 and / or exon 3) within the KRAS, HRAS, or NRAS gene. This method identifies all possible mutations in the sequenced region.

[0082] Methods for detecting mutations in KRAS, HRAS, or NRAS proteins are well known to those skilled in the art. These methods include, but are not limited to, the detection of KRAS, HRAS, or NRAS variants using mutant protein-specific binders (e.g., antibodies), protein electrophoresis and Western blotting, and direct peptide sequencing.

[0083] Various samples can be used in a method for determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation. In some embodiments, the sample is taken from a subject having 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.

[0084] This disclosure also relates to a method for treating a mammalian hyperproliferative disorder, comprising administering a therapeutically effective dose of a compound disclosed herein or a pharmaceutically acceptable salt thereof to the mammal. In some embodiments, the method is used for acute myeloid leukemia, adolescent cancer, pediatric adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical malformation, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, atypical malformation, germ cell tumor, primary lymphoma, cervical cancer, pediatric cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and chronic myeloproliferative disorders. Colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), germ cell tumor, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer, larynx Cancer, lip and oral cancer, liver cancer, carcinoma in situ (LCIS), lung cancer, lymphoma, recurrent metastatic cervical squamous cell carcinoma of unknown primary origin, midline cancer, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma / plasmacytic neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cancer, oropharyngeal cancer, ovarian cancer, This relates to the treatment of patients suffering from cancers such as pancreatic cancer, papilloma, paraganglioma, sinus and nasal cavity cancer, parathyroid cancer, 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, gastric (stomach) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic neoplasm, abnormal cancers in children, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer.In some embodiments, the method relates to the treatment of noncancerous hyperproliferative disorders such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate problems (e.g., benign prostatic hyperplasia (BPH)).

[0085] In some embodiments, the treatment method relates to the treatment of lung cancer, and the method comprises administering an effective amount of any of the above compounds (or pharmaceutical compositions containing them) to a subject in need. 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, adenocarcinoma, carcinoid tumors, and undifferentiated carcinomas.

[0086] The Disclosure further provides a method for modulating the activity of G12C mutant KRAS, HRAS, or NRAS proteins by contacting the protein with an effective amount of the compound of the Disclosure. Modulation may involve inhibiting or activating the protein's activity. In some embodiments, the Disclosure provides a method for inhibiting protein activity by contacting a G12C mutant KRAS, HRAS, or NRAS protein with an effective amount of the compound of the Disclosure in solution. In some embodiments, the Disclosure provides a method for inhibiting the activity of a G12C mutant KRAS, HRAS, or NRAS protein by contacting a cell, tissue, or organ expressing the protein of interest. In some embodiments, the Disclosure provides a method for inhibiting the activity of a protein in a subject, including (but not limited to) rodents and mammals (e.g., humans), by administering an effective amount of the compound of the Disclosure to the subject. In some embodiments, the percentage of modification is greater than 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%.

[0087] In some embodiments, the Disclosure provides a method for inhibiting the G12C activity of KRAS, HRAS, or NRAS in cells by contacting the cells with a compound of the Disclosure in an amount sufficient to inhibit the G12C activity of KRAS, HRAS, or NRAS in the cells. In some embodiments, the Disclosure provides a method for inhibiting the G12C activity of KRAS, HRAS, or NRAS in tissues by contacting the tissue with a compound of the Disclosure in an amount sufficient to inhibit the G12C activity of KRAS, HRAS, or NRAS in the tissue. In some embodiments, the Disclosure provides a method for inhibiting the G12C activity of KRAS, HRAS, or NRAS inside an organism by contacting the organism with a compound of the Disclosure in an amount sufficient to inhibit the G12C activity of KRAS, HRAS, or NRAS inside the organism. In some embodiments, the Disclosure provides a method for inhibiting the G12C activity of KRAS, HRAS, or NRAS within an animal by contacting the animal with a compound of the Disclosure in an amount sufficient to inhibit the G12C activity of KRAS, HRAS, or NRAS within the animal. In some embodiments, the Disclosure provides a method for inhibiting the G12C activity of KRAS, HRAS, or NRAS within a mammal by contacting the mammal with a compound of the Disclosure in an amount sufficient to inhibit the G12C activity of KRAS, HRAS, or NRAS within the mammal. In some embodiments, the Disclosure provides a method for inhibiting the G12C activity of KRAS, HRAS, or NRAS within a human by contacting the human with a compound of the Disclosure in an amount sufficient to inhibit the G12C activity of KRAS, HRAS, or NRAS within the human. The Disclosure provides a method for treating a disease mediated by the G12C activity of KRAS, HRAS, or NRAS, and a method for treating a subject in need of such treatment.

[0088] Combination therapy The Disclosure also provides methods of combination therapy in which agents known to modulate other pathways or other components of the same pathway, or sets of overlapping target enzymes, are used in combination with the compounds of the Disclosure or pharmaceutically acceptable salts thereof. In one embodiment, such therapy includes, but is not limited to, combinations of one or more compounds of the Disclosure with chemotherapeutic agents, therapeutic antibodies, and radiotherapy to provide a synergistic or additive therapeutic effect.

[0089] Many chemotherapeutic agents are currently well known in the art and can be used in combination with the compounds of this disclosure. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples include chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), Venclexta® (venetoclax), and Adriamycin® (doxorubicin), as well as hosts for chemotherapeutic agents. Non-exclusive examples of chemotherapy drugs include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan®); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carbocon, metsuredopa and uredopa; ethyleneimines and methylamelamamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethyloromelamamine; nitrogen mustards such as chlorambucil, chlornafadin, chlorocyclophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobembicin, fenestrine, prednimustine, trophosphamide and uracil mustard; and nitrosoureas such as carmustine, chlorozotosine, fotemustine, lomustine, nimustine and ranimustine.Acrasinomycin, Actinomycin, Ausramycin (Anthramycin), Azaserin, Bleomycin, Kakutinomycin, Calicheamycin, Carabicin, Carminomycin, Cardinophilin, Casodex (trademark), Chromomycin, Dactinomycin, Daunorubicin, Detrubicin, 6-Diazo-5-Oxo-L-Norleucine, Doxorubicin, Epirubicin, Esolubicin, Idarubicin, Marcelomycin, Mitomycin, Mycophenolic acid, Nogaramycin, Olibomycin, Peplomycin, Potrophirom Antibiotics such as Icin, puromycin, keramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, and zolubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, and trimethrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, and doxyl Androgens such as cyfluridine, enocitabine, floxlysine, carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; antiadrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as floric acid; acegraton; aldofhomphamide glycoside; aminolevulinic acid; amsacrin; bestrabusil; bisantren; edatraxate; defofamine; demecolsin; diaziquan; elfomitin; eriptinium acetate; etogluside; gallium nitrate; hydroxy Urea; Lentinan; Ronidamin; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Fenamet; Pirarubicin; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK; Lazoxane; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadiquan; 2,2',2''-Trichlorotriethylamine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacitosine; Arabinoside ("Ara-C"); Cyclophosphamide; Thiotepa;This includes taxanes, such as paclitaxel and docetaxel; retinoic acid; esperamycin; capecitabine; and any pharmaceutically acceptable salts, acids, or derivatives of the above.

[0090] Appropriate chemotherapy cell conditioners include, for example, anti-estrogens including tamoxifen (Nolvadex®), raloxifene, aromatase inhibitor 4(5)imidazole, 4-hydroxytamoxifen, trioxyfen, keoxyfen, LY117018, onapristone, and toremifene (Fareston); anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; and This also includes anti-hormone agents that modulate or inhibit the hormonal effects on tumors, such as mucitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000; and difluoromethylornithine (DMFO).

[0091] If desired, the compounds or pharmaceutical compositions disclosed herein include Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE, avagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alfa-lazine, arbo Sidibu, 3-aminopyridine-2-carboxyaldehyde thiosemicarbazone, amonafide, anthracendione, anti-CD22 immunotoxin, antitumor drug, antitumor-forming herb, apadicone, atiprimod, azathioprine, bendamustine, BIBW2992, bilicodal, brostaricin, bryostatin, butionine sulfoximine, CBV (chemotherapy), kallikrin, cell cycle nonspecific antitumor agent, dichloroacetic acid, discodermorid, erusamitolu Syn, Enocitabine, Epothilon, Eribulin, Everolimus, Exatecan, Exislind, Ferginol, Forodesine, Phosfestrol, ICE chemotherapy regimen, IT-101, Imexone, Imiquimod, Indocarbazole, Ilofluben, Lanikidal, Lalotaxel, Lulutotecan, Maphosfamide, Mitozolomide, Nafoxidin, Nedaplatin, Olaparib, Ortaxel, PAC-1, Pawpaw, Pixantrone, Protea It can be used in combination with commonly prescribed anticancer drugs such as chromosome inhibitors, lebecamycin, reximod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swinesonin, talaporfin, talikidal, tegafur-uracil, temodal, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacin, uramustine, bajimezan, vinflunin, ZD6126, or zoskidal.

[0092] This disclosure further relates to a method of using a compound or pharmaceutical composition provided herein in combination with radiotherapy to inhibit abnormal cell proliferation in mammals or to treat hyperproliferative disorders. Methods for administering radiotherapy are well known in the art and can be used in the combination therapy described herein. The administration of the compounds of this disclosure in this combination therapy can be determined as described herein.

[0093] Radiotherapy can be administered by one of several methods, or a combination thereof, including external beam therapy, internal beam therapy, brachytherapy, stereotactic radiosurgery, total body radiotherapy, radiotherapy, and permanent or transient intratissue brachytherapy. In this specification, the term “brachytherapy” refers to radiotherapy delivered by spatially limited radioactive material inserted into or near a tumor or other proliferative tissue disease site in the body. This term is intended to include, but is not limited to, exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and Lu radioisotopes). Suitable radioactive sources for use as cell conditioners in this disclosure include both solid and liquid sources. As a non-limiting example, the radiation source may 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 rays, or other therapeutic rays. The radioactive material may also be a fluid produced from any solution of a radionuclide, for example, a solution of I-125 or I-131, or the radioactive fluid may be produced using a suitable fluid slurry containing fine particles of solid radionuclides such as Au-198, Y-90. Furthermore, the radionuclide may be embodied in the form of a gel or radioactive microspheres.

[0094] The compounds or pharmaceutical compositions of this disclosure may be used in combination with a certain amount of one or more substances selected from anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors.

[0095] Anti-angiogenic agents such as MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors can be used in combination with the compounds of this disclosure and the pharmaceutical compositions described herein. Examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include arecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are found in International Publication No. 96 / 33172, International Publication No. 96 / 27583, European Patent Application Publication No. 0818442, European Patent Application Publication No. 1004578, International Publication No. 98 / 07697, International Publication No. 98 / 03516, International Publication No. 98 / 34918, International Publication No. 98 / 34915, International Publication No. 98 / 33768, International Publication No. 98 / 30566, European Patent Application Publication No. 0606046, and European Patent Application Publication No. 0931788. These are described in the specifications, International Publication No. 90 / 05719, International Publication No. 99 / 52910, International Publication No. 99 / 52889, International Publication No. 99 / 29667, International Publication No. 1999 / 007675, European Patent Application Publication No. 1786785, European Patent Application Publication No. 1181017, U.S. Patent Application Publication No. 2009 / 0012085, U.S. Patent No. 5863949, U.S. Patent No. 5861510, and European Patent Application Publication No. 0780386, all of which are incorporated herein by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity to inhibit MMP-1. More preferably, it selectively inhibits MMP-2 and / or AMP-9 compared to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP7, MMP8, MMP-10, MMP-11, MMP-12, and MMP-13).Some specific examples of MMP inhibitors useful in this disclosure are AG-3340, RO32-3555, and RS13-0830.

[0096] Furthermore, this compound is also known as acemannan, acralbicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amiphostin, aminolevulinic acid, amrubicin, amsacrin, anagrelide, anastrozole, ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM002 (Novelos), bexarotene, bicalutamide, proxuridine, capecitabine, cermoloukin, cetrorelix, cladribine, clotrimazole, cytarabine ocphosphonate, DA3030 (Dong-A), daciritumab, and Nileukin difutox, Deslorerin, Dexrazoxane, Dilazep, Docetaxel, Docosanol, Doxelcalciferol, Doxifluridine, Doxorubicin, Bromocriptine, Carmustine, Cytarabine, Fluorouracil, HIT Diclofenac, Interferon Alpha, Daunorubicin, Doxorubicin, Tretinoin, Edelfosine, Edrecolomab, Eflornithine, Emitefur, Epirubicin, Epoetin Beta, Etoposide Phosphate, Exemestane, Exislind, Fadrozol, Filgrastim, Finas Teride, fludarabine phosphate, formestan, fotemustine, gallium nitrate, gemcitabine, gemtuzumab ozogamicin, gimeracil / oteracil / tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronate, idarubicin, (imiquimod, interferon alpha, interferon alpha, natural type, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon Alpha-N1, Interferon Alpha-n3, Interferon Alphacon-1, Interferon Alpha, Natural type, Interferon Beta, Interferon Beta-1a, Interferon Beta-1b, Interferon Gamma, Natural type Interferon Gamma-1a, Interferon Gamma-1b, Interleukin-1 Beta, Iobenguan, Irinotecan, Ilsogladine, Lanreotide, LC9018 (Yakult), Leflunomide, Lenograstim, Lentinan sulfate, Letrozole, Leukocyte alpha interferon,Leuprorelin, Levamysol + Fluorouracil, Rialozol, Lovaplatin, Ronidamin, Lovastatin, Masoprocol, Melalsoprole, Metoclopramide, Mifepristone, Miltefosine, Millimostim, Mismatch Double-Stranded RNA, Mitoguazone, Mitractol, Mitoxantrone, Morglamostim, Nafarelin, Naloxone + Pentazocine, Naltograstim, Nedaplatin, Niltamide, Noscapine, Novel Erythrocyte Production Stimulating Protein Preparation, NSC631570 Octreotide, Oprelbequin, Osateron, Oxaliplatin Paclitaxel, Pamidronic acid, PegAsparagase, PegInterferon Alpha-2b, Pentosan sodium polysulfate, Pentostatin, Picibanil, Pirarubicin, Rabbit anti-thymocyte polyclonal antibody, Polyethylene glycol interferon Alpha-2a, Porfimer sodium, Raloxifene, Larcitrexed, Rasburiembodiment, Rhenium etidronate Re186, RII retinamide, Rituximab, Romultide, Samarium (153Sm) lexidronam, Salg Ramostim, schizophyllan, sobuzoxane, sonelmin, strontium-89 chloride, suramin, tasonelmin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecoxide, thalidomide, thymalfacin, thyrotropin alpha, topotecan, toremifene, tositumomab-iodine-131, trastuzumab, treosulfan, tretinoin, trilostane, trimethrexate, triptorelin, tumor necrosis factor alpha, natural type, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysis vaccine, ba Lurubicin, verteporfin, vinorelbine, VIRULIZIN, dinostatin stimulamer, or zoledronic acid; Abarelix; AE941 (Aeterna), ambamustin, antisense oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diaziquan, EL532 (Elan), EM800 (Endorecherche), enyluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant,Galocitabine, Gastrin 17 immune antigen, HLA-B7 gene therapy drug (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin 2, iproxyfen, LDI200 (Milkhaus), religistim, lintuzumab, CA125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and FcMAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), LYM-1-iodine 131 MAb (Techniclone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), Marimast, Menogalil, Mitsumomab, Motexafingadolinium, MX6 (Galderma), Nelarabine, Noratexed, P30 protein, Pegvisomant, Pemetrexed, Porphyromycin, Prinomast, RL0903 (Shire), Rubitecan, Satraplatin, Sodium phenylacetate, Sparfosic acid, SRL172 (SRPharma), SU5416 (SUGEN, now Pfizer.Inc.), TA077 (Tanabe), Tetrathiomolybdate, Talibrasticin, Thrombopoietin, Tin-ethylethiopurine, Tirapazamine, Cancer vaccine (Biomira), Melanoma vaccine (New York University), Melanoma vaccine (Sloan Kettering Institute), Melanoma tumor degeneration vaccine (New York Medical College), Viral melanoma cell lysate vaccine (Royal Newcastle It may be used in combination therapy with other antitumor drugs, such as Valspodar (or other antitumor drugs).

[0097] The compounds of this disclosure may be used in combination with VEGFR inhibitors. In combination therapy, other compounds described in the following patents and patent applications may be used: U.S. Patent No. 6,258,812, U.S. Patent Application Publication No. 2003 / 0105091, International Publication No. 01 / 37820, U.S. Patent No. 6,235,764, International Publication No. 01 / 32651, U.S. Patent No. 6,630,500, U.S. Patent No. 6,515,004, U.S. Patent No. 6,713,485, U.S. Patent No. 5,521,184, U.S. Patent No. 5,770,599, U.S. Patent No. 5,747,498, International Publication No. 02 / 68406 Brochures, International Publication No. 02 / 66470, International Publication No. 02 / 55501, International Publication No. 04 / 05279, International Publication No. 04 / 07481, International Publication No. 04 / 07458, International Publication No. 04 / 09784, International Publication No. 02 / 59110, International Publication No. 99 / 45009, International Publication No. 00 / 59509, International Publication No. 99 / 61422, U.S. Patent No. 5,990,141, International Publication No. 00 / 12089, and International Publication No. 00 / 02871.

[0098] In some embodiments, the combination agent comprises the composition of the present disclosure in combination with at least one anti-angiogenic agent. The agent includes, but is not limited to, chemical compositions, antibodies, antigen-binding domains, radionuclides, and combinations and conjugates thereof, prepared by synthesis in vitro. The agent may be an agonist, antagonist, allosteric modulator, toxin, or, more generally, it may act to inhibit or stimulate its target (e.g., activation or inhibition of receptors or enzymes) thereby promoting cell death or inhibiting cell proliferation.

[0099] Exemplary anti-angiogenic drugs include ERBITUX® (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to kinase domain receptors), anti-VEGF drugs such as AVASTIN® or VEGF-TRAP® (e.g., antibodies or antigen-binding regions that specifically bind to VEGF or soluble VEGF receptors or their ligand-binding regions), anti-VEGF receptor drugs (e.g., antibodies or antigen-binding regions that specifically bind thereto), Vectibix (panitumumab), IRESSA® (gefitinib), TARCEVA® (erlotinib), anti-Ang1 and anti-Ang2 drugs (e.g., antibodies or antigen-binding regions that specifically bind thereto or their receptors, e.g., Tie2 / Tek), and EGFR inhibitors such as anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto). The pharmaceutical compositions of this disclosure may also include one or more agents (e.g., an antibody, an antigen-binding domain, or a soluble receptor) such as an antagonist of hepatocyte growth factor (HGF, also known as a scattering factor) that specifically binds and inhibits the activity of the growth factor, and an antibody or antigen-binding domain that specifically binds to the receptor "c-met".

[0100] Other anti-angiogenic drugs include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., U.S. Patent Application Publication No. 2003 / 0162712; U.S. Patent No. 6,413,932), anti-TWEAK agents (e.g., antibodies or antigen-binding domains that specifically bind; or soluble TWEAK receptor antagonists; see Wiley, U.S. Patent No. 6,727,225), ADAM disintegrin domains that antagonize the binding of integrins to their ligands (Fanslow et al., U.S. Patent Application Publication No. 2002 / 0042368), and anti-eph that specifically bind. These include receptor antibodies and / or anti-ephrin antibodies or antigen-binding regions (U.S. Patent Nos. 5,981,245; Nos. 5,728,813; Nos. 5,969,110; Nos. 6,596,852; Nos. 6,232,447; Nos. 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 to PDGF-BB ligands, and PDGFR kinase inhibitors (e.g., antibodies) and antibodies or antigen-binding regions that specifically bind to them).

[0101] Further anti-angiogenic / antinomatous agents include: SD-7784 (Pfizer, USA); Silendide (Merck KGaA, Germany, European Patent No. 770622); Pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, U.S. Patent No. 5712291); Ilomast (Arriva, USA, U.S. Patent No. 5892112); Emaxanib (Pfizer, USA, U.S. Patent No. 5792783); Batalanib (Novartis, Switzerland); 2-Methoxyestradiol (EntreMed, now CASI Pharmaceuticals, USA); TLC ELL-12 (Elan, Ireland); Anecol Acetate Tab (Alcon, USA); Alpha-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); Anti-Vn Mab (Crucell, Netherlands); DAC: Anti-angiogenic drug (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, European Patent No. 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, (EntreMed, USA); Angiogenesis Inhibitor, (Tripep, Sweden); Maspin, (Sosei Group Corporation, Japan); 2-Methoxyestradiol, (Oncology Sciences Corporation, USA); ER-68203-00, (IVAX, USA); Benefin, (Lane Labs, USA);Tz-93 (Tsumura Co., Ltd., Japan); TAN-1120 (Takeda Pharmaceutical Company Limited, Japan); FR-111142 (Fujisawa Pharmaceutical Co., Ltd., Japan, Japanese Patent Publication No. 02-233610); Platelet Factor IV (RepliGen, USA, European Patent No. 407122); Vascular Endothelial Growth Factor Antagonist (Borean, Denmark); Bevacizumab (pINN), (Genentech, USA); Angiogenesis Inhibitor (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); MAb, Alpha 5 Beta 3 Integrin, Second Generation (Applied Molecular Evolution, USA and MedImmune, USA); Gene Therapy, Retinopathy, (Oxford BioMedica, 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-inducing anti-angiogenic agent, (XOMA, USA); PI 88, (Progen, Australia); Silentide (pINN), (Merck KGaA, Germany; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); Cetuximab (INN), (Aventis, France); AVE 8062, (Ajinomoto Co., Inc., Japan); AS 1404, (Cancer Research Laboratory, New Zealand); SG 292, (Telios, USA); Endostatin, (Boston Children's Hospital, USA); ATN 161, (Attenuon, USA); Angiostatin, (Boston Children's Hospital, USA); 2-Methoxyestradiol, (Boston Children's Hospital, USA);ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); 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); xanthrizole, (Yonsei University, South Korea); vaccine, gene-based, VEGF-2, (Scripps Clinic and Research Foundation, USA); SPV5.2, (Supratek, Canada); SDX 103, (University of California at San Diego, USA); PX 478, (ProlX, USA); METASTATIN, (EntreMed, now CASI Pharmaceuticals, USA); Troponin I, (Harvard University, USA); SU 6668, (SUGEN, now Pfizer, Inc., USA); OXI 4503, (OXiGENE, USA); o-Guanidine, (Dimensional Pharmaceuticals, USA); Motupolamine C, (British Columbia University, Canada); CDP 791, (Celltech Group, UK); Atiprimod (pINN), (GlaxoSmithKline, UK); E 7820, Eisai Co., Ltd., Japan); CYC 381, (Harvard University, USA); AE 941, (Aeterna, Canada); Vaccine, Angiogenic Drug, (EntreMed, now CASI Pharmaceuticals, USA; Urokinase-type plasminogen activator inhibitor, (Dendreon, USA); Oglufanide (pINN), (Melmotte, USA); HIF-1 alpha inhibitor, (Xenova, UK); CEP 5214, (Cephalon, USA);BAY RES 2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom, USA); KR31372, (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 Company, Limited, Japan); Drug delivery system, intraocular drug, 2-methoxyestradiol, (EntreMed, USA); Anginex, (Maastricht University, Netherlands, and Minnesota University, USA); ABT 510, (Abbott, USA); AAL 993, (Novartis, Switzerland); VEGI, (ProteomTech, USA); Tumor Necrosis Factor-Alpha Inhibitor, (National Institute on Aging, USA); SU 11248, (Pfizer, USA and SUGEN USA); ABT 518, (Abbott, USA); YH16, (Yantai Rongchang, China); S-3APG, (Boston Children's Hospital, USA and EntreMed, USA); MAb, KDR, (ImClone Systems, USA); MAb, Alpha 5 Beta 1, (Protein Design, USA); KDR Kinase Inhibitor, (Celltech Group, UK and Johnson & Johnson, USA); GFB 116, (South Florida University, USA and Yale University, USA); CS 706, (Sankyo Co., Ltd., Japan); Combretastatin A4 Prodrug, (Arizona State University, USA; Chondroitinase AC, (IBEX, Canada); BAY RES 2690, (Bayer, Germany);AGM 1470 (Harvard University, USA, Takeda Pharmaceutical Company Limited, Japan, and TAP, USA); AG 13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); MAb, Vascular Endothelial Growth Factor (Xenova, UK); Irsoglandin (INN), (Nippon Shinyaku Co., Ltd., Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); Squalamine (pINN), (Genaera, USA); RPI 4610 (Sirna, USA); Cancer Therapy (Marinova, Australia); Heparanase Inhibitor (InSight, Israel); KL 3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho Pharmaceutical Co., Ltd., Japan); VEGF receptor modulator (Pharmacopeia, USA); VE-cadherin-2 antagonist (ImClone Systems, USA); Vasostatin (National Institutes of Health, USA); Vaccine, Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura & Co., Ltd., Japan); Tumustatin (Beth Israel Hospital, USA); Sclerolytic soluble FLT 1 (vascular endothelial growth factor receptor 1), (Merck & Co, USA); Tie-2 ligand, (Regeneron, USA); and thrombospondin 1 inhibitor, (Allegheny Health, Education and Research Foundation, USA).

[0102] Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil®), bafilomycin A1, 5-amino-4 imidazole carboxamidriboside (AICAR), okadaic acid, autophagy-suppressing algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and agents that increase cAMP levels, such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Furthermore, antisense or siRNAs that inhibit the expression of proteins including (but not limited to) ATG5 (involved in autophagy) may also be used.

[0103] Further pharmaceutically active compounds / agents that can be used in the treatment of cancer and in combination with one or more of the compounds of this disclosure include epoetin alfa, darbepoetin alfa, panitumumab, pegfilgrastim, palifermin, filgrastim, denosumab, ancestim, AMG102, AMG176, AMG386, AMG479, AMG655, AMG745, AMG951, AMG706, or pharmaceutically acceptable salts thereof.

[0104] In certain embodiments, the compositions provided herein are administered together with chemotherapeutic agents. Suitable chemotherapeutic agents include vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), paclitaxel, epidipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mitramycin), mitomycin, and enzymes (e.g., enzymes capable of systemically metabolizing L-asparagine and synthesizing asparagine itself). L-asparaginase (which removes cells that do not possess), antiplatelet agents, antiproliferative / antimitotic alkylating agents such as nitrogen mustard (e.g., mechloretamine, cyclophosphamide and its analogs, melphalan and chlorambucil), ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., sericiclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638 and SCH727965), alkylsul Antiproliferative / antimitotic antimetabolites such as fonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and its analogues, as well as streptozocin), trazene-dacarbadinine (DTIC), folic acid analogues (e.g., methotrexate), pyrimidine analogues (e.g., fluorouracil, phloxuridine, and cytarabine), purine analogues and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, 2-chlorodeoxyadenosine), and aromatase inhibitors (e.g., anastrozole, exocrine). Mestane, letrozole) and platinum-coordinate complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apisidan, suberoylanilide hydroxamic acid, vorinostat, LBH589, romidepsin, ACY-1215, and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridafololimus, and sirolimus), KSP (Eg5) inhibitors (e.g.,Examples include Array520), DNA binders (e.g., Zalypsis), PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), multikinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen), and leutinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide, and triptorelin). Hormone agonists, BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN163L), 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 KOS95) 3) P13K / Akt inhibitors (e.g., Perifosin), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., Enzastaurin), FTIs (e.g., Zanestra®), 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 ( For example, natural products such as ARRY-382, JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), and BCL-2 antagonists are included. Other chemotherapeutic agents may include mechloretamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or any analogues or induced variants of the above.

[0105] The compounds disclosed herein may also be used in combination with radiotherapy, hormone therapy, surgery, and immunotherapy, which are well known to those skilled in the art.

[0106] In certain embodiments, the pharmaceutical compositions provided herein are administered together with a steroid. Suitable steroids include 21-acetoxypregnenolone, alclomethasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, crocortol, cloprednol, corticosterone, cortisone, cortibazole, deflazacort, desonide, dexoxymethasone, dexamethasone, diflorason, diflucortol, difluprednate, enoxolone, fluazacort, fluchloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, flupredniden acetate, fluprednisolone, and flulane. This includes, but is not limited to, drenolide, fluticasone propionate, formocortal, halcinonide, halobetazole propionate, halomethasone, hydrocortisone, loteprednol etabonate, mazipredone, medrisone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, thixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and / or salts and derivatives thereof. In certain embodiments, the compounds of this disclosure may also be used in combination with further pharmaceutically active agents for treating nausea. Examples of medications that can be used to treat nausea include dronabinol; granisetron; metoclopramide; ondansetron; and prochlorperazine; or their pharmaceutically acceptable salts.

[0107] The compounds of this disclosure may also be used in combination with further pharmaceutically active compounds that disrupt or inhibit the RAS-RAF-ERK or PI3K-AKT-TOR signaling pathway. Other such combinations include PD-1 and PD-L1 antagonists. The compounds or pharmaceutical compositions of this disclosure may also be used in combination with any 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 immunotherapeutic agents (e.g., 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 BiTE).

[0108] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux), panitumumab (Vectibix), zaltumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and more recently, lapatinib (TykerB). For example, see Yan L, et al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, Bio Techniques 2005;39(4):565-8, and Paez JG, et al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004;304(5676):1497-500.

[0109] Non-limiting examples of small molecule EGFR inhibitors include any EGFR inhibitors described in the following patent publications, as well as all pharmaceutically acceptable salts and solvates thereof: European Patent Application Publication No. 520722, published December 30, 1992; European Patent Application Publication No. 566226, published October 20, 1993; International Publication No. 96 / 33980, published October 31, 1996; U.S. Patent No. 5,747,498, issued May 5, 1998; and October 3, 1996. International Publication No. 96 / 30347 published on [date]; Specification of European Patent Application Publication No. 787772 published on August 6, 1997; International Publication No. 97 / 30034 published on August 21, 1997; International Publication No. 97 / 30044 published on August 21, 1997; International Publication No. 97 / 38994 published on October 23, 1997; International Publication No. 97 / 49688 published on December 31, 1997; European Patent Application Publication No. 837063 published on April 22, 1998 Specifications; International Publication No. 98 / 02434 published on January 22, 1998; International Publication No. 97 / 38983 published on October 23, 1997; International Publication No. 95 / 19774 published on July 27, 1995; International Publication No. 95 / 19970 published on July 27, 1995; International Publication No. 97 / 13771 published on April 17, 1997; International Publication No. 98 / 02437 published on January 22, 1998; published on January 22, 1998 International Publication No. 98 / 02438; International Publication No. 97 / 32881, published on September 12, 1997; German Patent Application Publication No. 19629652, published on January 29, 1998; International Publication No. 98 / 33798, published on August 6, 1998; International Publication No. 97 / 32880, published on September 12, 1997; International Publication No. 97 / 32880, published on September 12, 1997; European Patent Application Publication No. 682027, published on November 15, 1995;International publication pamphlet No. 97 / 02266, published on January 23, 1997; International publication pamphlet No. 97 / 27199, published on July 31, 1997; International publication pamphlet No. 98 / 07726, published on February 26, 1998; International publication pamphlet No. 97 / 34895, published on September 25, 1997; International publication pamphlet No. 96 / 31510, published on October 10, 1996; International publication pamphlet No. 98 / 14449, published on April 9, 1998; International publication pamphlet No. 98 / 14450, published on April 9, 1998; International publication pamphlet No. 98 / 14451, published on April 9, 1998; International publication pamphlet No. 95 / 09847, published on April 13, 1995 T; International Publication No. 97 / 19065, published on 29 May 1997; International Publication No. 98 / 17662, published on 30 April 1998; U.S. Patent No. 5,789,427, issued on 4 August 1998; U.S. Patent No. 5,650,415, issued on 22 July 1997; U.S. Patent No. 5,656,643, issued on 12 August 1997; International Publication No. 99 / 35146, published on 15 July 1999; International Publication No. 99 / 35132, published on 15 July 1999; International Publication No. 99 / 07701, published on 18 February 1999; and International Publication No. 92 / 20642, published on 26 November 1992. Further non-exclusive examples of small molecule EGFR inhibitors include all EGFR inhibitors listed in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12):1599-1625.

[0110] Antibody-based EGFR inhibitors include all anti-EGFR antibodies or antibody fragments that can partially or completely block EGFR activation by their natural ligands. Non-exclusive examples of antibody-based EGFR inhibitors are described in 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, SM, et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang, X., et al., 1999, Cancer Res. 59:1236-1243. Therefore, EGFR inhibitors may be monoclonal antibodies such as Mab E7.6.3 (Yang, 1999, cited above), or Mab C225 (ATCC accession number HB-8508), or antibodies or antibody fragments having binding specificity thereto.

[0111] KRAS of this disclosure G12CThe inhibitor can be used in combination with a MEK inhibitor. Specific MEK inhibitors that can be used in combination with the inhibitors described herein include PD-325901, trametinib, pimacertib, MEK162 [also known as binimetinib], TAK-733, GDC-0973, and AZD8330. A specific MEK inhibitor that can be used in combination with a KRASG12C inhibitor as described herein is trametinib (trade name: Mekinist®, marketed by 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 AMG1009089, 1009089, or PD-325901. Another specific MEK inhibitor that can be used in combination with the inhibitors described herein is cobimetinib. MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, and ARRY-438162.

[0112] PI3K inhibitors include woltmannin, 17-hydroxywoltmannin analog, 4-[2-(1H-indazole-4-yl)-6-[[4-(methylsulfonyl)piperazine-1-yl]methyl]thieno[3,2-d]pyrimidine-4-yl]morpholine (also known as GDC 0941, described in International Publications 09 / 036,082 and 09 / 055,730), 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinoline-3-yl)-2,3-dihydroimidazo[4,5-c]quinoline-1-yl]phenyl]propionitrile (also known as BEZ235 or NVP-BEZ235, described in International Publication 06 / 122806). (As listed in the fret), (S)-1-(4-((2-(2-aminopyrimidine-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidine-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (as listed in International Publication No. 2008 / 070740), LY294002(2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, Axon PI103 hydrochloride (available from Medchem), PI103 hydrochloride (3-[4-(4-morpholinylpyrido-[3',2':4,5]fl[3,2-d]pyrimidine-2-yl]phenol hydrochloride, available from Axon Medchem), PIK 75 (N'-[(1E)-(6-bromoimidazo[1,2-a]pyridine-3-yl)methylene]-N,2-dimethyl-5-nitrobenzene sulfon-hydrazide hydrochloride, available from Axon Medchem), PIK90 (N-(7,8-dimethoxy-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl)-nicotinamide, Axon (Available from Medchem), GDC-0941 bismesylate (2-(1H-indazole-4-yl)-6-(4-methanesulfonyl-piperazine-1-ylmethyl)-4-morpholine-4-yl-thieno[3,This includes, but is not limited to, 2-d]pyrimidine (available from Axon Medchem), AS-252424 (5-[1-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidin-2,4-dione (available from Axon Medchem), and TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidine-4-one (available from Axon Medchem), XL-765, and XL-147. Other PI3K inhibitors include demethoxypyridine, perifosin, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0113] Examples of AKT inhibitors include Akt-1-1 (inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and Ak2) (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., International Publication No. 05 / 011700); indole-3-carbinol and its derivatives (e.g., U.S. Patent No. 6,656,963; Sarkar and Li (2004) J Nutr. 134 (12) Examples include, but are not limited to, Suppl), 3493S-3498S; perifosin (e.g., interfering with the membrane localization of Akt; 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 trisirivine (TCN or API-2 or NCI identifier: NSC154020; Yang et al. (2004) Cancer Res. 64, 4394-9).

[0114] TOR inhibitors include, but are not limited to, AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, and ATP-competitive TORC1 / TORC2 inhibitors (including PI-103, PP242, PP30, and Torin1). Other TOR inhibitors include FKBP12 enhancers; rapamycin and its derivatives (including: CCI-779 (temsirolimus), RAD001 (everolimus); International Publication No. 9409010) and AP23573; rapamycin and its derivatives disclosed in, for example, International Publication No. 98 / 02441 and International Publication No. 01 / 14387, e.g., AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl) rapamycin Syn, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also known as CC1779), 40-epi-(tetrazolyl)-rapamycin (also known as ABT578), 32-deoxorapamycin, 16-pentinyloxy-32(S)-dihydrorapamycin, and other derivatives disclosed in International Publication No. 05005434); U.S. Patent No. 5,258,389, International Publication No. 94 / 090101 Brochures, International Publication No. 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, International Publication No. 93 / 111130, International Publication No. 94 / 02136, International Publication No. 94 / 02485, International Publication No. 95 / 14023 This includes derivatives disclosed in International Publication No. 94 / 02136, International Publication No. 95 / 16691, International Publication No. 96 / 41807, International Publication No. 96 / 41807 and U.S. Patent No. 5,256,790; phosphorus-containing rapamycin derivatives (e.g., International Publication No. 05016252); and 4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional Patent Application No. 60 / 528,340).

[0115] MCl-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the important anti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family. Overexpression of MCL-1 has been closely associated with tumor progression and resistance to targeted therapies, including BCL-2 inhibitors such as ABT-263, as well as to conventional chemotherapy.

[0116] KRAS G12C Inhibitors may also be used in combination with SHP2 inhibitors as described in this disclosure. SHP2 inhibitors that may be used in combination therapies as described in this disclosure include, but are not limited to, SHP099, and RMC-4550 or RMC-4630 manufactured by Revolutions Medicines, Redwood City, CA.

[0117] Proteasome inhibitors include, but are not limited to, Kyprolis® (carfilzomib), Velcade® (bortezomib), and oprozomib.

[0118] Immunotherapy includes, but is not limited to, anti-PD-1 drugs, anti-PD-L-1 drugs, anti-CTLA-4 drugs, anti-LAG1 drugs, and anti-OX40 drugs.

[0119] Monoclonal antibodies include, but are not limited to, Darzalex® (daratumumab), Herceptin® (trastuzumab), Avastin® (bevacizumab), Rituxan® (rituximab), Lucentis® (ranibizumab), and Eylea® (aflibercept).

[0120] Immunomodulators (IMiDs) are a class of immunomodulatory drugs (drugs that modulate the immune response) that contain an imide group. The IMiD class includes thalidomide and its analogues (lenalidomide, pomalidomide, and apremilast).

[0121] Anti-PD-1 inhibitors, including but not limited to antibodies, include, but are not limited to, pembrolizumab (Keytruda®), AMG-404, and nivolumab (Opdivo®). Exemplary anti-PD-1 antibodies and their uses are described in Goldberg et al., Blood 110(1):186-192 (2007), Thompson et al., Clin. Cancer Res. 13(6):1757-1761 (2007), and Korman et al., International Application No. PCT / JP2006 / 309606 (International Publication No. 2006 / 121168 A1), each of which is expressly incorporated herein by reference. Contains: Yervoy® (ipilimumab) or tremelimumab (for CTLA-4), galiximab (for B7.1), BMS-936558 (for PD-1), MK-3475 (for PD-1), AMP224 (for B7DC), BMS-936559 (for B7-H1), MPDL3280A (for B7-H1), MEDI-570 (for ICOS), AMG557 (for B7H2), MGA271 (for B7H3), IMP321 (for LAG-3), BMS-663513 (for CD137), PF-05082566 (for CD137), CDX-1127 (for CD27), anti-OX40 (Providence Health Services), huMAbOX40L (to OX40L), atacicept (to TACI), CP-870893 (to CD40), lucatumumab (to CD40), dacetuzumab (to CD40), muromonab-CD3 (to CD3), ipilimumab (to CTLA-4). Immunotherapy also includes genetically modified T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTE).

[0122] GITR agonists include GITR fusion proteins as described in U.S. Patent No. 6,111,090 box.c, European Patent No. 090505B1, U.S. Patent No. 8,586,023, International Publication No. 2010 / 003118 and International Publication No. 2011 / 090754, or, for example, 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. 1866339, International Publication No. 2011 / 028683 and International Publication No. 2013 / 039954. This includes, but is not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as the anti-GITR antibodies described in International Publication No. 2005 / 007190, International Publication No. 2007 / 133822, International Publication No. 2005 / 055808, International Publication No. 99 / 40196, International Publication No. 2001 / 03720, International Publication No. 99 / 20758, International Publication No. 2006 / 083289, International Publication No. 2005 / 115451, U.S. Patent No. 7,618,632, and International Publication No. 2011 / 051726.

[0123] The compounds described herein can be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated. Accordingly, in some embodiments, one or more of the compounds disclosed herein will be administered concurrently with the other agents described above. When used in combination therapy, the compounds described herein are administered concurrently or separately with a second agent. This concurrent administration may include concurrent administration of two agents in the same dosage form, concurrent administration of different dosage forms, and separate administrations. That is, both the compounds described herein and the agents described above can be combined into the same dosage form and administered concurrently. Alternatively, both the compounds disclosed herein and the agents described above can be administered concurrently, where both agents are in separate dosage forms. In another alternative, the compounds disclosed may be administered immediately before or after any of the agents described above. In some embodiments of the separate administration protocol, the compounds disclosed and any of the agents described above are administered with a gap of several minutes, several hours, or several days between them.

[0124] Since one aspect of this disclosure is intended for the treatment of a disease / condition by a combination of pharmaceutically active compounds that can be administered separately, this disclosure further relates to combining separate pharmaceutical compositions into a kit form. The kit comprises two separate pharmaceutical compositions: a compound of this disclosure and a second pharmaceutical compound. The kit includes a container for housing the separate compositions, such as a divided bottle or a divided foil packet. Further examples of containers include syringes, boxes, and bags. In some embodiments, the kit includes instructions for the use of the separate components. The kit form is particularly advantageous when the separate components are administered in different forms of dosing (e.g., orally and parenterally), at different dosing intervals, or when the prescribing healthcare professional desires to set the dosages of the individual components of the combination.

[0125] All patents and other publications listed herein are incorporated herein by reference.

[0126] The steps described below illustrate specific embodiments of this disclosure. These steps are representative and do not limit the scope of the claims in any way.

[0127] Related processes 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]pyrimidine-2(1H)-one are representative examples of the present disclosure and should not be construed as limiting the scope of the present invention.

[0128] The synthesis of compound 9 and related intermediates is described in U.S. Patent Application No. 15 / 984,855, filed on 21 May 2018 (U.S. Patent Application Publication No. 2018 / 0334454, November 22, 2018), claiming priority and benefit of U.S. Provisional Patent Application No. 62 / 509,629, filed on 22 May 2017, both of which 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]pyrimidine-2(1H)-one was prepared by the following method, isolating the final product isomer by chiral chromatography. [ka]

[0129] Another synthesis of compound 9 and related intermediates is described in a U.S. provisional patent application filed on November 16, 2018, which is incorporated herein by reference in its entirety for all purposes. [ka]

[0130] This disclosure includes the following steps, in which the synthesis and use of the boroxine intermediate constitute a novel and progressive step in the production of AMG 510 (compound 9). [ka]

[0131] [Table 4]

[0132] Process 1a [ka]

[0133] [Table 5]

[0134] 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), oxalyl chloride (18.9 kg; 148.9 mol) was added while maintaining the internal temperature at 15-20°C. Additional dichloromethane (33 kg) was added as a rinse, and the reaction mixture was stirred for 2 hours. The reaction mixture was cooled, and then the reaction was stopped with ammonium hydroxide (40.2 L; 595.5 mol) while maintaining the internal temperature at 0 ± 10°C. The resulting slurry was stirred for 90 minutes, and the product was recovered by filtration. The filtered solid was washed with DI water (3 × 87 L) and dried to obtain 2,6-dichloro-5-fluoronicotinamide (compound 1).

[0135] Process 1b [ka]

[0136] [Table 6]

[0137] 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 a temperature of ≤25°C for 75 minutes. The resulting solution was then heated to 40°C ± 3°C and aged for 3 hours. Using vacuum, the solution was distilled to remove dichloromethane until the solution was below the stirrer. Then, dichloromethane (300 kg) was added, and the mixture was cooled to 0 ± 5°C. In a clean, dry reactor (reactor B), 2-isopropyl-4-methylpyridine-3-amine (ANILINE compound 2A) (12.9 kg; 85.9 mol), followed by dichloromethane (102.6 kg), was added. The ANILINE solution was azeotropically dried by vacuum distillation, replacing it with further dichloromethane until the solution was dry (limit ≤0.05%) by KF analysis, while maintaining the internal temperature between 20 and 25°C. The volume of the solution was adjusted to approximately 23 L using dichloromethane. The dried ANILINE solution was then added to reactor A (while maintaining an internal temperature of 0 ± 5°C). The mixture was then heated to 23°C and aged for 1 hour. The solution was filtered into a clean reactor to obtain 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridine-3-yl)carbamoyl)nicotinamide (compound 3) as a solution in DCM, which was used directly in the next step.

[0138] Process 2 [ka]

[0139] [Table 7]

[0140] While maintaining an internal temperature of 20-25°C, a dichloromethane solution of 2,6-dichloro-5-fluoro-N-{[4-methyl-2-(propan-2-yl)pyridine-3-yl]carbamoyl}pyridine-3-carboxamide (UREA (compound 3)) (15 kg; 38.9 mol) was converted to 2-MeTHF by vacuum distillation. The reactor volume was adjusted to 40 L, and then an additional 2-MeTHF (105.4 kg) was added. While maintaining a temperature of 5-10°C, sodium t-butoxide (9.4 kg; 97.8 mol) was added. The contents were warmed to 23°C and stirred for 3 hours. Then, the contents were cooled to 0-5°C, and ammonium chloride (23.0 kg; 430 mol) was added as a solution in 60 L of DI water. The mixture was warmed to 20°C, DI water (15 L) was added, and aging was carried out for a further 30 minutes. Stirring was stopped and the layers were separated. The aqueous layer was removed, and DI 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 pH was measured to be 4-5. The layers were separated, and the aqueous layer was back-extracted using 2-MeTHF (42.2 kg). The two organic layers were combined and washed with 10% citric acid solution (75 kg), followed by a mixture of water (81.7 L) and saturated NaCl (19.8 kg). The organic layers were then washed with saturated sodium bicarbonate (75 kg) to achieve the target pH of the aqueous solution ≥ 7.0 (washing was repeated if necessary). The organic layers were 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 vacuum distilled to a volume of 40 L. The concentrated solution was heated to 55°C, and heptane (10-12 kg) was slowly added until the cloud point was reached. The solution was cooled to 23°C over 2 hours, and then heptane (27.3 kg) was added over 2 hours. The product slurry was aged at 20-25°C for 3 hours, then filtered and washed with a mixture of 2-MeTHF (2.8 kg) and heptane (9 kg). The product was dried using nitrogen and vacuum to obtain the solid 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridine-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (rac-DIONE (compound 4)).

[0141] Process 3 [ka]

[0142] [Table 8]

[0143] Under a nitrogen atmosphere, a stirred suspension of compound 4 (1.0 equivalent) in 2-methylterahydrofuran (7.0 L / kg) was added to a container along with (+)-2,3-dibenzoyl-D-tartaric acid (2.0 equivalents). Although 2-MeTHF is chiral, it is used as a racemic mixture. Different enantiomers of 2-MeTHF are randomly incorporated into the cocrystal. The resulting suspension was heated to 75°C and aged at 75°C until complete dissolution was observed (≤30 minutes). The resulting solution was filtered at 75°C into a second container. To the filtered solution, n-heptane (2.0 L / kg) was added at a rate that maintained the internal temperature above 65°C. The solution was then cooled to 60°C, crystal seeds (0.01 kg / kg) were added, and the mixture was 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 added to the suspension, and the mixture was then aged under a nitrogen atmosphere at 20°C for 4 hours. The suspension was filtered, and the isolated solid was washed twice with (2:1) n-heptane:2-methyltetrahydrofuran (3.0 L / kg). This material was dried under nitrogen and vacuum to obtain the M-dione:DBTA:Me-THF complex (compound 4a).

[0144] Process 4 [ka]

[0145] [Table 9]

[0146] A suspension of disodium hydrogen phosphate (21.1 kg, 2.0 equivalents) in DI water (296.8 L, 6.3 L / kg) was stirred towards container A until dissolution was observed (≥30 minutes). A suspension of M-dione:DBTA:Me-THF complex (composition 4a) [46.9 kg (corrected for M-dione to 25.9 kg, 1.0 equivalent)] in methyl tert-butyl ether (517.8 L, 11.0 L / kg) was stirred towards container B for 15-30 minutes. The solution obtained from container A was added to container B, and the mixture was then stirred for more than 3 hours. Stirring was stopped, and the two-phase mixture was left to stand for more than 30 minutes for separation. The lower aqueous phase was removed, and then back-extracted with methyl tert-butyl ether (77.7 L, 1.7 L / kg). The organic phases were combined in container B and dried over magnesium sulfate (24.8 kg, 0.529 kg / kg). The suspension obtained from container B was stirred for over 3 hours, and then filtered into container C. Methyl tert-butyl ether (46.9 L, 1.0 L / kg) rinse solution was added to container B, and then filtered into container C. The contents of container C were cooled to 10°C, and then distilled under vacuum while slowly heating to 35°C. Distillation was continued until 320-350 kg (6.8-7.5 kg / kg) of methyl tert-butyl ether was recovered. After cooling the contents of container C to 20°C, n-heptane (278.7 L, 5.9 L / kg) was added over 1 hour, and then vacuum distillation was carried out while slowly heating to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane was recovered. After cooling the contents of container C to 20°C, a second dose of n-heptane (278.7 L, 5.9 L / kg) was added over 1 hour, and then vacuum distillation was performed while slowly heating to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg / kg) of the methyl tert-butyl ether and n-heptane mixture was recovered. After cooling the contents of container C to 20°C, a third dose of n-heptane (195.9 L, 4.2 L / kg) was added over 1 hour, and then a sample was taken for solvent composition analysis by GC. The suspension in container C was stirred for more than 1 hour. The suspension was filtered and then washed with the n-heptane rinse solution (68.6 L, 1.5 L / kg) from container C.The isolated solid was dried at 50°C, and the sample was submitted for storage suitability. Compound 5M of 7-chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridine-3-yl]pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (M-DIONE) was obtained.

[0147] In the first-generation process highlighted above, scaling was feasible with 200+ kg of rac-dione starting material (compound 4). In this process, adding a thermodynamically stable rac-dione crystalline form (which exhibits low solubility) would lead to batch failure. Based on our subsequent research, we found that increasing the DBTA equivalent and lowering the seed addition temperature by adjusting the heptane addition schedule improved the robustness of the process. The improved method is resistant to the presence of thermodynamically stable rac-dione crystalline forms and facilitates successful atropisomer separation. Subsequent batches will incorporate the improved process for large-scale production.

[0148] Process 5 [ka]

[0149] [Table 10]

[0150] M-dione (compound 5M, 1.0 equivalent) and toluene-1 (10.0 L / kg) were added to container A. The resulting solution was dried by azeotropic distillation under vacuum at 45°C until 5.0 L / kg of solvent was removed. The contents of container A were then cooled to 20°C.

[0151] To container C, toluene-3 (4.5 L / kg), phosphoryl chloride (1.5 equivalents), and N,N-diisopropylethylamine-1 (2.0 equivalents) were added while maintaining the internal temperature below 20±5°C. After the addition was complete, container C was warmed to 30±5°C. Then, the contents of container A were transferred to container C over 4 hours while maintaining the internal temperature at 30±5°C. Container A was rinsed with toluene-2 ​​(0.5 L / kg) and transferred to container C. The contents of container C were stirred at 30°C for a further 3 hours. The contents of container C were cooled to 20±5°C. A solution of (s)-1-boc-3-methylpiperazine (1.2 equivalents) and N,N-diisopropylethylamine-2 (1.2 equivalents) in isopropyl acetate-1 (1.0 L / kg) was prepared in container D. While maintaining a batch temperature of 20±5°C, the solution from container D was added to container C (Note: exothermic reaction was observed). After the transfer was complete, container D was rinsed with additional dichloromethane (1.0 L / kg) and transferred to container C. The contents of container C were stirred at 20°C for a further 60 minutes. Then, a sodium bicarbonate solution [water-1 (15.0 L / kg + sodium bicarbonate (4.5 equivalents)]] was added to container C over 1 hour while maintaining an internal temperature of 20±5°C during the addition. The contents of container C were stirred for at least 12 hours, at which point the pipezoline (compound 6) product was isolated by filtration in a stirred filter dryer. The cake was washed with water-2 and -3 (5.0 L / kg x 2 times, each washing solution stirred for 15 minutes) and isopropyl acetate-2 and 3 (5.0 L / kg x 2 times, each washing solution stirred for 15 minutes). The cake was dried under nitrogen for 12 hours.

[0152] Acetone reslurrying (optional): Pipazoline (compound 6) and acetone (10.0 L / kg) were added to container E. The suspension was heated to 50°C for 2 hours. Water-4 (10.0 L / kg) was added to container E over 1 hour. After the addition of water was complete, the mixture was cooled to 20°C over 1 hour. The contents of container E were filtered to isolate the product, and the cake was washed with a 1:1 acetone / water mixture (5.0 L / kg). The cake was dried under nitrogen for 12 hours.

[0153] Process 6 [ka] General Notes: All equivalents and volumes are reported based on the amount of pipezoline added.

[0154] [Table 11]

[0155] To reactor A, add a solution of pipezoline (compound 6, 1.0 equivalent) in degassed water (6.5 L / kg), degassed 2-MeTHF (9.0 L / kg), and potassium acetate (2.0 equivalent). The resulting mixture is heated to 75 ± 5°C, and then a slurry of Pd(dpePhos)Cl2 (0.003 equivalent) in 2-MeTHF (0.5 L / kg) is added. Within 2 hours of catalyst addition, a freshly prepared solution of boroxine (compound 6A, 0.5 equivalent) in degassed moist 2-MeTHF (4.0 L / kg, KF > 4.0%) is added over a period of more than 1 hour but less than 2 hours, and after the addition is complete, rinse with further moist 2-MeTHF (0.5 L / kg). After the reaction is complete (less than 0.15 area% of pipazoline remains, typically less than 1 hour after the addition of boroxine), 0.2 wt% (0.002 kg / kg) of biaryl seed is added as a slurry to 0.02 L / kg of moist 2-MeTHF, and the resulting seed bed is aged for more than 60 minutes. Heptane (5.0 L / kg) is added over 2 hours at 75 ± 5°C. The batch is then cooled to 20 ± 5°C over 2 hours and aged for another 2 hours. The slurry is then filtered, and the cake is washed with 1 × 5.0 L / kg of water, 1 × 5.0 L / kg of 1:1 iPrOH:water, followed by 1 × 5.0 L / kg of 1:1 iPrOH:heptane (resuscitation washing: the cake is resuspended with a stirrer and allowed to stand before filtration). The cake (biaryl, compound 7) is then dried under vacuum using nitrogen sweeping. Note: If the reaction stops, further addition of catalyst and boroxine is required.

[0156] Step 7: Charcoal filtration for Pd removal [ka] General Notes: All equivalents and volumes are reported based on the amount of crude biaryl added.

[0157] [Table 12]

[0158] Place crude biaryl (1 equivalent) in a clean container A and add DCM (10 L / kg). While observing the dissolution, stir the contents at 22±5°C for >60 minutes. Remove 3 L of crude biaryl from container A. 2 The solution is passed through a bag filter and a carbon filter at a flux of less than / min / m, and the filtrate is collected in a clean container B. DCM rinse solution (1 L / kg) is added to container A, passed through a carbon filter, and collected in container B.

[0159] A solution sample is taken from the filtrate in container B to determine the IPC Pd content. The sample is concentrated into a solid and analyzed by ICP-MS. IPC: Pd ≤ 25 ppm relative to biaryl. a. If the Pd content of the first or second IPC sample exceeds 25 ppm relative to the biaryl, use 3 L of solution. 2 / min / m 2 The following steps involve passing the sample through a carbon filter a second time and rinsing with 1 L / kg of DCM. This is the IPC sample filtrate. b. If the Pd content exceeds 25 ppm after the third IPC, install a new carbon disc and adjust. Pass the biaryl filtrate through a refreshed carbon filter and wash with 1 L / kg DCM. IPC sample.

[0160] Distill and replenish to the appropriate concentration. Before proceeding to the Boc deprotection reaction in step 7, concentrate the filtrate to 4 L / kg or less, recover it, distill it, and replenish until the DCM reaches 5.25 ± 0.25 L / kg.

[0161] Process 7 [ka] General Notes: All equivalents and volumes are reported based on the amount of crude biaryl added.

[0162] [Table 13]

[0163] In reactor A, tert-butyl(3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridine-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidine-4-yl}-3-methylpiperazine-1-carboxylate (biaryl) (1.0 equivalent) and dichloromethane (5.0 L / kg) were added, and TFA (15.0 equivalents, 1.9 L / kg) was slowly added while maintaining the internal temperature at 20±5°C. The reaction mixture was stirred at 20±5°C for 4 hours.

[0164] Potassium carbonate (18.0 equivalents), water (20.0 L / kg), and NMP (1.0) were added to reactor B to produce a homogeneous solution. The reaction mixture from A was transferred to the potassium carbonate solution in B over 30 minutes while stirring at the maximum allowable speed of the apparatus (at a rate of approximately 0.24 L / kg / min). The mixture was stirred for a further 12 hours at 20±5°C.

[0165] The obtained slurry was filtered and rinsed with water (2 × 10 L / kg). The wet cake was dried for 24 hours to obtain 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperazin-1-yl]-(1M)-1-[4-methyl-2-(propan-2-yl)pyridine-3-yl]pyrido[2,3-d]pyrimidine-2(1H)-one (compound 8, DesBoc).

[0166] Process 8 [ka] General Notes: All equivalents and volumes are reported based on the amount of Des-Boc added.

[0167] [Table 14]

[0168] Add Des-Boc (compound 8, 1.0 equivalent) and NMP (4.2 L / kg) to container A under nitrogen, and slowly add TFA (1.0 equivalent) while maintaining Tr < 25°C. Age the mixture at 25°C (approximately 0.5 hours) until complete dissolution is observed. Then, filter the solution through a 0.45 micron filter into container B and wash with NMP (0.8 L / kg). Combine the filtrate and washings and cool to 0°C. Add acryloyl chloride (1.3 equivalents) to the resulting solution while maintaining the temperature below 10°C. Then, age the reaction mixture at 5 ± 5°C (approximately 1.5 hours) until the IPC is complete.

[0169] Preparation of aqueous disodium phosphate quench: Add disodium phosphate (3.0 equivalents) and water (15.0 L / kg) to container C. Age the mixture at 25°C until complete dissolution is observed. Warm the solution to 45±5°C. Prepare a seed slurry of AMG 510 (0.005 equivalents) in water (0.4 L / kg) and add it to container C while maintaining the temperature at 45±5°C.

[0170] The reaction mixture in container B is transferred to container C (quench solution) while maintaining the temperature at 45±5°C (approximately 1 hour). Container B is washed with a portion of NMP (0.5 L / kg). The resulting slurry is aged at 45±5°C for 2 hours, cooled to 20°C over 3 hours, aged at 20°C for a minimum of 12 hours, filtered, and washed with water (2 × 10.0 L / kg). The product is dried using nitrogen and vacuum to obtain crude AMG 510 (compound 9A).

[0171] Process 9 [ka] General Notes: All equivalents and volumes are reported based on the amount of crude AMG 510 used.

[0172] [Table 15]

[0173] To reactor A, 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridine-3-yl]-4-[(2S)-2-methyl-4-(propa-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidine-2(1H)-one (crude AMG 510) (1.0 equivalent), ethanol (7.5 L / kg), and water (1.9 L / kg) were added. The mixture was heated to 75°C and filtered into a clean reactor. The solution was cooled to 45°C, ground standard AMG 510 seeds (0.015 ± 0.005 kg / kg) were seeded, and the resulting slurry was aged for 30 minutes. While maintaining an internal temperature of >40°C, water (15.0 L / kg) was added over 5 hours, and the mixture was aged for a further 2 hours.

[0174] The mixture was cooled to 20°C over 3 hours, aged for 8 hours, and then the solid was recovered by filtration and washed with a mixture of ethanol (2.5 L / kg) and water (5.0 L / kg). The solid was dried using vacuum and nitrogen to obtain 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridine-3-yl]-4-[(2S)-2-methyl-4-(propa-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidine-2(1H)-one (AMG 510, compound 9).

[0175] A novel and advanced synthetic route for aniline (compound 2A) is disclosed below.

[0176] Synthesis procedure for 2-isopropyl-4-methylpyridine-3-amine (Scheme 1) [ka] Step 1: Reactor 1 was inactivated with nitrogen, and 1,2-dimethoxyethane (DME), aqueous K2CO3 solution (3 equivalents), Pd(PPh3)Cl2 (0.01 equivalent), 2-chloro-4-methylpyridine-3-amine (1.0 equivalent), and 4,4,5,5-tetramethyl-2-(propa-1-en-2-yl)-1,3,2-dioxaborolane (1.5 equivalents) were added. The reaction mixture was stirred and heated to 80°C until the reaction was complete by HPLC. The reaction mixture was then cooled to 25°C and filtered. The resulting solution was concentrated to remove the DME. Ether was added to the reactor, and the pH was adjusted to 2±1. The aqueous layer was removed and washed with dichloromethane. The pH of the aqueous layer was adjusted to 9-10 by adding aqueous NaOH solution. The aqueous layer was extracted with dichloromethane. The DCM layer was dried over Na2SO4, filtered, and concentrated at 40±5°C to obtain the crude product. Product. This material was diluted with EtOH and used directly in step 2.

[0177] Process 2 The EtOH solution of Pd / C (0.1 equivalents) and 4-methyl-2-(propa-1-en-2-yl)pyridine-3-amine from step 1 was placed in a nitrogen-inactivated pressure reactor. The reactor was evacuated and refilled with nitrogen (repeated 3 times), then evacuated and refilled with hydrogen (repeated 3 times). The reactor was stirred and the batch temperature was maintained at 45°C and the pressure at 7 bar for 16 hours. The reaction mixture was cooled to 25°C, evacuated, and refilled with nitrogen (repeated 3 times). The reaction mixture was filtered and concentrated to obtain crude 2-isopropyl-4-methylpyridine-3-amine, which was purified by distillation.

[0178] Synthesis of 2-isopropyl-4-methylpyridine-3-amine (Scheme 2) [ka] Step 1: Deactivate reactor 1 and add tetrahydrofuran. Start stirring and cool the batch to 5 °C. Add potassium tert-butoxide (3 equivalents) to reactor 1 little by little. Add a mixture of ethyl isobutyrate (1.0 equivalent) and acetonitrile (1 equivalent) to reactor 1 and age at ambient temperature for 16 hours. Cool the resulting reaction mixture to 0 °C and adjust the pH to 1.5 with concentrated hydrochloric acid. Extract the quenched reaction stream with dichloromethane. Wash the combined organic extracts with brine, dry over Na2SO4, filter. Concentrate the resulting solution and dilute with acetone. Add L-proline to reactor 1 and stir the resulting reaction mixture at 60 °C for 4 hours. Then, cool the reaction mixture to 25 °C, filter, and concentrate. Purify the residue by distillation to obtain 4-methyl-3-oxo-2-(propan-2-ylidene)pentanenitrile.

[0179] Step 2: Inactivate reactor 1 with nitrogen and add ethanol, (4-methyl-3-oxo-2-(propan-2-ylidene)pentanenitrile (1.0 equivalent), and DMF-DMA (1.2 equivalents). Stir the resulting mixture and warm to 70 °C for 4 hours. Cool the mixture to 25 °C and add NH4OH and NH4OAc to the reactor. Warm the reaction mixture to 70 °C, age for 16 hours, then cool to 40 °C and concentrate. Extract the mixture with dichloromethane, wash the combined organic extracts with brine, dry over Na2SO4, filter, and concentrate. Purify the residue by distillation to obtain 2-isopropyl-4-methylnicotinonitrile.

[0180] Step 3: Put 85% H2SO4 into reactor 1, and then add 2-isopropyl-4-methylnicotinonitrile (1.0 equivalent) little by little. Stir the reaction mixture, warm it at 90 °C for 16 hours, then cool it to 40 °C, and stop the reaction in a pre-cooled reactor 2 containing water at 0 - 5 °C. Adjust the pH to 8 with saturated Na2CO3 to obtain a precipitate of 2-isopropyl-4-methylnicotinamide. Isolate the crude product by filtration and add it back to reactor 3. Add EtOAc to reactor 3 and slurry the product at 80 °C for 1 hour. Then isolate the product by filtration and dry it to obtain 2-isopropyl-4-methylnicotinamide.

[0181] Step 4: Add an aqueous NaOH solution to reactor 1. Start stirring and cool the batch to 0 °C. Slowly add 2-isopropyl-4-methylnicotinamide (1.0 equivalent) and an aqueous sodium hypochlorite solution to the reactor little by little, and maintain the temperature below 10 °C during the addition. Then stir the reaction mixture at 5 °C for 1 hour and heat it to 70 °C for 16 hours. After completion, cool the reaction mixture to 25 °C and extract it with 50 / 50 EtOAc / THF. Wash the combined organic extracts with brine, dry them with Na2SO4, filter, and concentrate. Distill the obtained crude product to obtain pure 2-isopropyl-4-methylpyridin-3-amine.

[0182] A novel pyridine synthesis for the preparation of aniline (Scheme 3) is employed by employing a one-pot synthesis of a major nicotinonitrile intermediate from raw material chemicals. Michael addition of 4-methyl-3-oxopentanenitrile to crotonaldehyde proceeds under solvent-free conditions in the presence of a bulky secondary amine catalyst, yielding a cyclic tetrahydropyranlactol intermediate with high 1,4:1,2 selectivity (Chem.Commun. 2006, 4928). The tetrahydropyran intermediate is subjected to Ciufolini pyridine synthesis by treatment with hydroxylamine under thermal conditions, yielding the desired nicotinonitrile intermediate with complete regiocontrol (J.Am.Chem.Soc. 1996, 118, 12082). The present invention provides a direct stereodefining pathway to the core of aniline, aimed at reducing supply chain risks, lowering costs, and improving process immaturity for this major AMG 510 raw material. [ka]

[0183] Synthetic pathway of aniline via 2-isopropyl-4-methylnicotinonitrile: [ka] Description of the preparation process for 2-isopropyl-4-methylnicotinonitrile: Crotonaldehyde (1.10 equivalents) and (S)-α,α-bis[3,5-bis(trifluoromethyl)phenyl]-2-pyrrolidinemethanoltrimethylsilyl ether (0.05 equivalents) were added to container A. The contents of container A were stirred at 20°C (Tj) for up to 5 minutes. 4-methyl-3-oxopentannitrile (1.00 equivalent) was added to container A at a rate such that Tr < 25°C. The contents of container A were stirred at 20°C (Tj) for up to 12 hours. Then, acetonitrile (10 L / kg) was added to container A. The contents of container A were distilled until 5 L / kg of distillate was collected.

[0184] Hydroxylamine hydrochloride (2.00 equivalents) was added to container A, the contents of container A were heated to 80°C (Tj), stirred at 80°C (Tj) for 12 hours or less, and then cooled to 20°C (Tj).

[0185] Dichloromethane (5 L / kg) was added to container A, and then 5% sodium bicarbonate solution (5 L / kg) was added to container A at a rate of Tr < 25°C. The contents of container A were stirred at 20°C (Tj) for 30 minutes or less. Stirring was stopped, and the organic phase was discharged into container B.

[0186] Dichloromethane (5 L / kg) was added to container A, and the contents of container A were stirred at 20°C (Tj) for 30 minutes or less. Stirring was stopped, and the organic phase was discharged into container B.

[0187] The contents of container B were distilled until 10 L / kg of distillate was collected. Purification with silica gel eluted with 1:1 siRNA:heptane yielded 2-isopropyl-4-methylnicotinonitrile as a colorless oil.

[0188] The foregoing is merely illustrative of the present disclosure and is not intended to limit the invention to the disclosed uses. Modifications and alterations commonplace to those skilled in the art are intended to fall within the scope and nature of the invention as defined in the appended claims. All references, patents, applications and publications mentioned herein are incorporated herein by reference in their entirety as they are described herein.

Claims

1. Formula 2A 【Chemistry 1】 A method for producing a compound of the structure 【Chemistry 2】 A method comprising reacting a mixture containing a compound with palladium in a solvent in the presence of hydrogen.

2. 1,2-Dimethoxyethane (DME), K ２ CO ３ Aqueous solution (3 eq.), Pd(PPh ３ ) Cl ２ By reacting a mixture containing 2-chloro-4-methylpyridine-3-amine and 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane, the structure is obtained: 【Transformation 3】 The method according to claim 1, comprising producing a compound having [a certain characteristic].

3. Formula 2A: 【Chemistry 4】 A method for producing the compound, comprising an aqueous solution of NaOH, structure: 【Transformation 5】 A method comprising reacting a compound 36 having a compound with sodium hypochlorite with a mixture containing sodium hypochlorite.

4. structure: 【Transformation 6】 Compound 35 having H ２ SO ４ The method according to claim 3, comprising producing compound 36 by reacting with .

5. structure: 【Transformation 7】 Compound 34 having is reacted with DMF-DMA, NH ４ OH and NH ４ OOCCH ３ to obtain a structure: 【Transformation 8】 The method according to claim 4, comprising producing a compound 35 having the following properties.

6. structure: 【Chemistry 9】 Compound 33 having the following structure is reacted with potassium tert-butoxide, ethyl isobutyrate, acetonitrile and L-proline to obtain a compound with the following structure: 【Chemistry 10】 The method according to claim 5, comprising producing a compound 34 having the following properties.

7. a.) Crotonaldehyde and (S)-α,α-bis[3,5-bis(trifluoromethyl)phenyl]-2-pyrrolidinemethanoltrimethylsilyl ether; b. 4-methyl-3-oxopentannitrile; c.) Acetonitrile; and d.) Hydroxylamine hydrochloride Structure including a step of reacting a mixture: 【Chemistry 11】 A method for producing compound 35 having the properties of a compound.

8. Using the compound of formula 2A, formula 9: 【Chemistry 12】 The method according to claim 1, which produces a compound having

9. Using the compound of formula 2A, formula 9: 【Chemistry 13】 The method according to claim 3, which produces a compound having

10. Using the compound of formula 2A, formula 9: 【Chemistry 14】 The method according to claim 7, which produces a compound having

11. The method according to claim 8, further comprising mixing the compound of formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

12. The method according to claim 9, further comprising mixing the compound of formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

13. The method according to claim 10, further comprising mixing the compound of formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

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