Methods of making KRAS G12C inhibitors

Abstract

The present disclosure provides compounds of Formula I: # imgabs0 # or a pharmaceutically acceptable salt thereof, processes and intermediates useful for preparing compounds of Formula I, pharmaceutical compositions containing compounds of Formula I, and methods of using compounds of Formula I in the treatment of cancer.

Description

[0001] The present disclosure relates to compounds and pharmaceutically acceptable salts thereof that are useful for treating cancer.

[0002] WO 2021 / 118877 and US 2021 / 0179633 A1 (hereinafter referred to as "the '877 and '633 references") disclose certain KRas G12C inhibitors or salts thereof that can be used to treat cancer. Methods for preparing these compounds are also disclosed. Compounds made using the methods disclosed in the '877 and '633 references may contain impurities. These impurities may include Michael addition impurities that are difficult to remove.

[0003] It would therefore be useful to develop new methods and intermediates useful for preparing compounds of Formula I. Preferably, these new methods provide final compounds containing fewer impurities (such as Michael reaction-based impurities) and / or improved M:P atropisomer ratios. Summary of the invention

[0004] Disclosed herein are compounds of Formula I or pharmaceutically acceptable salts thereof obtainable by using newly developed intermediates and methods. Also disclosed herein are intermediates useful for preparing compounds of Formula I or pharmaceutically acceptable salts thereof. Also disclosed herein are methods for making compounds of Formula I or pharmaceutically acceptable salts thereof.

[0005] In one aspect, disclosed herein is a compound of Formula I or a pharmaceutically acceptable salt thereof

[0006]

[0007] It can be obtained by combining acryloyl chloride and 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazocin-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, the M atropisomer, in a continuous stirred tank reactor.

[0008]

[0009] wherein the Michael adduct impurity is less than 1% as measured by HPLC analysis.

[0010] In one aspect, the compound of formula I or a pharmaceutically acceptable salt thereof is prepared using the following intermediates or a pharmaceutically acceptable salt thereof,

[0011]

[0012] The intermediate can be obtained by reacting tert-butyl(13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazocine-2-carboxylate, tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate, potassium carbonate and a catalyst in a solvent, wherein the M:P atropisomer ratio is at least 4:1.

[0013] In another aspect, the compound of formula I or a pharmaceutically acceptable salt thereof is prepared using the following intermediates or a pharmaceutically acceptable salt thereof,

[0014]

[0015] The intermediate can be obtained by reacting (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylic acid tert-butyl ester, diacetate [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II), N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamic acid tert-butyl ester, a base and 1,1,1-tris(hydroxymethyl)ethane, wherein the M:P atropisomer ratio of the intermediate is at least 4:1.

[0016] Regarding intermediates that can be used to prepare compounds of Formula I or pharmaceutically acceptable salts thereof, disclosed herein are intermediate compounds of Formula Ii:

[0017]

[0018] or a pharmaceutically acceptable salt thereof,

[0019] in:

[0020] R is a protecting group.

[0021] In one aspect disclosed herein, the intermediate compound of Formula Ii is a compound of Formula Iia:

[0022]

[0023] or a pharmaceutically acceptable salt thereof.

[0024] In one aspect, disclosed herein is a compound of Formula Iia,

[0025]

[0026] or a pharmaceutically acceptable salt thereof,

[0027] It can be obtained by combining (3S)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester:phosphoric acid (1:1), base and 4-bromo-2,5-difluorobenzoic acid to give (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester.

[0028] Regarding intermediates that can be used to prepare compounds of Formula I or pharmaceutically acceptable salts thereof, disclosed herein are intermediate compounds of Formula IIi:

[0029]

[0030] or a pharmaceutically acceptable salt thereof,

[0031] in:

[0032] R is a protecting group.

[0033] In one aspect disclosed herein, the intermediate compound of formula IIi is a compound of formula IIia:

[0034]

[0035] or a pharmaceutically acceptable salt thereof.

[0036] In one aspect, disclosed herein is an intermediate compound of Formula IIia,

[0037]

[0038] or a pharmaceutically acceptable salt thereof,

[0039] It can be obtained by combining (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester with a cyclizing base to give (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester.

[0040] In one aspect, disclosed herein is an intermediate compound of formula IIIia,

[0041]

[0042] or a pharmaceutically acceptable salt thereof,

[0043] It can be obtained by combining (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepinatetraene-2-carboxylic acid tert-butyl ester with a chlorinating agent to give (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepinatetraene-2-carboxylic acid tert-butyl ester.

[0044] The present disclosure provides a compound selected from:

[0045]

[0046]

[0047] or a group consisting of pharmaceutically acceptable salts thereof.

[0048] The present disclosure also provides a compound selected from the group consisting of:

[0049]

[0050] or a group consisting of pharmaceutically acceptable salts thereof.

[0051] The present disclosure also provides a group of compounds selected from:

[0052]

[0053] The present disclosure provides a compound selected from the group consisting of:

[0054]

[0055] or a group consisting of pharmaceutically acceptable salts thereof.

[0056] The present disclosure provides a compound selected from the group consisting of:

[0057]

[0058] The present disclosure provides a compound selected from the group consisting of:

[0059]

[0060] or a group consisting of pharmaceutically acceptable salts thereof.

[0061] The present disclosure provides a compound wherein

[0062]

[0063] or a pharmaceutically acceptable salt thereof.

[0064] The present disclosure provides a compound wherein

[0065]

[0066] or a pharmaceutically acceptable salt thereof.

[0067] The present disclosure provides a compound selected from the group consisting of:

[0068]

[0069] or a group consisting of pharmaceutically acceptable salts thereof.

[0070] The present disclosure provides a compound wherein

[0071]

[0072] or a pharmaceutically acceptable salt thereof.

[0073] The present disclosure provides a compound wherein

[0074]

[0075] or a pharmaceutically acceptable salt thereof.

[0076] The present disclosure provides a compound selected from the group consisting of:

[0077]

[0078] or a group consisting of pharmaceutically acceptable salts thereof.

[0079] The present disclosure provides a compound wherein

[0080]

[0081] or a pharmaceutically acceptable salt thereof.

[0082] The present disclosure provides a compound wherein

[0083]

[0084] or a pharmaceutically acceptable salt thereof.

[0085] The present disclosure provides a compound selected from the group consisting of:

[0086]

[0087] or a group consisting of pharmaceutically acceptable salts thereof.

[0088] The present disclosure provides a compound wherein

[0089]

[0090] or a pharmaceutically acceptable salt thereof.

[0091] The present disclosure provides a compound wherein

[0092]

[0093] or a pharmaceutically acceptable salt thereof.

[0094] The present disclosure also provides a pharmaceutical composition comprising a compound according to any one of Examples 1-8 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

[0095] The present disclosure also provides a method for treating cancer, wherein one or more cells express KRas G12C mutant protein, comprising administering an effective amount of a compound according to any one of Examples 1-8 or a pharmaceutically acceptable salt thereof to a patient in need thereof. In various embodiments, the cancer is lung cancer such as advanced non-small cell lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, cervical cancer, endometrial cancer, ovarian cancer, bile duct cancer, or esophageal cancer. In a preferred embodiment, the cancer is advanced non-small cell lung cancer, pancreatic cancer, or colorectal cancer. In a more preferred embodiment, the cancer is non-small cell lung cancer.

[0096] In another form, the present disclosure comprises a method of treating cancer comprising administering an effective amount of a compound according to any one of Examples 1-8 or a pharmaceutically acceptable salt thereof to a patient in need thereof. In another embodiment, the cancer is non-small cell lung cancer. In another embodiment, the cancer is colorectal cancer. In yet another embodiment, the cancer is mutant pancreatic cancer. In another embodiment, the present disclosure comprises a method of treating cancers of other origins carrying a KRas G12C mutation.

[0097] The present disclosure also provides a method for treating a patient with cancer having a KRAS G12C mutation, comprising administering an effective amount of a compound according to any one of Examples 1-8 or a pharmaceutically acceptable salt thereof to a patient in need thereof.

[0098] The present disclosure also provides a method for regulating mutant KRas G12C enzyme in a patient in need thereof by administering a compound according to any one of Examples 1-8 or a pharmaceutically acceptable salt thereof. The method preferably comprises inhibiting human mutant KRas G12C enzyme.

[0099] The compounds of the present invention or their salts can be prepared by various procedures, some of which are described in the following "Preparation and Examples" and by the "Preparation and Examples" of the '877 and '633 references. The compounds of the present disclosure can be prepared according to the following "Preparation and Examples" by methods known and understood in the art and by the "Preparation and Examples" of the '877 and '633 references. Suitable reaction conditions for the steps of these "Preparation and Examples" are well known in the art, and appropriate substitution of solvents and co-reagents is within the technical scope of the art. Similarly, those skilled in the art will recognize that synthetic intermediates can be separated and / or purified by various known techniques as needed or expected, and various intermediates can usually be used directly in subsequent synthetic steps with little or no purification. As an example, the compounds of the "Preparation and Examples" can be separated, for example, by silica gel purification, directly separated by filtration or crystallization. In addition, the technician will recognize that in some cases, the order of introduction of the moieties is not important. As skilled chemists fully understand, the specific order of steps required to produce the compounds of the present disclosure depends on the relative instability of the specific compounds synthesized, the starting compounds, and the substituted moieties. Unless otherwise indicated, all substituents are as previously defined, and all reagents are well known and understood in the art.

[0100] The specific synthetic steps of each route can be combined in different ways, or combined with steps from different routes, to prepare the compounds or salts of the present disclosure. The products of each step in the following preparations can be recovered by conventional methods, including extraction, evaporation, precipitation, chromatography, filtration, trituration and crystallization.

[0101] The compounds of the present disclosure can be prepared in many ways known to those skilled in the art of organic synthesis. For example, the compounds of the present disclosure can be synthesized using the following methods and synthetic methods known in the field of synthetic organic chemistry or variations thereof as recognized by those skilled in the art. Preferred methods include, but are not limited to, those methods described below.

[0102] definition

[0103] Certain abbreviations are defined as follows: “APCI-MS” means atmospheric pressure chemical ionization mass spectrometry; “Boc” means tert-butyloxycarbonyl; “CDI” means 1,1′-carbonyldiimidazole; “CDMT” means 2-chloro-4,6-dimethoxy-1,3,5-triazine; “DCC” means 1,3-dicyclohexylcarbodiimide; “DIC” means 1,3-diisopropylcarbodiimide; “DIPEA” means N,N-diisopropylethylamine; “DMAc” means dimethylacetamide or DMA; “DMAP” means 4-dimethylaminopyridine; “DMF” means N,N-dimethylformamide; “DMSO” means dimethyl sulfoxide; “EDCI” means 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; “EtOH” means ethanol or ethyl alcohol; “HOAt” means 1-hydroxy-7-azobenzotriazole; “ "HBTU" refers to 3-[bis(dimethylamino)methylonium]-3H-benzotriazole-1-oxide hexafluorophosphate; "HOBt" refers to 1-hydroxybenzotriazole hydrate; "HPLC" refers to high performance liquid chromatography; "2-MeTHF" refers to 2-methyltetrahydrofuran; "MP(DSC)" refers to melting point measured by differential scanning calorimetry; "NCS" refers to N-chlorosuccinimide; "NMM" refers to N-methylmorpholine; "NMP" refers to N-methyl-2-pyrrolidone; "NMR" refers to nuclear magnetic resonance; "PG" refers to protecting group; "PyBOP" refers to (benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate); "PyBrOP" refers to bromo(tripyrrolidinyl)phosphonium hexafluorophosphate; "TEA" refers to triethylamine; "TFA" refers to trifluoroacetic acid; and "THF" refers to tetrahydrofuran.

[0104] Certain abbreviations are defined as follows: “ACN” means acetonitrile; “APCI-MS” means atmospheric pressure chemical ionization mass spectrometry; “Boc” means tert-butyloxycarbonyl; “CDI” means 1,1′-carbonyldiimidazole; “CDMT” means 2-chloro-4,6-dimethoxy-1,3,5-triazine; “DCC” means 1,3-dicyclohexylcarbodiimide; “DCM” means dichloromethane; “DIC” means 1,3-diisopropylcarbodiimide; “DIPEA” or “DIEA” means N,N-diisopropylethyl amine; "DMAc" or "DMA" refers to dimethylacetamide; "DMAP" refers to 4-dimethylaminopyridine; "DMF" refers to N,N-dimethylformamide; "DMSO" refers to dimethyl sulfoxide; "DMSO-d6" refers to deuterated dimethyl sulfoxide; "EDCI" refers to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; "EtOAc" refers to ethyl acetate; "EtOH" refers to ethanol or ethyl alcohol; "HATU" refers to O-(7-azabenzotriazol-1-yl)-N, N,N',N'-Tetramethyluronium hexafluorophosphate; "HOAt" refers to 1-hydroxy-7-azobenzotriazole; "HBTU" refers to 3-[bis(dimethylamino)methylium]-3H-benzotriazole-1-oxide hexafluorophosphate; "HOBt" refers to 1-hydroxybenzotriazole hydrate; "HPLC" refers to high performance liquid chromatography; "MeOH" refers to methanol; "2-MeTHF" refers to 2-methyltetrahydrofuran; "MP(DSC)" refers to melting point as measured by differential scanning calorimetry; "MT "BE" refers to methyl tert-butyl ether; "NCS" refers to N-chlorosuccinimide; "NMM" refers to N-methylmorpholine; "NMP" refers to N-methyl-2-pyrrolidone; "NMR" refers to nuclear magnetic resonance; "PG" refers to protecting group; "PyBOP" refers to (benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate); "PyBrOP" refers to bromo(tripyrrolidino)phosphonium hexafluorophosphate; "TEA" refers to triethylamine; "TFA" refers to trifluoroacetic acid; and "THF" refers to tetrahydrofuran.

[0105] A "base" is a molecule that acts as a proton acceptor, or a molecule that can neutralize an acid. Examples of bases include, but are not limited to, K2CO3.

[0106] "Cyclization base" is a base that facilitates cyclization. Examples of cyclization bases include sodium hydride, N,N-diisopropylethylamine (DIPEA or DIEA), triethylamine (TEA), cesium carbonate, diazabicycloundecene (DBU), sodium tert-butoxide, sodium tert-pentoxide, sodium tert-amylate, potassium tert-amylate or potassium tert-butoxide.

[0107] A "cyclization solvent" is a solvent that facilitates the cyclization reaction. Examples of cyclization solvents include DMF, NMP, DMAc, DMSO or THF.

[0108] A "chlorinating agent" is an agent that aids in chlorinating a molecule. Examples of chlorinating agents include trichloroisocyanuric acid or NCS.

[0109] The terms "continuous manufacturing" (hereinafter "CM"), "flow", and "flow chemistry" refer to the continuous supply of input materials into a manufacturing process (such as a continuous stirred tank reactor), the transformation of intermediate materials (in-process materials) within the manufacturing process, and the subsequent removal of output materials from the manufacturing process. These terms apply to the CM of new products (e.g., new drugs, generic drugs, biosimilars) and the conversion of batch manufacturing of existing products to CM. (See, e.g., FDA, "Q13 Continuous Manufacturing of Drug Substances and Drug Products, Guidance for Industry", March 2023).

[0110] The terms "batch", "batch manufacturing" and "batch chemistry" refer to a specific amount of material produced in a process or series of processes so that it is expected to be uniform within specified limits. Batch size can be defined by a fixed amount or by the amount produced in a fixed time interval. (See, e.g., FDA, "Q7 Good Manufacturing Practice Guidance for Active Pharmaceutical Ingredients, Guidance for Industry", September 2016).

[0111] A "catalyst" is a substance that increases the rate of a chemical reaction without undergoing any permanent chemical change itself.

[0112] The term "M:P atropisomer ratio" is the atropisomer ratio between the corresponding M and P atropisomers.

[0113] A "protecting group" is a reversibly formed derivative of an existing functional group in a molecule. Several types of protecting groups include alcohol protecting groups, amine protecting groups, carbonyl protecting groups, carboxylic acid protecting groups, phosphate protecting groups, and terminal alkyne protecting groups. Carboxylic acid protecting groups include protecting groups such as tert-butyl esters.

[0114] As used herein, the terms "treating" / "to treat" / "treatment" include slowing, alleviating or reversing the progression or severity of an existing symptom, disorder, condition, and may include specifically slowing the growth of cancerous lesions or the progression of abnormal cell growth and / or cell division.

[0115] As used herein, the term "PD-1" refers to programmed death receptor 1. As used herein, the term "PD-L1" refers to programmed death ligand 1.

[0116] As used herein, the term "CDK4 / CDK6 inhibitor" refers to any chemical that inhibits the function of CDK4 / CDK6.

[0117] As used herein, the term "EGFR inhibitor" refers to any chemical that inhibits the function of EGFR.

[0118] The term "ERK" refers to extracellular signal-regulated kinase. The term "ERK inhibitor" as used herein refers to any chemical that inhibits the function of ERK.

[0119] As used herein, the term "platinum agent" refers to any platinum-containing chemical that inhibits cancer.

[0120] As used herein, the term "antifolate" refers to any chemical that inhibits the function of folate. In one embodiment, the antifolate is pemetrexed.

[0121] As used herein, the term "Aurora A inhibitor" refers to any chemical that inhibits the function of Aurora A kinase.

[0122] As used herein, the term "SHP2 inhibitor" refers to any chemical that inhibits the function of SHP2.

[0123] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising: administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof simultaneously, separately or in sequential combination with a PD-1 or PD-L1 inhibitor at a dose of about 50 mg to about 200 mg, for the treatment of KRAS G12C mutant cancer. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof simultaneously, separately or in sequential combination with pembrolizumab in the treatment of KRAS G12C-mutant advanced NSCLC. In another embodiment, the PD-1 or PD-L1 inhibitor is pembrolizumab, wherein pembrolizumab is administered at 200 mg once every three weeks. In another embodiment, the PD-1 or PD-L1 inhibitor is nivolumab. In another embodiment, the PD-1 or PD-L1 inhibitor is cimiplimab. In another embodiment, the PD-1 or PD-L1 inhibitor is sindilimab. In another embodiment, the PD-1 or PD-L1 inhibitor is atezolizumab. In another embodiment, the PD-1 or PD-L1 inhibitor is avelumab. In another embodiment, the PD-1 or PD-L1 inhibitor is durvalumab. In another embodiment, the PD-1 or PD-L1 inhibitor is lodapilimab. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0124] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg, simultaneously, separately, or sequentially in combination with pembrolizumab in the treatment of KRASG12C-mutant advanced NSCLC, wherein pembrolizumab is administered intravenously at 200 mg once every three weeks. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0125] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof simultaneously, separately or in sequential combination with a CDK4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof at a dose of about 50 mg to about 200 mg. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof simultaneously, separately or in sequential combination with abemaciclib in the treatment of KRAS G12C-mutant advanced NSCLC at a dose of about 50 mg to about 200 mg. In another embodiment, the CDK4 / CDK6 inhibitor is abemaciclib, wherein abemaciclib is administered at 150 mg BID. In another embodiment, the CDK4 / CDK6 inhibitor is palbociclib. In another embodiment, the CDK4 / CDK6 inhibitor is ribociclib. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice daily. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0126] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of about 50 mg to about 200 mg simultaneously, separately or sequentially in combination with abemaciclib in the treatment of KRASG12C-mutant advanced NSCLC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0127] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof, simultaneously, separately or sequentially in combination with an EGFR inhibitor or a pharmaceutically acceptable salt thereof, at a dose of about 50 mg to about 200 mg. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof, simultaneously, separately or sequentially in combination with erlotinib in the treatment of KRAS G12C-mutant advanced NSCLC, at a dose of about 50 mg to about 200 mg. In another embodiment, the EGFR inhibitor is erlotinib, wherein erlotinib is administered once a day at 150 mg. In another embodiment, the EGFR inhibitor is afatinib. In another embodiment, the EGFR inhibitor is gefitinib. In another embodiment, the EGFR inhibitor is cetuximab. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice daily. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0128] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of about 50 mg to about 200 mg simultaneously, separately or sequentially in combination with erlotinib in the treatment of KRASG12C-mutant advanced NSCLC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0129] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of about 50 mg to about 200 mg simultaneously, separately, or sequentially in combination with cetuximab in the treatment of KRASG12C-mutant advanced CRC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0130] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof, simultaneously, separately or sequentially in combination with an ERK inhibitor or a pharmaceutically acceptable salt thereof, at a dose of about 50 mg to about 200 mg. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof, simultaneously, separately or sequentially in combination with LY3214996 in the treatment of KRAS G12C-mutant advanced NSCLC or CRC, at a dose of about 50 mg to about 200 mg. In another embodiment, the ERK inhibitor is LY3214996, wherein LY3214996 is administered at 400 mg twice a day. In another embodiment, the ERK inhibitor is LTT462. In another embodiment, the ERK inhibitor is KO-947. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice daily. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0131] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg simultaneously, separately, or sequentially in combination with LY3214996 in the treatment of KRASG12C-mutant advanced NSCLC or CRC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0132] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg simultaneously, separately, or sequentially in combination with a platinum agent. In another embodiment, the platinum agent is cisplatin. In another embodiment, the platinum agent is carboplatin. In another embodiment, the platinum agent is oxaliplatin. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0133] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose between about 50 mg and about 200 mg, which is used in combination with an antifolate simultaneously, separately or sequentially to treat KRAS G12C mutant cancer. In another embodiment, the antifolate is pemetrexed. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0134] In another aspect, disclosed herein is a method of treating a KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg and an Aurora A inhibitor or a pharmaceutically acceptable salt thereof. In another aspect, disclosed herein is a method of treating a KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg simultaneously, separately or sequentially in combination with (2R, 4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid: 2-methylpropan-2-amine (1:1) salt in the treatment of KRAS G12C-mutant advanced NSCLC. In another embodiment, the Aurora A inhibitor is an aminopyridine compound or a pharmaceutically acceptable salt thereof. In another embodiment, the Aurora A inhibitor is an Aurora A selective inhibitor or a pharmaceutically acceptable salt thereof. In another embodiment, the Aurora A inhibitor is alisertib as described in WO 2008 / 063525. In another embodiment, the Aurora A inhibitor is a pan Aurora inhibitor or a pharmaceutically acceptable salt thereof. In another embodiment, the Aurora A inhibitor is tozasertib as described in WO 2004 / 000833. In another embodiment, the Aurora A inhibitor is danasetib as described in WO 2005 / 005427. In another embodiment, the Aurora A inhibitor is (2R,4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid:

[0135]

[0136] or a pharmaceutically acceptable salt thereof. In one embodiment, the pharmaceutically acceptable salt is an amine salt. An example of an amine salt is an NH3 amine salt. Another example of an amine salt is a 2-methylpropane-2-amine salt.

[0137] In another aspect, disclosed herein is a method of treating a KRAS G12C mutant cancer comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof and (2R,4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid:amine (1:1) salt having the following structure:

[0138]

[0139] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof and (2R,4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1h-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid:2-methylpropan-2-amine (1:1) salt at a dose of between about 50 mg and about 200 mg, having the following structure:

[0140]

[0141] In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0142] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising: administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg simultaneously, separately or sequentially in combination with (2R, 4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1h-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid: 2-methylpropan-2-amine (1:1) salt in the treatment of KRAS G12C-mutant advanced NSCLC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0143] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg and a SHP2 inhibitor or a pharmaceutically acceptable salt thereof. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg simultaneously, separately or sequentially in combination with TNO155 in the treatment of KRAS G12C-mutant advanced NSCLC or CRC. In one embodiment, the SHP2 inhibitor is a type I SHP2 inhibitor or a type II SHP2 inhibitor. In another embodiment, the type I SHP2 inhibitor is PHPS1 or GS-493 or a pharmaceutically acceptable salt thereof. In another embodiment, the type I SHP2 inhibitor is NSC-87877 or NSC-117199 or a pharmaceutically acceptable salt thereof. In another embodiment, the type I SHP2 inhibitor is cefsulodin or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is JAB-3068 or JAB-3312 or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is RMC-4550 or RMC-4630 or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is SHP099, SHP244, SHP389, SHP394 or TNO155 or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is RG-6433 or RLY-1971 or a pharmaceutically acceptable salt thereof. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of formula I or a pharmaceutically acceptable salt thereof at a dose between about 50 mg and about 200 mg, and RMC-4630. In another aspect, disclosed herein is a method for treating a KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose between about 50 mg and about 200 mg, and JAB-3068. In another embodiment, the SHP2 inhibitor is BBP-398, IACS-15509 or IACS-13909 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is X37 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is ERAS-601 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is SH3809 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is HBI-2376 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is ETS-001 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is PCC0208023 or a pharmaceutically acceptable salt thereof.In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0144] In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising: administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg, simultaneously, separately, or sequentially in combination with TNO155 in the treatment of KRAS G12C-mutant advanced NSCLC or CRC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0145] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof, administered to a patient in need thereof at a dose of between about 50 mg and about 200 mg of the compound of Formula I or a pharmaceutically acceptable salt thereof, simultaneously, separately or sequentially in combination with a second therapeutic agent.

[0146] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating KRAS G12C mutant cancer, wherein the compound is administered simultaneously, separately or sequentially in combination with a second therapeutic agent at a dose between about 50 mg and about 200 mg, wherein the second therapeutic agent is selected from the group consisting of: a PD-1 inhibitor or a pharmaceutically acceptable salt thereof, a PD-L1 inhibitor or a pharmaceutically acceptable salt thereof, a CDK4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent or a pharmaceutically acceptable salt thereof, an antifolate or a pharmaceutically acceptable salt thereof, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, and a SHP2 inhibitor or a pharmaceutically acceptable salt thereof.

[0147] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof simultaneously, separately or in sequential combination with a PD-1 or PD-L1 inhibitor at a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof, for the treatment of KRAS G12C mutant cancer. In another embodiment, the PD-1 or PD-L1 inhibitor is pembrolizumab, wherein pembrolizumab is administered at 200 mg once every three weeks. In another embodiment, the PD-1 or PD-L1 inhibitor is nivolumab. In another embodiment, the PD-1 or PD-L1 inhibitor is cimiplimab. In another embodiment, the PD-1 or PD-L1 inhibitor is sindilimab. In another embodiment, the PD-1 or PD-L1 inhibitor is atezolizumab. In another embodiment, the PD-1 or PD-L1 inhibitor is avelumab. In another embodiment, the PD-1 or PD-L1 inhibitor is durvalumab. In another embodiment, the PD-1 or PD-L1 inhibitor is lodapilimab. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0148] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof in a dose of between about 50 mg and about 200 mg in combination with pembrolizumab simultaneously, separately, or sequentially in the treatment of KRAS G12C-mutant advanced NSCLC, wherein pembrolizumab is administered intravenously at 200 mg once every three weeks. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0149] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof simultaneously, separately or sequentially in combination with a CDK4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, at a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the CDK4 / CDK6 inhibitor is abemaciclib, wherein abemaciclib is administered at 150 mg BID. In another embodiment, the CDK4 / CDK6 inhibitor is palbociclib. In another embodiment, the CDK4 / CDK6 inhibitor is ribociclib. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0150] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof in a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof simultaneously, separately or sequentially in combination with abemaciclib in the treatment of KRAS G12C-mutant advanced NSCLC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0151] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof simultaneously, separately or in sequential combination with an EGFR inhibitor or a pharmaceutically acceptable salt thereof, in a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the EGFR inhibitor is erlotinib, wherein erlotinib is administered once a day at 150 mg. In another embodiment, the EGFR inhibitor is afatinib. In another embodiment, the EGFR inhibitor is gefitinib. In another embodiment, the EGFR inhibitor is cetuximab. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0152] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof in the treatment of KRAS G12C-mutant advanced NSCLC simultaneously, separately or sequentially in combination with erlotinib at a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0153] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof in a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof simultaneously, separately or sequentially in combination with cetuximab in the treatment of KRAS G12C-mutant advanced CRC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0154] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof simultaneously, separately or in sequential combination with an ERK inhibitor or a pharmaceutically acceptable salt thereof, at a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the ERK inhibitor is LY3214996, wherein LY3214996 is administered at 400 mg twice a day. In another embodiment, the ERK inhibitor is LTT462. In another embodiment, the ERK inhibitor is KO-947. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0155] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof in a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof simultaneously, separately or sequentially in combination with LY3214996 in the treatment of KRAS G12C-mutant advanced NSCLC or CRC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0156] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof simultaneously, separately or in sequential combination with a platinum agent at a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof. In another embodiment, the platinum agent is cisplatin. In another embodiment, the platinum agent is carboplatin. In another embodiment, the platinum agent is oxaliplatin. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0157] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which administers a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose of about 50 mg to about 200 mg to a patient in need thereof, which is used in combination with an antifolate simultaneously, separately or sequentially to treat KRAS G12C mutant cancer. In another embodiment, the antifolate is pemetrexed. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0158] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which administers a compound of Formula I or a pharmaceutically acceptable salt thereof and an Aurora A inhibitor or a pharmaceutically acceptable salt thereof at a dose of between about 50 mg and about 200 mg to a patient in need thereof. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof at a dose of between about 50 mg and about 200 mg in combination with (2R, 4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid:2-methylpropan-2-amine (1:1) salt simultaneously, separately or sequentially in combination in the treatment of KRAS G12C-mutant advanced NSCLC. In another embodiment, the Aurora A inhibitor is an aminopyridine compound or a pharmaceutically acceptable salt thereof. In another embodiment, the Aurora A inhibitor is an Aurora A selective inhibitor or a pharmaceutically acceptable salt thereof. In another embodiment, the Aurora A inhibitor is alisertib as described in WO 2008 / 063525. In another embodiment, the Aurora A inhibitor is a pan Aurora inhibitor or a pharmaceutically acceptable salt thereof. In another embodiment, the Aurora A inhibitor is tauzasertib as described in WO 2004 / 000833. In another embodiment, the Aurora A inhibitor is danasetib as described in WO 2005 / 005427. In another embodiment, the Aurora A inhibitor is (2R, 4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid or a pharmaceutically acceptable salt thereof. In another aspect, disclosed herein is a method of treating a KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof and (2R, 4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1H-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid:amine (1:1) salt at a dose between about 50 mg and about 200 mg. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0159] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of KRAS G12C-mutant advanced NSCLC, wherein the compound of Formula I or a pharmaceutically acceptable salt thereof is administered to a patient in need thereof in a dose of between about 50 mg and about 200 mg simultaneously, separately or sequentially in combination with (2R, 4R)-1-[(3-chloro-2-fluoro-phenyl)methyl]-4-[[3-fluoro-6-[(5-methyl-1h-pyrazol-3-yl)amino]-2-pyridinyl]methyl]-2-methyl-piperidine-4-carboxylic acid: 2-methylpropan-2-amine (1:1) salt. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0160] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, wherein a compound of Formula I or a pharmaceutically acceptable salt thereof and a SHP2 inhibitor or a pharmaceutically acceptable salt thereof are administered to a patient in need thereof at a dose of between about 50 mg and about 200 mg. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need thereof simultaneously, separately or in sequential combination with TNO155 in the treatment of KRAS G12C-mutant advanced NSCLC or CRC. In one embodiment, the SHP2 inhibitor is a type I SHP2 inhibitor or a type II SHP2 inhibitor. In another embodiment, the type I SHP2 inhibitor is PHPS1 or GS-493 or a pharmaceutically acceptable salt thereof. In another embodiment, the type I SHP2 inhibitor is NSC-87877 or NSC-117199 or a pharmaceutically acceptable salt thereof. In another embodiment, the type I SHP2 inhibitor is cefsulodin or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is JAB-3068 or JAB-3312 or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is RMC-4550 or RMC-4630 or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is SHP099, SHP244, SHP389, SHP394 or TNO155 or a pharmaceutically acceptable salt thereof. In another embodiment, the type II SHP2 inhibitor is RG-6433 or RLY-1971 or a pharmaceutically acceptable salt thereof. In another aspect, disclosed herein is a method for treating KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of formula I or a pharmaceutically acceptable salt thereof at a dose between about 50 mg and about 200 mg, and RMC-4630. In another aspect, disclosed herein is a method for treating a KRAS G12C mutant cancer, comprising administering to a patient in need thereof a compound of Formula I or a pharmaceutically acceptable salt thereof at a dose between about 50 mg and about 200 mg, and JAB-3068. In another embodiment, the SHP2 inhibitor is BBP-398, IACS-15509 or IACS-13909 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is X37 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is ERAS-601 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is SH3809 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is HBI-2376 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is ETS-001 or a pharmaceutically acceptable salt thereof. In another embodiment, the SHP2 inhibitor is PCC0208023 or a pharmaceutically acceptable salt thereof.In one embodiment, the dosage of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg, and about 200 mg.

[0161] In another aspect, disclosed herein is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament, which is administered to a patient in need thereof in a dose of about 50 mg to about 200 mg of a compound of Formula I or a pharmaceutically acceptable salt thereof simultaneously, separately or sequentially in combination with TNO155 in the treatment of KRAS G12C-mutant advanced NSCLC or CRC. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered once a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is administered twice a day. In one embodiment, the dose of the compound of Formula I or a pharmaceutically acceptable salt thereof is selected from the group consisting of about 50 mg, about 100 mg, about 150 mg and about 200 mg.

[0162] The compounds of the present disclosure can be synthesized in part by following the steps outlined in Scheme 1-1a below, which contains different orders of assembling intermediates or compounds. Starting materials can be purchased or made by known procedures reported in the literature or as shown below.

[0163] Schema 1

[0164]

[0165] In step 1 of Scheme 1, protected piperazine-2-ethanol (compound (1)) is coupled with 4-bromo-2,5-difluoro-benzoic acid (compound (2)) in amide bond formation using a coupling agent (such as CDMT) and an organic base (such as NMM) in a solvent system (such as acetonitrile and water) and an inorganic base (such as K2CO3). R is a protecting group developed for amino groups, such as carbamate and amide. Such protecting groups are well known and understood in the art, such as carbamate protecting groups, including allyloxycarbonyl, fluorenylmethoxycarbonyl or benzyloxycarbonyl. A common and preferred protecting group can be Boc. Those skilled in the art will recognize that there are many methods and reagents for amide formation resulting from the reaction of carboxylic acids and amines. For example, the reaction of an amine compound with an appropriate carboxylic acid in the presence of a coupling agent in the presence or absence of an organic base such as DIPEA or TEA can provide compound (3). Other coupling reagents include carbodiimides, such as DCC, DIC, EDCI or carbonyldiimidazoles, such as CDI. Amide coupling additives such as HOBt and HOAt can also be used to enhance the reaction. In addition, uronium or phosphonium salts of non-nucleophilic anions such as HBTU, HATU, PyBOP and PyBrOP can be used instead of more traditional coupling reagents. Additives such as DMAP can be used to enhance the reaction. Alternatively, the acyl chloride of compound (2) can be used in the presence of a base such as TEA or pyridine to obtain compound (3).

[0166] In step 2, the intramolecular cyclization of compound (3) is accomplished using a suitable base such as potassium tert-butoxide, sodium tert-amyloxide, sodium tert-butoxide, sodium tert-amyloxide, DIPEA, TEA, DBU, sodium hydride in a solvent such as DMF to give compound 4. Other possible solvents may be NMP, DMAc, DMSO and THF. This intramolecular cyclization of compound (3) to compound (4) can be performed by slowly adding a solution of compound (3) to an excess of base to minimize impurities derived from the intermolecular reaction.

[0167] In step 3, compound (4) can be chlorinated with a chlorinating agent such as trichloroisocyanuric acid or NCS in a solvent such as acetonitrile or DMF under acidic conditions using an acid such as TFA to give compound (5).

[0168] Figure 1a

[0169]

[0170] Scheme 1a illustrates the chiral synthesis of compound (5a). Compound (1a) can be prepared as described in Medicinal Chemistry route to 1, Development of an Alternative Route to the Bicyclic Piperazine, Retrosynthetic analysis of bicyclic piperazine core 2, and / or Coupling, cyclization, reduction, and Michael addition to afford Piperazine 24 in Org Proc Res Dev., 2011, 15(6). 1328-1335. Compounds (3a), (4a) and (5a) can be prepared as described in Scheme 1.

[0171] (S)-tert-Butyl 9-bromo-10-fluoro-12-oxo-1,2,4,4a,5,6-hexahydro-3H,12H-benzo[b]pyrazino[1,2-e][1,5]oxazocine-3-carboxylate was synthesized using a 7-step sequence starting from commercially available S-aspartic acid, which was the source of the stereocenter.

[0172] It is known that significant impurity exclusion can be achieved in the isolation of intermediates, including exclusion of the R-enantiomer. (S)-9-Bromo-10-fluoro-12-oxo-1,2,4,4a,5,6-hexahydro-3H,12H-benzo[b]pyrazino[1,2-e][1,5]oxazocine-3-carboxylic acid tert-butyl ester can be isolated by crystallization with significant impurity exclusion. Steps and Schemes are used to control process and reagent impurities.

[0173] The present disclosure provides a method for preparing an intermediate compound of formula IIa or a pharmaceutically acceptable salt thereof:

[0174]

[0175] It includes:

[0176] By using a cyclizing base, the intermediate compound of formula I:

[0177]

[0178] Or an intermediate compound of formula Ia:

[0179]

[0180] or a pharmaceutically acceptable salt thereof.

[0181] In one embodiment, the cyclizing base is selected from the group consisting of sodium hydride, N,N-diisopropylethylamine (DIPEA or DIEA), triethylamine (TEA), cesium carbonate, diazabicycloundecene (DBU), sodium tert-butoxide, sodium tert-amylate, sodium tert-amylate, potassium tert-amylate or potassium tert-butoxide.

[0182] In another embodiment, the preparation method further comprises a cyclization solvent. In one embodiment, the cyclization solvent is N,N-dimethylformamide (DMF).

[0183] The method of preparation, wherein the cyclization step is carried out at about 0°C. DETAILED DESCRIPTION

[0184] The KRAS G12C inhibitor of interest is 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, which has the following structure:

[0185]

[0186] This compound is disclosed as Example 35 in each of the '877 and '633 references. As disclosed therein, this compound exists as an atropisomer. Atropisomers can be separated. See Preparations 167 and 168 of the '877 and '633 references. The desired diastereomer is taught in Preparation 167 of the '877 and '633 references.

[0187] This desired diastereomer corresponds to the M atropisomer. The second diastereomer can be further elaborated to 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, the M atropisomer, which has the following structure:

[0188]

[0189] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer in each of the '877 and '633 references can be further elaborated to give Example 35, M atropisomer in each of the '877 and '633 references.

[0190] Example 35, M atropisomer is 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer (hereinafter referred to as “Formula I”) having the following structure:

[0191]

[0192] Compounds of Formula I are currently undergoing clinical trials (ClinicalTrials.gov Identifier: NCT04956640) to evaluate their effectiveness in treating patients with cancers that can be treated by inhibiting KRAS G12C. Compounds of Formula I can be used as a monotherapy, in combination with one or more other therapeutic agents, or as part of a neoadjuvant, adjuvant, advanced or metastatic therapy for the treatment of cancer. Such cancers include, but are not limited to, lung cancer, colorectal cancer, pancreatic cancer, bladder cancer, cervical cancer, endometrial cancer, ovarian cancer, bile duct cancer, or esophageal cancer.

[0193] Another example of a procedure of interest is Scheme 18 disclosed in each of the '877 and '633 references. As disclosed therein, Scheme 18 shows the coupling of compound 37 and compound 87 and the subsequent cyclization to give compound 88.

[0194] Another example of a procedure of interest is step D of Scheme 20 disclosed in each of the '877 and '633 references. As disclosed therein, step D shows the coupling between compound (97) and a partner to obtain a compound of formula I. The partner can be an acyl chloride, a carboxylic acid, or cyanogen bromide. In the case of using an acyl chloride, a suitable base such as TEA or DIEA is used in a solvent such as DCM. The acyl chloride can also be used with potassium carbonate as a base in a two-phase solvent system such as EtOAc, THF, and water. In the case where the partner is a carboxylic acid, the conditions include a suitable coupling agent (such as propylphosphonic anhydride) with a suitable base (such as DIEA) in a solvent (such as DMF). If the partner is cyanogen bromide, a suitable base (such as aqueous NaOH) is used in a solvent (such as DCM).

[0195] The compounds of the present disclosure or their salts can be prepared by various procedures, some of which are described in the following "Preparations and Examples". The specific synthetic steps of each route can be combined in different ways, or combined with steps from different routes, to prepare the compounds or salts of the present disclosure. The products of each step in the following preparation can be recovered by conventional methods, including extraction, evaporation, precipitation, chromatography, filtration, trituration and crystallization.

[0196] The compounds of the present disclosure can be prepared according to the following "Preparation and Examples" by methods known and understood in the art. Suitable reaction conditions for the steps of these "Preparation and Examples" are well known in the art, and appropriate substitutions of solvents and co-reagents are within the technical scope of the art. Similarly, those skilled in the art will recognize that synthetic intermediates can be separated and / or purified by various known techniques as needed or desired, and various intermediates can usually be used directly in subsequent synthetic steps with little or no purification.

[0197] For example, the compounds of the present disclosure can be synthesized using the following methods and synthetic methods known in the field of synthetic organic chemistry or variations thereof as recognized by those skilled in the art. As an example, the compounds of the "Preparation and Examples" can be separated, for example, by silica gel purification, by filtration or crystallization directly separated. In addition, the technician will recognize that in some cases, the order of introduction of the structural parts is not important. As skilled chemists fully understand, the specific order of steps required to produce the compounds of the present disclosure depends on the relative instability of the specific compounds synthesized, the starting compounds and the substituted parts. Unless otherwise indicated, all substituents are as previously defined, and all reagents are well known and understood in the art.

[0198] Also disclosed herein is a method for preparing an intermediate compound of Formula Iia or a pharmaceutically acceptable salt thereof, comprising combining (3S)-tert-butyl 3-(2-hydroxyethyl)piperazine-1-carboxylate: phosphoric acid (1:1), a base and 4-bromo-2,5-difluorobenzoic acid,

[0199]

[0200] To obtain (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester. In one embodiment, the combining step further comprises the step of adding CDMT, acetonitrile and NMM to 4-bromo-2,5-difluorobenzoic acid. In another embodiment, the combining step further comprises the step of adding K2CO3 and water to (3S)-3-(2-hydroxyethyl)piperazine-1-carboxylate: tert-butyl phosphate (1:1).

[0201] Also disclosed herein is a method for preparing an intermediate compound of formula IIia or a pharmaceutically acceptable salt thereof, which comprises cyclizing an intermediate compound of formula Ii or a pharmaceutically acceptable salt thereof, or an intermediate compound of formula Iia or a pharmaceutically acceptable salt thereof, by using a cyclizing base. In one embodiment, the cyclizing base is selected from the group consisting of sodium hydride, N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), cesium carbonate, diazabicycloundecene (DBU), sodium tert-butoxide, sodium tert-amyl alcoholate, sodium tert-amyl oxide, potassium tert-amyl alcoholate, and potassium tert-butoxide. In another embodiment, the preparation method further comprises a cyclization solvent. In yet another embodiment, the cyclization solvent is N,N-dimethylformamide (DMF). In yet another embodiment, the preparation method, wherein the cyclization step is carried out at about 0°C.

[0202] Also disclosed herein is a method for preparing a compound of Formula I, comprising combining acryloyl chloride and 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer in a continuous stirred tank reactor. In one embodiment, the combining step further comprises adding a feed A comprising a phosphate and water. In one embodiment, the phosphate comprises potassium dihydrogen phosphate and dipotassium hydrogen phosphate. In yet another embodiment, acryloyl chloride is included in feed B, wherein feed B comprises 2-methyltetrahydrofuran. In yet another embodiment, 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer is included in Feed C, wherein Feed C comprises 2-methyltetrahydrofuran and water. In yet another embodiment, the combining step further comprises pumping from the continuous stirred tank reactor to a second continuous stirred tank reactor. In yet another embodiment, the Michael adduct impurity is less than 1.5% as measured using the HPLC analysis described herein. In yet another embodiment, the Michael adduct impurity is less than 1% as measured using the HPLC analysis described herein. In yet another embodiment, the Michael adduct impurity is less than 0.75% as measured using the HPLC analysis described herein. In yet another embodiment, the Michael adduct impurity is less than 0.5% as measured using the HPLC analysis described herein. In yet another embodiment, the Michael adduct impurity is less than 0.4% as measured using the HPLC analysis described herein. In yet another embodiment, the Michael adduct impurity is less than 0.3% as measured using the HPLC analysis described herein. In yet another embodiment, the Michael adduct impurity is less than 0.25% as measured using the HPLC analysis described herein. In yet another embodiment, it further comprises the step of removing Michael adduct impurities.

[0203] For clarity, certain stereochemical centers are not specified and certain substituents have been eliminated in the following schemes, and it is not intended to limit the teaching of the schemes in any way. It is to be understood that all stereoisomers are included. Specific stereoisomers can be prepared by stereospecific synthesis using enantiomerically pure or enriched starting materials and / or reagents. Alternatively, enantiomers can be separated using methods known in the art, such as chiral chromatography or by converting enantiomers into diastereomeric salts, separating diastereomeric salts, converting diastereomeric salts into non-salt forms and separating enantiomers. Individual isomers, enantiomers and diastereomers may be separated or resolved by one of ordinary skill in the art at any convenient point in the synthesis of the disclosed compounds by methods such as selective crystallization techniques or chiral chromatography (see, e.g., J. Jacques et al., "Enantiomers, Racemates, and Resolutions", John Wiley and Sons, Inc., 1981 and EL Eliel and SH Wilen, "Stereochemistry of Organic Compounds", Wiley-Interscience, 1994).

[0204] In addition, the intermediates described in the following schemes contain many nitrogen or oxygen protecting groups. Variable protecting groups can be the same or different at each occurrence, depending on the specific reaction conditions and the specific conversion to be performed. Protection and deprotection conditions are well known to the technician and are described in the literature (see, for example, "Greene's Protective Groups in Organic Synthesis", 4th edition, Peter G.M. Wuts and Theodora W. Greene, John Wiley and Sons, Inc. 2007).

[0205] One skilled in the art will also recognize that the compounds of Formula I-IIa described herein, or pharmaceutically acceptable salts thereof, may be deuterated (wherein hydrogen may be replaced by deuterium), and that such molecules are within the scope of the compounds disclosed herein.

[0206] Preparation and Examples

[0207] Preparation 1

[0208] 4-Amino-3-chloro-2,5-difluoro-benzoic acid methyl ester

[0209]

[0210] The title compound was prepared in the same manner as in the '877 and '633 references; Preparation 31. ES / MS m / z: 222 [M+H] + .

[0211] Preparation 2

[0212] 3-Chloro-2,5-difluoro-4-iodo-benzoic acid methyl ester

[0213]

[0214] Cuprous iodide (22.7 g, 119 mmol), ACN (100 mL) and tert-butyl nitrite (17.7 mL, 149 mmol) were combined and stirred at room temperature for 45 minutes. A solution of 4-amino-3-chloro-2,5-difluoro-benzoic acid methyl ester (22.1 g, 100 mmol) in ACN (100 mL) was added. The reaction mixture was stirred at 40 ° C for 3 hours, cooled to room temperature, filtered through diatomaceous earth, and rinsed with DCM. The filtrate was concentrated in vacuo and purified by flash chromatography on silica gel (10% EtOAc in hexane) to obtain the title compound (17.8 g, 54%) as a white solid. 1 H NMR (CDCl3) δ7.60 (dd, J=5.6, 7.6Hz, 1H), 3.98 (s, 3H).

[0215] Preparation 3

[0216] 3-Chloro-2,5-difluoro-4-iodo-benzoic acid

[0217]

[0218] MeOH (50 mL) was added to a solution of 3-chloro-2,5-difluoro-4-iodo-benzoic acid methyl ester (16.8 g, 50.5 mmol) in THF (125 mL). The reaction mixture was cooled in an ice bath. 1N NaOH aqueous solution (150 mL, 150 mmol) was added in three portions. The reaction mixture was stirred at room temperature for 30 minutes and concentrated in vacuo to remove the organic solvent. The aqueous residue was cooled with crushed ice and the pH was adjusted to ~2 with 5N HCl aqueous solution (25 mL). The aqueous layer was extracted with EtOAc (3x). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to give the title compound (15.8 g, 98%). 1 H NMRδ (DMSO-d6) 13.91 (s, 1H), 7.68 (dd, J=5.8, 7.8Hz, 1H).

[0219] Preparation 4

[0220] (3R)-4-(3-Chloro-2,5-difluoro-4-iodo-benzoyl)-3-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester

[0221]

[0222] HATU (3.31 g, 8.53 mmol) was added to a solution of 3-chloro-2,5-difluoro-4-iodo-benzoic acid (2.99 g, 9.39 mmol) and DIEA (6.7 mL, 38 mmol) in THF (100 mL). The reaction mixture was stirred at room temperature for 1 hour. A solution of (3R)-3-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester (1.92 g, 8.52 mmol) in THF (5 mL) was added dropwise. The reaction mixture was stirred at room temperature for 14 hours and at 60 ° C for 3 hours, then diluted with EtOAc and water. The organic layer was washed with water, 1 N NaOH aqueous solution and brine. The water layer was extracted with EtOAc, which was washed with brine. The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (10-50% EtOAc in hexane) to obtain the title compound (3.82 g, 82%) as a white solid. ES / MS m / z( 35 Cl / 37 Cl) 461 / 463 [M-tert-butyl + H] + .

[0223] Preparation 5

[0224] (13aS)-9-Bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoxazoctate-2-carboxylic acid tert-butyl ester

[0225]

[0226] The title compound was prepared in the same manner as in the '877 and '633 references; Preparation of 117. ES / MS m / z ( 79 Br / 81 Br)407 / 409[M-tert-butyl+H] + .

[0227] Preparation 20

[0228] (3S)-4-(4-Bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester

[0229]

[0230] CDMT (802.8 g, 4.57 mol) was added to the reactor along with acetonitrile (20.0 L), and the mixture was cooled to 0°C. NMM (708.2 g, 7.00 mol) was added and the mixture was stirred at 0°C for 30 minutes. 4-Bromo-2,5-difluorobenzoic acid (794.8 g, 3.35 mol) was added.

[0231]

[0232] And the mixture was stirred at 0°C for 1 hour. In a separate container, K2CO3 (842g, 6.09mol), water (5.0L) and (3S)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester: phosphoric acid (1:1) (1000g, 3.05mol) were combined. The piperazine aqueous solution was added to the acetonitrile coupling solution over 5 minutes while maintaining an internal temperature of 0°C. The mixture was stirred at 0°C for 2.5 hours. The bottom layer was discharged from the reactor. The reaction mixture was stirred at 0°C for another 64 hours and then concentrated to 2L / kg. 2-MeTHF (10.0L) and n-heptane (3.0L) were added. 1M NaHCO3 aqueous solution (10.0L) was added and the mixture was stirred for 15 minutes. The layers were separated and the bottom layer was discharged. 1M NaHCO3 aqueous solution (10.0L) was added and the mixture was stirred for 15 minutes. The layers were separated and the bottom layer was discharged. Aqueous HCl (0.05M, 5.0L) was added, and the mixture was stirred for 15 minutes. The layers were separated, and the bottom layer was drained. Water (5.0L) was added, and the mixture was stirred for 15 minutes. The layers were separated, and the bottom layer was drained. The upper layer was concentrated to 2L / kg and 2-MeTHF (1.0L) was added. The mixture was concentrated to 2L / kg and 2-MeTHF (1.0L) was added. The mixture was concentrated to 2L / kg and DMF (2.7L) was added to give the title compound as a DMF solution (4.03kg, 31.2% assay, 2.80mol, 92% yield), which was used without further manipulation. The analytical sample was prepared by chromatography and exhibited the following spectral characteristics: APCI-MS measured values ​​471 and 473 (M+Na + ); 1H NMR (500 MHz, d6-DMSO+TFA, multiple rotational isomers exist) δ7.84 (m, 1H), 7.53 (br s, 1H), 4.71 (br s, 0.5H), 4.33–4.27 (m, 0.5H), 4.12–3.72 (m, 2H), 3.65 (br s, 0.5H), 3.47 (br, 1H), 3.38–3.11 (m, 2H), 3.11–2.56 (m, 2.5H), 1.68 (m, 2H), 1.39 (s 9H).

[0233] Preparation 21

[0234] (13aS)-9-Bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoxazoctate-2-carboxylic acid tert-butyl ester

[0235]

[0236] Sodium tert-amyl alcohol (462g, 4.20mol) was added to the reactor together with DMF (9.1L). The mixture was stirred to form a solution and cooled to 0°C. A DMF solution of (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester (4.03kg, 31.2% assay, 2.80mol) was added at 0°C over 8 hours. The total mass of the product solution was 12.95kg at this time. A portion of the solution (1.30kg) was placed in a separate reactor. Water (2.6kg) was added to a clean reactor. 1.30kg of the product solution was added to water over 3 hours at 25°C. The mixture was stirred at 25°C for 64 hours. The resulting solid was separated by filtration and washed with a mixture of DMF (0.3L) and water (0.3L). The crude solid was added to the reactor together with EtOH (628mL). The mixture was heated to 55 ° C and stirred for 30 minutes. Water (314 mL) was added over 2 hours and the mixture was kept at 55 ° C for another 30 minutes. The slurry was cooled to 20 ° C over 2 hours and then kept at 20 ° C for 2 hours. The resulting solid was separated by filtration and washed with a mixture of water (60 mL) and EtOH (60 mL). The product was dried under vacuum at 50 ° C for 16 hours to give the title compound (98.2 g, 73% yield). APCI-MS found 451 and 453 (M+Na +1 ); 1H NMR(400MHz,d6-DMSO-)δ7.32(d,J=6.0Hz,1H),7.28(d,J=9.2Hz,1H),4.24–4.18(m,2H),4.05–3.91( m,2H),3.87–3.76(m,2H),3.06(br,1H),2.95–2.80(brm,2H),2.06(m,1H),1.76(m,1H),1.04(s,9H).

[0237] Alternative Synthesis of Preparation 5

[0238] (13aS)-9-Bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoxazoctate-2-carboxylic acid tert-butyl ester

[0239]

[0240] Tert-butyl (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylate (2.00 g, 4.66 mmol) was added to the reactor along with acetonitrile (20 mL), and the mixture was cooled to 5°C. Trichloroisocyanuric acid (1.08 g, 4.66 mmol) was added and the mixture was stirred at 5°C for 5 minutes. Phosphoric acid solution (0.96 mL, 14.0 mmol; stock solution was 1.92 mL phosphoric acid in 4.0 mL acetonitrile) was added over 2 minutes, and the mixture was stirred at 5°C for 23 hours. Aqueous K2CO3 solution (40%, 20 mL) was added and the mixture was stirred at 5°C for 30 minutes, warmed to 20°C, and stirred at 20°C for 2 hours. Ethyl acetate (20 mL) and water (20 mL) were added and the mixture was stirred until the solid dissolved. Solids reappeared after standing for 1 hour. Aqueous NaHSO3 solution (5%, 10 mL) was added and the mixture was stirred overnight. Aqueous K2CO3 solution (40%, 4 mL) and di-tert-butyl dicarbonate (0.20 g, 0.93 mmol) were added and the mixture was stirred overnight. The mixture was concentrated to 2-3 volumes and isopropanol (4 volumes) was added. The mixture was concentrated to 2-3 volumes and isopropanol (4 volumes) was added. The mixture was concentrated to 2-3 volumes and isopropanol (2 volumes) was added. The mixture was stirred at 57°C. Water (4 volumes) was added over 1 hour, and the mixture was stirred at 57°C for 2 hours. The mixture was cooled and stirred at 20°C overnight, filtered, and washed with 1:2 isopropanol:water (3 volumes). The product was dried under vacuum at 50°C for 3 hours to give the title compound (1.85 g, 86% yield).

[0241] Preparation 6

[0242] (4aR)-7-Chloro-9-fluoro-8-iodo-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -3-tert-Butyl formate

[0243]

[0244] Tert-butyl (3R)-4-(3-chloro-2,5-difluoro-4-iodo-benzoyl)-3-(hydroxymethyl)piperazine-1-carboxylate (4.59 g, 8.88 mmol) in DMF (150 mL) was cooled to 0 °C. Sodium hydride (0.89 g, 22 mmol, 60% by mass in paraffin oil) was added. After 4 hours at 0 °C, the reaction mixture was quenched with saturated aqueous sodium bicarbonate and diluted with water and diethyl ether. The aqueous layer was extracted with diethyl ether (4x). The combined organic extracts were washed with water (3x) and brine, dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (15-40% EtOAc in hexanes) to give the title compound (3.82 g, 82%) as a white solid. ES / MS m / z( 35 Cl / 37 Cl) 441 / 443 [M-tert-butyl + H] + .

[0245] Preparation 7

[0246] N-(4-Bromo-3-cyano-7-fluoro-benzothiophen-2-yl)carbamic acid tert-butyl ester

[0247]

[0248] The title compound was prepared in the same manner as in the '877 and '633 references; Preparation 14. ES / MS m / z ( 79 Br / 81 Br)315 / 317[M-tert-butyl+H] + .

[0249] Preparation 8

[0250] tert-Butyl N-[3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiophen-2-yl]carbamate

[0251]

[0252] A mixture of tert-butyl N-(4-bromo-3-cyano-7-fluoro-benzothiophene-2-yl)carbamate (3.10 g, 8.35 mmol), bis(pinacol)diboron (21.0 g, 82.7 mmol) and potassium acetate (2.50 g, 25.5 mmol) in 1,4-dioxane (50 mL) was purged with nitrogen (direct bubbling) for 10 minutes. 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II) dichloromethane complex (1.0 g, 1.2 mmol) was added. The reaction flask was sealed and heated at 80-85 ° C for 3.5 hours. The reaction mixture was filtered through diatomaceous earth. The filtrate was diluted with water and extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (10%-50% (20% acetone / DCM) / hexanes) to give the title compound (4.45 g, 87%). ES / MS m / z 363 [M-tert-butyl+H] + .

[0253] Preparation 9

[0254] tert-Butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborin-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate

[0255]

[0256] The title compound was prepared in the same manner as the '877 and '633 references; Preparation 15. 1 H NMR(DMSO-d6)δ11.6(s,1H),7.61(m,1H),7.20(m,1H),3.78(s,4H),1.54(s,9H),1.03(s,6H).

[0257] Preparation 10 and 11

[0258] (13aS)-9-[2-(tert-Butyloxycarbonylamino)-3-cyano-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate-2-carboxylic acid tert-butyl ester, P and M atropisomers

[0259]

[0260] The title compound was prepared in the same manner as in the '877 and '633 references; Preparation 167. The mixture of atropisomers was separated by silica gel flash chromatography (0-30% acetone in hexanes). Preparation 10 (P atropisomer) was the first compound to elute from the column. Preparation 11 (M atropisomer) was the second compound to elute from the column. For both, ES / MS m / z ( 35 Cl / 37 Cl)657 / 659[M+H] + .

[0261] Preparation 12 and 13

[0262] (13aS)-9-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoxazoctate-2-carboxylic acid tert-butyl ester, P and M atropisomers

[0263]

[0264] (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazo ... A mixture of 1,4-dioxane (215 mL) was added to 1,4-dioxane (215 mL) which had been purged with nitrogen (bubbled directly) for 30 minutes. The reaction flask was sealed and heated at 105° C. for 14 hours. Additional tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate (3.54 g, 8.75 mmol) and dichloro[(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) (0.97 g, 1.12 mmol) were added. The reaction flask was sealed and heated at 105°C for 14 hours.

[0265] The reaction mixture was filtered through celite and rinsed with EtOAc. The filtrate was concentrated in vacuo and diluted with EtOAc, water and brine. The aqueous layer was extracted with EtOAc. The combined organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel flash chromatography (10-40% acetone in hexanes). Preparation 12 (P-retarsoisomer) was eluted first, followed by Preparation 13 (M-retarsoisomer). The impure fraction of the P-retarsoisomer was further purified by silica gel flash chromatography. This produced two title compounds (P-retarsoisomer, 0.35 g, 2%; M-retarsoisomer, 5.6 g, 38%). For both, ES / MS m / z 619 [M-tert-butyl+H] + .

[0266] Catalyst screening for the preparation of 12 and 13

[0267] (13aS)-9-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoxazoctate-2-carboxylate tert-butyl ester, P and M atropisomers

[0268]

[0269] In a glove box, a mixture of (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester (0.025 g, 0.054 mmol), N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborin-2-yl)-7-fluoro-benzothiophen-2-yl]carbamic acid tert-butyl ester (0.030 g, 0.074 mmol), potassium carbonate (0.022 g, 0.16 mmol) and catalyst (0.0081 mmol) was added to 1,4-dioxane (1 mL) in a vial. The vial was sealed and heated at 105 °C for 19 hours and then cooled to room temperature. Monitoring the reaction by LCMS showed the following reaction progress and atropisomer ratios:

[0270] Table 1:

[0271]

[0272] Alternative Synthesis of Preparation 13

[0273] (13aS)-9-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoxazoctate-2-carboxylic acid tert-butyl ester, M atropisomer

[0274]

[0275] In a glove box, palladium(II) acetate (0.119 g, 0.528 mmol), (S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (0.364 g, 0.580 mmol) and toluene (25 mL) were added to a small flask, which will make [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) diacetate. The mixture was capped and stirred for 1 hour. (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylic acid tert-butyl ester (0.500 g, 1.06 mmol) and aqueous potassium carbonate (2 mL, made from 0.241 g potassium carbonate in 2 mL water) were added. The mixture was heated at 60°C for 6 hours. The organic and aqueous layers were separated. The organic layer was dried over magnesium sulfate and filtered. The solid was rinsed with toluene (25 mL) which was added to the original filtrate.

[0276] To the second flask was added tert-butyl (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylate (9.51 g, 20.16 mmol) and cesium carbonate (31.1 g, 95.0 mmol). The flask was cycled through vacuum and nitrogen four times.

[0277] To the third flask was added tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate (10.9 g, 25.3 mmol), 1,1,1-tris(hydroxymethyl)ethane (1.69 g) and THF (50 mL). The mixture was stirred for 25 minutes.

[0278] To the second flask was added the ligand-catalyst filtrate and THF (50 mL). The mixture was heated at 60 °C under nitrogen. The contents of the third flask were added dropwise (6 mL / hour). After a total of 10 hours of heating, HPLC analysis showed complete conversion of the bromide starting material to a product with a 4.36:1 M:P atropisomer ratio.

[0279] After a total of 12 hours of heating, the mixture was allowed to cool to room temperature. Water (100 mL) was added, and the organic and aqueous layers were separated. 20% brine (50 mL) was added, and the organic and aqueous layers were separated. A portion (-20%) of the organic layer (26.2 g) was distilled to -10 mL, diluted with isopropyl acetate (20 mL), distilled to -10 mL, diluted with isopropyl acetate (20 mL), and distilled to -10 mL. The resulting suspension was filtered and dried under vacuum to give the title compound (1.85 g). HPLC analysis showed 98.23% M atropisomer and 0.76% P atropisomer.

[0280] Preparation 14 and 15

[0281] (4aR)-8-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -tert-Butyl 3-carboxylate, P and M atropisomers

[0282]

[0283] (4aR)-7-Chloro-9-fluoro-8-iodo-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine A solution of tert-butyl-3-formate (0.50 g, 1.01 mmol), tert-butyl N-[3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiophen-2-yl]carbamate (0.63 g, 1.51 mmol) and potassium phosphate (0.50 g, 2.36 mmol) in water (2 mL) and 1,4-dioxane (10 mL) was purged with nitrogen (bubbled directly) for 10 minutes. 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride (0.20 g, 0.30 mmol) was added. The reaction flask was sealed and heated at 70 °C for 3 hours.

[0284] (4aR)-7-Chloro-9-fluoro-8-iodo-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine A solution of tert-butyl-3-formate (0.20 g, 0.40 mmol), tert-butyl N-[3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiophen-2-yl]carbamate (0.24 g, 0.57 mmol) and potassium phosphate (0.20 g, 0.96 mmol) in water (2.5 mL) and 1,4-dioxane (8 mL) was purged with nitrogen (bubbled directly) for 10 minutes. 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride (0.080 g, 0.12 mmol) was added. The reaction flask was sealed and heated at 70°C for 5 hours.

[0285] The two reaction mixtures were combined, filtered through celite, and rinsed with EtOAc. The filtrate was diluted with MTBE and saturated aqueous sodium bicarbonate. The organic extract was washed with brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel flash chromatography (0-30% acetone in hexanes). Preparation 14 (P atropisomer) eluted first, followed by Preparation 15 (M atropisomer). The impure fraction of the P atropisomer was further purified by silica gel flash chromatography (0-100% EtOAc in hexanes). This produced the two title compounds (P atropisomer, 0.17 g, 17%; M atropisomer, 0.21 g, 21%). For both, ES / MS m / z ( 35 Cl / 37 Cl) 605 / 607 [M-tert-butyl + H] + .

[0286] Preparation 16

[0287] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile, P-atropisomer

[0288]

[0289] TFA (4 mL) was added to a solution of (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylic acid tert-butyl ester, P-atropisomer (0.98 g, 1.5 mmol) in DCM (5 mL). The reaction mixture was stirred at room temperature for 3 hours, concentrated in vacuo, and purified by silica gel flash chromatography (4-10% 7N ammoniated MeOH in DCM) to give the title compound (0.57 g, 84%). ES / MS m / z ( 35 Cl / 37 Cl)457 / 459[M+H] + .

[0290] Preparation 17

[0291] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile, M atropisomer

[0292]

[0293] The title compound was prepared in the same manner as in the '877 and '633 references; Preparation 187. ES / MS m / z ( 35 Cl / 37 Cl)457 / 459[M+H] + .

[0294] Preparation 18

[0295] 4-[(4aR)-7-Chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, P-atropisomer

[0296]

[0297] TFA (2 mL) was added to (4aR)-8-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -3-tert-Butyl formate, P-atrosomer (0.018 g, 0.027 mmol) in DCM (2 mL). The reaction mixture was stirred at room temperature for 30 minutes, concentrated in vacuo, and purified by flash chromatography on silica gel (0-10% MeOH in DCM, then 0-10% 7N ammoniated MeOH in DCM) to give the title compound (0.013 g, 100%). ES / MS m / z ( 35 Cl / 37 Cl)461 / 463[M+H] + .

[0298] Preparation 19

[0299] 4-[(4aR)-7-Chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer

[0300]

[0301] TFA (2.5 mL) was added to (4aR)-8-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -tert-Butyl 3-formate, M atropisomer (0.20 g, 0.31 mmol) in an ice-cooled solution of DCM (2.5 mL). The reaction mixture was stirred at room temperature for 2 hours, concentrated in vacuo, and purified by silica gel flash chromatography (0-10% 7N ammoniated MeOH in DCM) to give the title compound (0.13 g, 89%). ES / MS m / z ( 35 Cl / 37 Cl)461 / 463[M+H] + .

[0302] Preparation 22

[0303] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer

[0304]

[0305] (13aS)-9-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylic acid tert-butyl ester, M atropisomer (100.0 g, 148.1 mmol) was added to a reactor with EtOH (300 mL). A solution of HCl in EtOH (30%, 216 g, 1.78 mol) was added over 30 minutes with stirring. The mixture was heated to 40 °C for 20 hours and then cooled to 0 °C with stirring. A mixture of KOH (118.8 g, 1.80 mol) in water (1.0 L) was gradually added with stirring. EtOH was removed by distillation. The residue was extracted with 2-MeTHF (1.0 L) and extracted again with 2-MeTHF (500 mL). The organic extracts were combined, washed with saturated aqueous NaCl solution (3×0.5 L), and concentrated to ˜200 mL. Acetonitrile (1.0 L) was added dropwise. The mixture was stirred and cooled to 0° C. for 1 hour. The solid was collected by filtration, washed with acetonitrile (200 mL), and dried under vacuum to give the title compound (66.4 g, 85% yield) as a white solid. APCI-MS 475 (M+H + ).

[0306] Examples 1 and 2

[0307] (13aS)-9-(2-Amino-7-fluoro-1,3-benzothiazol-4-yl)-8,10-dichloro-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-6-one, P and M atropisomers

[0308]

[0309] The title compound was prepared in the same manner as in the '877 and '633 references; Example 1. The mixture of atropisomers was used IC, 4.6x150mm, 40% EtOH / CO2, 5mL / min, 225nm separation. Example 1 (P atropisomer) is the first compound to leave the column. Example 2 (M atropisomer) is the second compound to leave the column. For both, ES / MS m / z ( 35 Cl / 37 Cl)521 / 523[M+H] + .

[0310] Example 3

[0311] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile, P-atropisomer

[0312]

[0313] A solution of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile, P-atropisomer (0.115 g, 0.252 mmol) and DIEA (0.20 mL, 1.2 mmol) in DCM (2 mL) was cooled to -78 °C. Acryloyl chloride (0.02 mL, 0.2 mmol) was added. After 5 minutes, it was diluted with a small amount of 2-propanol and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (20-100% acetone in hexanes) to give the title compound (0.091 g, 71%). ES / MS m / z( 35 Cl / 37 Cl)511 / 513[M+H] + .

[0314] Example 4

[0315] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile, M atropisomer

[0316]

[0317] The title compound was prepared in the same manner as in the '877 and '633 references; Example 34. ES / MS m / z ( 35 Cl / 37 Cl)511 / 513[M+H] + .

[0318] Example 5

[0319] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, P-atropisomer

[0320]

[0321] TFA (1 mL) was added to a solution of (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylic acid tert-butyl ester, P-atropisomer (0.025 g, 0.037 mmol) in DCM (1 mL). The reaction mixture was stirred at room temperature for 30 minutes and then concentrated in vacuo to give the crude deprotected material.

[0322] The residue was dissolved in DCM (1 mL) and DIEA (0.013 mL, 0.074 mmol) was added. The solution was cooled to -78 °C. Acryloyl chloride (0.0030 mL) in DCM (1 mL) was added. After 30 minutes, it was concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (20-80% acetone in hexanes) to give the title compound (0.012 g, 61%). ES / MS m / z ( 35 Cl / 37 Cl) 529 / 531[M+H] + .

[0323] Example 6

[0324] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer

[0325]

[0326] HCl gas is bubbled into (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophene-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazine [2,1-d] [1,5] benzoxazopyrazine octate-2-carboxylic acid tert-butyl ester, M atropisomer (0.786g, 1.17mmol) in DCM (12mL) and 2-propanol (12mL) ice-cooled solution for 5 minutes. The reaction mixture is stirred at room temperature for 5 hours and then cooled in an ice bath. HCl gas is bubbled into the reaction mixture for 5 minutes. The reaction mixture is stirred at room temperature for 14 hours and then concentrated in vacuo. The residue is diluted twice with n-heptane and concentrated in vacuo. MTBE (50mL) is added. The mixture is stirred at room temperature for 10 minutes and then filtered to obtain the deprotected material as dihydrochloride.

[0327] The dihydrochloride salt was dissolved in water (12 mL). 2-Methyltetrahydrofuran (12 mL) was added. A solution of potassium carbonate (0.81 g, 5.82 mmol) in water (12 mL) was added. The mixture was stirred vigorously while being cooled in an ice bath. Acryloyl chloride (0.10 mL, 1.28 mmol) in 2-methyltetrahydrofuran (5 mL) was added dropwise. After 5 minutes, it was diluted with brine and 2-methyltetrahydrofuran. The aqueous layer was extracted with 2-methyltetrahydrofuran (2x). The combined organic extracts were washed with water, dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (20-30% B / A; A: 3:1 EtOAc: Hexane; B: 4:1 EtOAc: MeOH) to give the title compound (0.409 g, 66%). ES / MS m / z( 35 Cl / 37 Cl) 529 / 531[M+H] + .

[0328] Alternative Synthesis of Example 6: Flow Chemistry

[0329] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer

[0330]

[0331] To a continuous stirred tank reactor (105 mL, 15° C.) were added feed A [monopotassium phosphate (25.02 g), dipotassium phosphate (2.61 g) and water (to 600 ml volume); pH 5.8; flow rate 4.73 g / min], feed B [acryloyl chloride (13.15 g, 1.15 eq) and 2-methyltetrahydrofuran (6 vol); flow rate 2.48 g / min] and feed C [4-[(13 aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer (60 g, 0.126 mol), 2-methyltetrahydrofuran (6 volumes) and water (0.2 volumes); flow rate 2.97 g / min]. The mixture in the reactor was pumped to a second continuously stirred tank reactor (105 mL, 15° C.) at a flow rate of 10.5 g / min and then collected in a separate receiving bottle. HPLC analysis conditions ("HPLC analysis") comprised an Inertsil ODS-3V column (4.6 x 250 mm; 5 mm) eluting with 20% to 95% ACN / 0.1% H3PO4 in water using an Agilent 1260 chromatograph with UV detector. HPLC analysis showed 0.33% impurity, 4-[(13aS)-2-[3-[(13aS)-9-(2-amino-3-cyano-7-fluoro-benzothiophene-4-yl)-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazaoctatetraen-2-yl]-3-oxo-propyl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazaoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M,M atropisomer,

[0332]

[0333] "MA impurity" as shown in Table 2. This MA impurity is a Michael adduct in which unreacted [4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer reacts with the compound of Formula I and thereby forms a dimer. Trace amounts of the corresponding M,P atropisomer dimer may be present. The lower percentage of this impurity in the flow reaction is surprising and unexpected relative to the higher percentage of this impurity in the comparative batch reaction. The final mixture layers were separated. The aqueous phase was extracted with 2-methyltetrahydrofuran (5 volumes). The combined organic phases were washed twice with aqueous sodium bicarbonate (5 weight %, 10 volumes) and once with aqueous sodium chloride (10%, 10 volumes). The organic phase was concentrated to 10 volumes, diluted with isopropanol (10 volumes), concentrated to 10 volumes, diluted with isopropanol (10 volumes), stirred at 15°C for 2 hours, filtered, washed and dried to give the crude title compound (-60 g).

[0334] The crude title compound (-50 g) was dissolved in 4:1 THF:MeOH (11 volumes) at 35°C, diluted with methylcyclohexane (0.58 volumes), seeded (2.5 wt%) and stirred for 30 minutes. Methylcyclohexane (12.3 volumes) was added steadily over 10-12 hours. The mixture was stirred at 35°C overnight, filtered, washed with methylcyclohexane (2 volumes), and dried to give the title compound (46.4 g, 84% adjusted yield).

[0335] Alternative Synthesis of Example 6: Batch Chemistry

[0336] 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer

[0337]

[0338] A batch reaction (1 / 6 scale) was performed where a solution of acryloyl chloride (2.19 g, 1.15 eq) in 2-methyltetrahydrofuran (6 volumes) was added dropwise over 30 minutes to a solution of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazepine octatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer (10 g, 21 mmol) in 2-methyltetrahydrofuran (6 volumes) and pH 5.8 buffer [monopotassium phosphate (4.17 g), dipotassium phosphate (0.435 g) and water (to 100 mL volume)]. HPLC analysis after 25 minutes showed 1.63% MA impurity as shown in Table 2.

[0339] Table 2

[0340] Alternative Synthesis of Example 6 %MA Impurity Flow Chemistry 0.33 Batch Chemistry 1.63

[0341] Example 7

[0342] 4-[(4aR)-7-Chloro-9-fluoro-11-oxo-3-prop-2-enoyl-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, P atropisomer

[0343]

[0344] Acryloyl chloride (0.0022 mL, 0.027 mmol) was added to an ice-cooled mixture of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, P-atropisomer (0.013 g, 0.027 mmol) and potassium carbonate (0.011 g, 0.082 mmol) in EtOAc (0.27 mL) and water (0.27 mL). After 20 minutes, the mixture was diluted with EtOAc and brine. The organic extract was dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (20-100% acetone in hexanes) to give the title compound (0.011 g, 78%). ES / MS m / z( 35 Cl / 37 Cl)515 / 517[M+H] + .

[0345] Example 8

[0346] 4-[(4aR)-7-Chloro-9-fluoro-11-oxo-3-prop-2-enoyl-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer

[0347]

[0348] Acryloyl chloride (0.25 M in DCM, 1.1 mL, 0.28 mmol) was added to an ice-cooled mixture of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer (0.128 g, 0.28 mmol) and potassium carbonate (0.12 g, 0.83 mmol) in EtOAc (3 mL) and water (3 mL). After 5 minutes, the layers were diluted with EtOAc and separated. The organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (0-10% 7N ammoniated MeOH in DCM) to give the title compound (0.13 g, 87%). ES / MS m / z( 35 Cl / 37 Cl)515 / 517[M+H] + .

[0349] Bioassay

[0350] The following assays demonstrate that exemplary compounds are inhibitors of KRas G12C and inhibit the growth of certain tumors in vitro and / or in vivo.

[0351] KRas G12C probe occupancy TR-FRET assay

[0352] The purpose of this assay is to measure the ability of the inhibitor to compete with the probe for binding to KRas G12C at codon 12 and covalently modify KRas G12C. Europium on an antibody (Eu-labeled anti-histidine tag antibody LanthaScreen (Eu Anti-His antibody)) bound to KRas G12C is combined with a fluorescent tracer 647 (Alexa Fluor 647) bound to KRas G12C via streptavidin and a biotinylated inhibitor ("KRas probe", see Preparation 223). TM ) generates a signal by time-resolved fluorescence transfer between the

[0353] Inhibitors were tested in a dose response format from 10 mM stocks in 100% DMSO. Labycyte 555 was used for dilution and 100 nL / well containing 10-point 2.8-fold serial dilutions were transferred to the assay plate. Two copies of the assay plate were prepared to measure the potency of the inhibitor after 5 and 60 minutes of incubation with KRas G12C. His-tagged KRas G12C (20 nM) was added to the plate in assay buffer (20 mM Tris-HCl, pH 7.5, 0.01% TX-100 and 1 mM DTT). After 5 or 60 minutes of incubation, 1 μM KRas probe was added and allowed to covalently modify free KRas G12C for 1 hour. This was diluted 4-fold in a buffer containing EuAnti-His antibody and Streptavidin-Coated Tracer 647 (both from Life Technologies) to achieve KRas G12C (5 nM), Anti-Hi antibody (2 nM), KRas probe (300 nM), and Streptavidin-Coated Tracer 647 (500 nM). TM Fluorescence signals were read on a plate reader (excitation at 340 nM, tracer emission (em) at 665 nM, and antibody emission at 615 nM). The maximum control wells lacked inhibitor, while the minimum control wells lacked inhibitor and KRas G12C. The signal ratio (em at 665 / em at 615) was converted to percent inhibition using the following equation: % inhibition = 100 - [(test compound signal - median minimum signal) / (median maximum signal - median minimum signal) x 100]. The fluorescence signals were read on a plate reader using Genedata y = (A + ((BA) / (1 + ((C / x) ^ D)))) The percentage inhibition at each inhibitor concentration was fitted to a four-parameter nonlinear logistic equation to determine the IC 50 , where y = % inhibition, A = minimum asymptote, B = maximum asymptote, and C = relative IC 50 or the inhibitor concentration that produces 50% inhibition within the fitted range of the two asymptotes, and D = Hill slope.

[0354] Compounds within the scope of the present disclosure are evaluated in this assay essentially as described above. Exemplary compounds of the present disclosure evaluated in this assay exhibit KRas G12C inhibitor activity by competing with the probe for binding to KRas G12C at codon 12 and covalently modifying KRAS G12C as shown in Table 3.

[0355] Table 3: KRas G12C probe occupancy TR-FRET assay

[0356] Compound <![CDATA[IC 50 (nM)@5min]]> <![CDATA[IC 50 (nM)@60min]]> Example 1 18600 16000 Example 2 <15 <15 Example 3 451 843 Example 4 <15 <15 Example 5 3270 3250 Example 6 <15 <15 Example 7 26900 17900 Example 8 <15 <15

[0357] As shown in Table 3, Examples 2, 4, 6, and 8 (M atropisomer designations) represent superior IC50 concentrations relative to Examples 1, 3, 5, and 7 (P atropisomer designations).

[0358] Phospho-ERK in H358 cells

[0359] The purpose of this assay is to measure the ability of test compounds to inhibit the phosphorylation of p-ERK1 / 2, a downstream effector of KRas in human lung cancer cells H358 (ATCC CRL-5807). Ultra TM The p-ERK 1 / 2 (Thr202 / Tyr204) assay is a sandwich immunoassay for quantitative detection of phosphorylated-ERK1 / 2 (phosphorylated on Thr202 / Tyr204 in ERK1 or Thr185 / Tyr187 in ERK2) in cell lysates using AlphaTechnology (Perkin Elmer Cat# ALSU-PERK-A50K).

[0360] H358 cells were seeded at 40K cells / well in 100 μL of culture medium (RPMI 1640, GIBCO Cat#22400-071) containing 10% FBS (GIBCO Cat#:10082-147) in a 96-well plate (Costar#3596) and incubated overnight in a humidified chamber at 37°C, 5% CO2. The next morning, 10 μL of serially diluted (3-fold) test compounds (50 μM top concentration) and 10 μL of controls (maximum signal well: 5% DMSO, minimum signal well: 2 μM N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-D]pyrimidin-1(2H)-yl}phenyl)acetamide (trametinib, as a positive control)) were added to the cell plate and incubated for 2 hours at 37°C / 5% CO2 in a humidified tray. Lysis buffer containing a cocktail of protease and phosphatase inhibitors was prepared at ambient temperature. The culture medium was removed by inverting and shaking the cell plate in a sink and then blotting onto a paper towel. Lysis buffer was added to the cell plate (50 μL / well) and the plate was incubated on a shaker at ambient temperature for 10 minutes. For p-ERK detection, acceptor beads were diluted in buffer to a suspension mixture. Using a STARlet liquid handler, 5 μL of acceptor beads and 2 μL of cell lysate were transferred to a 384-well assay plate as a single-step in-tip dilution. The assay plate was sealed with foil and incubated at ambient temperature for 2 hours. Donor beads were diluted in buffer to a suspension mixture. Using a STARlet, 5 μL of donor beads were added to the assay plate, which was then sealed and wrapped with foil. The plate was incubated at ambient temperature in the dark for 2 hours. The plate was then incubated in an EnVision TM Assay plates were read on a plate reader (Perkin Elmer) using the luminescence program.

[0361] Convert the signal to percent inhibition using the following equation:

[0362] % Inhibition = 100 - [(Test compound signal - median minimum signal) / (median maximum signal - median minimum signal) x 100]. The maximum signal is the control well with no inhibitor. The minimum signal is the control well containing the reference inhibitor sufficient to completely inhibit activity. y = (A + ((BA) / (1 + ((C / x) ^ D)))) The percentage inhibition at each inhibitor concentration was fitted to a four-parameter nonlinear logistic equation to determine the IC 50 , where y = % inhibition, A = minimum asymptote, B = maximum asymptote, and C = relative IC 50or the inhibitor concentration that produces 50% inhibition within the fitted range of the two asymptotes, and D = Hill slope.

[0363] Compounds within the scope of the present disclosure were evaluated in this assay essentially as described above. The compounds of the Examples exhibited the ability to inhibit the phosphorylation of p-ERK1 / 2. The data in Table 4 show that the compounds of the Examples exhibited KRas G12C inhibitory activity in this cellular assay.

[0364] Table 4: Phosphorylation-ERK in H358 cells

[0365] Compound <![CDATA[IC 50 (nM)]]> Example 1 2460 Example 2 1.78 Example 3 206 Example 4 1.31 Example 5 330 Example 6 0.649 Example 7 2980 Example 8 0.902

[0366] H358 Cell Active RAS GTPase ELISA

[0367] The purpose of this assay is to measure the ability of the test compound to inhibit constitutive RAS GTPase activity in human lung cancer cells H358 (ATCC CRL-5807). The RAS GTPase ELISA kit (Active Motif Cat#52097) contains a 96-well plate pre-coated with glutathione to capture the GST-Raf-RBD protein supplied with the kit. Activated RAS (GTP-bound) in the cell extract specifically binds to Raf-RBD. The bound RAS is detected with a primary antibody that recognizes human KRas. The secondary antibody conjugated to HRP recognizes the primary antibody, and the developing solution provides a chemiluminescent readout.

[0368] H358 cells were seeded at 80,000 cells / well in 90 μL serum-free medium (RPMI 1640, Gibco) and incubated overnight at 37°C / 5% CO2. The next morning, 10 μL of serially diluted (3-fold) test compounds (500 μM highest concentration) and 10 μL of controls (maximum signal well: 5% DMSO, minimum signal well: 500 μM 1-[4-[6-chloro-8-fluoro-7-(3-hydroxy-1-naphthyl)quinazolin-4-yl]piperazin-1-yl]prop-2-en-1-one, WO2015054572, as inhibitor) were added to the cell plate and incubated at 37°C / 5% CO2 for 2 hours. Complete lysis / binding buffer containing protease inhibitor cocktail and GST-Raf-RBD was prepared and stored on ice. One hour before the cell plate incubation was completed, 50 μL of GST-Raf-RBD was diluted in lysis / binding buffer and the buffer was added to the ELISA assay plate, which was incubated at 4 ° C for 1 hour under gentle shaking. After 2 hours, the cells were washed with 100 μL of ice-cold PBS and lysed with 100 μL of lysis / binding buffer. The cell plate was vibrated for 10 minutes at ambient temperature. The cell plate was then centrifuged at 1500 rpm for 10 minutes at ambient temperature. During this period, 1X washing buffer was prepared at ambient temperature and then used to wash (3x100 μL) GST-Raf-RBD coated assay plates. After washing, 50 μL of cell lysate was added to the GST-Raf-RBD coated assay plate and incubated for 1 hour at ambient temperature under gentle shaking. During this incubation period, 1X antibody binding buffer was prepared and allowed to reach ambient temperature. The assay plate was washed 3x100 μL with 1X Wash Buffer, then 50 μL of primary antibody (diluted 1:500 in 1X Antibody Binding Buffer) was added. The plate was incubated for 1 hour at ambient temperature. The assay plate was washed 3x100 μL with 1X Wash Buffer, then 50 μL of secondary antibody (diluted 1:5000 in 1X Antibody Binding Buffer) was added and incubated for 1 hour at ambient temperature. The assay plate was washed 4x100 μL with 1X Wash Buffer, then 50 μL of Chemiluminescent Working Solution was added at ambient temperature. The plate was then incubated for 1 hour at EnVision TM Assay plates were read on a plate reader (Perkin Elmer) using the luminescence program.

[0369] Convert the signal to percent inhibition using the following equation:

[0370] % Inhibition = 100 - [(Test compound signal - median minimum signal) / (median maximum signal - median minimum signal) x 100]. The maximum signal is the control well with no inhibitor. The minimum signal is the control well containing the reference inhibitor sufficient to completely inhibit activity. y = (A + ((BA) / (1 + ((C / x) ^ D)))) The percentage inhibition at each inhibitor concentration was fitted to a four-parameter nonlinear logistic equation to determine the IC 50 , where y = % inhibition, A = minimum asymptote, B = maximum asymptote, and C = relative IC 50 or the inhibitor concentration that produces 50% inhibition within the fitted range of the two asymptotes, and D = Hill slope.

[0371] Compounds within the scope of the present disclosure were evaluated in this assay essentially as described above. The compounds of the Examples exhibited the ability to inhibit constitutive RAS GTPase activity. The data in Table 5 show that the compounds of the Examples exhibited KRas-GTP inhibitory activity in this human lung cancer cell culture.

[0372] Table 5: H358 cell active RAS GTPase ELISA

[0373] Compound <![CDATA[IC 50 (nM)]]> Example 1 14400 Example 2 6.71 Example 3 902 Example 4 5.66 Example 5 1770 Example 6 3.31 Example 7 23800 Example 8 4.62

[0374] Additional implementation plans:

[0375] Embodiment 1. A compound selected from the group consisting of:

[0376]

[0377]

[0378] or a group consisting of pharmaceutically acceptable salts thereof.

[0379] Embodiment 2. The compound of embodiment 1, which is selected from the following compounds:

[0380]

[0381]

[0382] or a group consisting of pharmaceutically acceptable salts thereof.

[0383] Embodiment 3. The compound of embodiment 1, which is selected from the group consisting of the following compounds:

[0384]

[0385] Embodiment 4. The compound of Embodiment 1, which is selected from the following compounds:

[0386]

[0387]

[0388] or a group consisting of pharmaceutically acceptable salts thereof.

[0389] Embodiment 5. The compound of Embodiment 1, which is selected from the group consisting of:

[0390]

[0391] Embodiment 6. The compound of Embodiment 1, which is selected from the following compounds:

[0392]

[0393] or a group consisting of pharmaceutically acceptable salts thereof.

[0394] Embodiment 7. The compound of Embodiment 1 or 6, wherein the compound is

[0395]

[0396] or a pharmaceutically acceptable salt thereof.

[0397] Embodiment 8. The compound of Embodiment 1 or 6, wherein the compound is

[0398]

[0399] or a pharmaceutically acceptable salt thereof.

[0400] Embodiment 9. The compound of Embodiment 1, which is selected from the following compounds:

[0401]

[0402] or a group consisting of pharmaceutically acceptable salts thereof.

[0403] Embodiment 10. The compound of Embodiment 1 or 9, wherein the compound is

[0404]

[0405] or a pharmaceutically acceptable salt thereof.

[0406] Embodiment 11. The compound of Embodiment 1 or 9, wherein the compound is

[0407]

[0408] or a pharmaceutically acceptable salt thereof.

[0409] Embodiment 12. The compound of Embodiment 1, which is selected from the following compounds:

[0410]

[0411] or a group consisting of pharmaceutically acceptable salts thereof.

[0412] Embodiment 13. The compound of Embodiment 1 or 12, wherein the compound is

[0413]

[0414] or a pharmaceutically acceptable salt thereof.

[0415] Embodiment 14. The compound of Embodiment 1 or 12, wherein the compound is

[0416]

[0417] or a pharmaceutically acceptable salt thereof.

[0418] Embodiment 15. The compound of Embodiment 1, which is selected from the following compounds:

[0419]

[0420] or a group consisting of pharmaceutically acceptable salts thereof.

[0421] Embodiment 16. The compound of Embodiment 1 or 15, wherein the compound is

[0422]

[0423] or a pharmaceutically acceptable salt thereof.

[0424] Embodiment 17. The compound of Embodiment 1 or 15, wherein the compound is

[0425]

[0426] or a pharmaceutically acceptable salt thereof.

[0427] Embodiment 18. A pharmaceutical composition comprising a compound according to any one of Embodiments 1-17 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

[0428] Embodiment 19. A method of treating cancer in a patient, wherein one or more cells express KRas G12C mutant protein, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition according to Embodiment 6, wherein the cancer is selected from lung cancer, advanced non-small cell lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

[0429] Embodiment 20. A method of treating cancer in a patient, wherein one or more cells express KRas G12C mutant protein, comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-17, or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from lung cancer, advanced non-small cell lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

[0430] Embodiment 21. The method according to embodiment 19, wherein the cancer is non-small cell lung cancer.

[0431] Embodiment 22. The method according to embodiment 19 or 20, wherein the cancer is advanced non-small cell lung cancer.

[0432] Embodiment 23. The method according to embodiment 19, wherein the cancer is colorectal cancer.

[0433] Embodiment 24. The method according to embodiment 19, wherein the cancer is pancreatic cancer.

[0434] Embodiment 25. The method of embodiment 19, wherein prior to administering the compound or a pharmaceutically acceptable salt thereof, the patient has a cancer identified as having one or more cells expressing the KRas G12C mutant protein.

[0435] Embodiment 26. A method of treating a patient with cancer having a KRAS G12C mutation, comprising administering to a patient in need thereof an effective amount of a compound according to any one of Embodiments 1-17 or a pharmaceutically acceptable salt thereof.

[0436] Embodiment 27. The method according to any one of Embodiments 19 to 25, wherein the patient is also administered an effective amount of one or more of the following: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

[0437] Embodiment 28. The method according to any one of embodiments 19 to 26, wherein the patient has not received prior therapy with a KRASG12C inhibitor or a pharmaceutically acceptable salt thereof.

[0438] Embodiment 29. A method according to any one of Embodiments 19 to 27, wherein the patient has not received an effective amount of prior therapy with one or more of the following: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

[0439] Embodiment 30. A method according to any one of Embodiments 19 to 27, wherein the patient has received an effective amount of one or more of the following prior therapies: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

[0440] Embodiment 31. The method according to any one of embodiments 19 to 26, wherein the patient has received prior therapy with a KRAS G12C inhibitor or a pharmaceutically acceptable salt thereof.

[0441] Embodiment 32. A method according to any one of Embodiments 19 to 26 or 30, wherein the patient has not received an effective amount of prior therapy with one or more of the following: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

[0442] Embodiment 33. A method according to any one of Embodiments 19 to 26 or 30, wherein the patient has received an effective amount of one or more of the following prior therapies: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

[0443] Embodiment 34. A compound according to any one of Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, for use in therapy.

[0444] Embodiment 35. A compound according to any one of Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

[0445] Embodiment 36. A compound for use according to Embodiment 35, or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from lung cancer, advanced non-small cell lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer and colorectal cancer.

[0446] Embodiment 37. A compound according to any one of Embodiments 1 to 17, or a pharmaceutically acceptable salt thereof, for simultaneous, separate or sequential combination with a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or one or more pharmaceutically acceptable salts thereof for use in treating cancer.

[0447] Embodiment 38. A method for preparing a compound according to Embodiment 1 or 9, comprising:

[0448] The M and P atropisomers of (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester were separated by silica gel flash chromatography.

[0449] Embodiment 39. The method of Embodiment 38, further comprising using 0-30% acetone / hexane.

[0450] Embodiment 40. The method of embodiment 38 or 39, further comprising the steps of:

[0451] TFA was added to a solution of the P-atropisomer of tert-butyl (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate-2-carboxylate in DCM, and then

[0452] Stir at about room temperature for about 3 hours, and

[0453] Concentrate in vacuo.

[0454] Embodiment 41. The method of any one of Embodiments 38 to 40, further comprising:

[0455] The P-atropisomer of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazoctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0456] Embodiment 42. The method of Embodiment 41, further comprising using 4-10% 7N ammoniated MeOH / DCM.

[0457] Embodiment 43. The method of any one of Embodiments 38 to 42, further comprising the steps of:

[0458] A solution of the P atropisomer of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile, atropisomer 1 and DIEA in DCM was cooled to approximately -78°C and then

[0459] Add acryloyl chloride, and after about 5 minutes,

[0460] Dilute with a small amount of 2-propanol, and

[0461] Concentrate in vacuo.

[0462] Embodiment 44. The method of any one of Embodiments 38 to 43, further comprising the steps of:

[0463] The P-atropisomer of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0464] Embodiment 45. The method of any one of Embodiments 38 to 44, further comprising using 20-100% acetone / hexane.

[0465] Embodiment 46. A method for preparing a compound according to Embodiment 1 or 12, comprising:

[0466] (13aS)-tert-butyl 9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylate, tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborinan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate, potassium carbonate, and dichloro[(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) were reacted in 1,4-dioxane in a reaction flask.

[0467] Embodiment 47. The process of Embodiment 46, wherein the 1,4-dioxane has been purged with nitrogen by direct bubbling for 30 minutes.

[0468] Embodiment 48. The method of embodiment 46 or 47, further comprising the steps of:

[0469] The reaction flask was sealed and heated at 105°C for approximately 14 hours and then

[0470] Additional tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate and dichloro[(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) were added, and then

[0471] The reaction flask was sealed and heated at 105°C for approximately 14 hours to produce a reaction mixture.

[0472] Embodiment 49. The method of embodiment 48, further comprising the steps of:

[0473] The reaction mixture was filtered through celite, and

[0474] Rinse with EtOAc to give a filtrate.

[0475] Embodiment 50. The method of embodiment 49, further comprising the steps of:

[0476] The filtrate was concentrated in vacuo, and

[0477] Dilute with EtOAc, water and brine to give an aqueous layer.

[0478] Embodiment 51. The method of any one of Embodiments 46 to 50, further comprising the steps of:

[0479] The aqueous layer was extracted with EtOAc to produce at least one organic extract, then

[0480] The organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo.

[0481] Embodiment 52. The method of any one of Embodiments 46 to 51, further comprising the steps of:

[0482] The M and P atropisomers of (13aS)-tert-butyl 9-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylate were purified by silica gel flash chromatography.

[0483] Embodiment 53. The method of Embodiment 52, further comprising using 10-40% acetone / hexane.

[0484] Embodiment 54. The method of any one of Embodiments 46 to 53, further comprising the steps of:

[0485] TFA was added to a solution of the P-atropisomer of tert-butyl (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate-2-carboxylate in DCM, and

[0486] Stir at about room temperature for about 30 minutes, then

[0487] Concentration in vacuo gave a crude deprotected residue.

[0488] Embodiment 55. The method of embodiment 54, further comprising the steps of:

[0489] The crude deprotected residue was dissolved in DCM and DIEA, then cooled to -78 °C and then

[0490] Acryloyl chloride in DCM was added and after about 30 min concentrated in vacuo.

[0491] Embodiment 56. The method of any one of Embodiments 46 to 55, further comprising the steps of:

[0492] The P-atropisomer of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0493] Embodiment 57. The method of Embodiment 56, further comprising using 20-80% acetone / hexane.

[0494] Embodiment 58. The method of any one of Embodiments 46 to 57, further comprising the steps of:

[0495] HCl gas was bubbled through an ice-cooled solution of the M-atropisomer of tert-butyl (13aS)-9-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate-2-carboxylate in DCM and 2-propanol for approximately 5 minutes.

[0496] Stir at about room temperature for about 5 hours, then

[0497] Cool in an ice bath and then

[0498] HCl gas was bubbled in for about 5 minutes, then

[0499] Stir at about room temperature for about 14 hours, then

[0500] Concentrate in vacuo to give a residue,

[0501] The residue was diluted twice with n-heptane and then

[0502] Concentrate in vacuo and then

[0503] Add MTBE,

[0504] Stir at about room temperature for about 10 minutes, then

[0505] Filtration gave the dihydrochloride salt.

[0506] Embodiment 59. The method of embodiment 58, further comprising the steps of:

[0507] The dihydrochloride salt is dissolved in water, and then

[0508] Add 2-methyltetrahydrofuran and aqueous potassium carbonate solution, then

[0509] Stir vigorously while cooling in an ice bath, then

[0510] Acryloyl chloride in 2-methyltetrahydrofuran was added dropwise and after about 5 minutes,

[0511] Dilute with brine and 2-methyltetrahydrofuran,

[0512] The aqueous layer was extracted twice with 2-methyltetrahydrofuran and then

[0513] The combined organic extracts were washed with water, and

[0514] Dry over magnesium sulfate, filter and concentrate in vacuo.

[0515] Embodiment 60. The method of any one of Embodiments 37 to 54, further comprising the steps of:

[0516] The M atropisomer of 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2-prop-2-enoyl-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepin-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0517] Embodiment 61. The method of Embodiment 60, further comprising using 20-30% 4:1 EtOAc:MeOH / 3:1 EtOAc:hexanes.

[0518] Embodiment 62. A method for preparing a compound according to Embodiment 1 or 15, comprising the steps of:

[0519] (4aR)-7-chloro-9-fluoro-8-iodo-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -3-carboxylic acid tert-butyl ester, N-[3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiophen-2-yl]carbamate and a solution of potassium phosphate in water and 1,4-dioxane were added to the first reaction flask,

[0520] The first reaction flask was purged with nitrogen by direct bubbling for approximately 10 minutes.

[0521] 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride was added to the first reaction flask, and

[0522] The first reaction flask was sealed and heated at about 70°C for about 3 hours to produce a first reaction mixture.

[0523] Embodiment 63. A method for preparing a compound according to any one of Embodiments 1, 15 or 62, comprising the steps of:

[0524] (4aR)-7-chloro-9-fluoro-8-iodo-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine To the second reaction flask, add tert-butyl-3-formate, tert-butyl N-[3-cyano-7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiophen-2-yl]carbamate and a solution of potassium phosphate in water and 1,4-dioxane.

[0525] The second reaction flask was purged with nitrogen by direct bubbling for approximately 10 minutes.

[0526] 1,1'-bis(di-tert-butylphosphino)ferrocenepalladium dichloride was added to the second reaction flask, and

[0527] The second reaction flask was sealed and heated at about 70°C for about 5 hours to produce a second reaction mixture.

[0528] Embodiment 64. The method for preparing a compound according to embodiment 63, further comprising the steps of:

[0529] combining the first and second reaction mixtures to produce a combined reaction mixture,

[0530] The combined reaction mixtures were filtered through celite, and

[0531] Rinse with EtOAc to give a filtrate.

[0532] Embodiment 65. The method for preparing a compound according to embodiment 64, further comprising the steps of:

[0533] The filtrate was diluted with MTBE and saturated aqueous sodium bicarbonate to produce an organic extract, which was then

[0534] Wash the organic extract with brine,

[0535] Dry over magnesium sulfate, filter and concentrate in vacuo to give the crude product.

[0536] Embodiment 66. The method for preparing a compound according to embodiment 65, further comprising the steps of:

[0537] Purification of (4aR)-8-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine by flash chromatography on silica gel -M and P atropisomers of tert-butyl 3-formate.

[0538] Embodiment 67. The method of preparing a compound according to Embodiment 66, further comprising using 0-30% acetone / hexane.

[0539] Embodiment 68. A method for preparing a compound according to Embodiment 66 or 67, further comprising the steps of:

[0540] (4aR)-8-[2-(tert-Butyloxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine was further purified by flash chromatography on silica gel -P-reversion isomer of tert-butyl 3-formate.

[0541] Embodiment 69. The method of preparing a compound according to Embodiment 68, further comprising using 0-100% EtOAc / hexanes.

[0542] Embodiment 70. A method for preparing a compound according to any one of Embodiments 66 to 69, further comprising the steps of:

[0543] TFA was added to (4aR)-8-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -P-atropisomer of tert-butyl 3-carboxylate in DCM, then

[0544] Stir at room temperature for about 30 minutes.

[0545] concentrated in vacuo, and

[0546] The P-atropisomer of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0547] Embodiment 71. A method of preparing a compound according to Embodiment 70, further comprising using 0-10% MeOH / DCM followed by 0-10% 7N ammoniated MeOH / DCM.

[0548] Embodiment 72. A method for preparing a compound according to any one of Embodiments 66 to 71, further comprising the steps of:

[0549] Acryloyl chloride was added to an ice-cooled mixture of the P-atropisomer of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile and potassium carbonate in EtOAc and water. After about 20 minutes,

[0550] Dilute with EtOAc and brine,

[0551] The resulting organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo.

[0552] Embodiment 73. A method for preparing a compound according to any one of embodiments 66 to 72, further comprising:

[0553] The P-atropisomer of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-3-prop-2-enoyl-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0554] Embodiment 74. The method of preparing a compound according to Embodiment 73, further comprising using 20-100% acetone / hexane.

[0555] Embodiment 75. A method for preparing a compound according to any one of Embodiments 66 to 74, further comprising the steps of:

[0556] TFA was added to (4aR)-8-[2-(tert-butoxycarbonylamino)-3-cyano-7-fluoro-benzothiophen-4-yl]-7-chloro-9-fluoro-11-oxo-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepine -M atropisomer of tert-butyl 3-formate in an ice-cooled solution in DCM,

[0557] Stir at room temperature for about 2 hours, and

[0558] Concentrate in vacuo.

[0559] Embodiment 76. A method for preparing a compound according to any one of Embodiments 66 to 75, further comprising the steps of:

[0560] The M atropisomer of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0561] Embodiment 77. A method of preparing a compound according to Embodiment 76, further comprising using 0-10% 7N ammoniated MeOH / DCM.

[0562] Embodiment 78. A method for preparing a compound according to any one of Embodiments 66 to 77, further comprising the steps of:

[0563] Acryloyl chloride was added to an ice-cooled mixture of the M atropisomer of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-1,2,3,4,4a,5-hexahydropyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile and potassium carbonate in EtOAc and water. After about 5 minutes,

[0564] Dilute with EtOAc,

[0565] The organic and aqueous layers were separated, and

[0566] The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo.

[0567] Embodiment 79. A method for preparing a compound according to any one of Embodiments 66 to 78, further comprising the steps of:

[0568] The M atropisomer of 4-[(4aR)-7-chloro-9-fluoro-11-oxo-3-prop-2-enoyl-2,4,4a,5-tetrahydro-1H-pyrazino[2,1-c][1,4]benzoxazepin-8-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile was purified by silica gel flash chromatography.

[0569] Embodiment 80. The method of preparing a compound according to Embodiment 79, further comprising using 0-10% 7N ammoniated MeOH / DCM.

[0570] Embodiment 81. A compound of the formula:

[0571]

[0572] or a pharmaceutically acceptable salt thereof, wherein:

[0573] R is a protecting group.

[0574] Embodiment 82. The compound of Embodiment 81, wherein R is Boc.

[0575] Embodiment 83. The compound of Embodiment 81 or 82, wherein the compound of formula is

[0576]

[0577] or a pharmaceutically acceptable salt thereof.

[0578] Embodiment 84. A compound according to any one of Embodiments 81 to 83, wherein the compound of formula is

[0579]

[0580] Embodiment 85. A compound of the formula:

[0581]

[0582] or a pharmaceutically acceptable salt thereof, wherein:

[0583] R is a protecting group.

[0584] Embodiment 86. The compound of Embodiment 85, wherein R is Boc.

[0585] Embodiment 87. The compound of Embodiment 85 or 86, wherein the compound is

[0586]

[0587] or a pharmaceutically acceptable salt thereof.

[0588] Embodiment 88. The compound of any one of Embodiments 85 to 87, wherein the compound is

[0589]

[0590] Embodiment 89. A method for preparing a compound according to any one of embodiments 81 to 88, comprising:

[0591] Combine (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester with a cyclizing base to give (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester.

[0592] Embodiment 90. A method for preparing a compound according to Embodiment 89, wherein the cyclizing base is selected from the group consisting of sodium hydride, N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), cesium carbonate, diazabicycloundecene (DBU), sodium tert-butoxide, sodium tert-amylate, sodium tert-amylate, potassium tert-amylate and potassium tert-butoxide.

[0593] Embodiment 91. A method for preparing a compound according to embodiment 89 or 90, wherein the cyclizing base is selected from the group consisting of potassium tert-butoxide, sodium tert-pentoxide, potassium tert-pentoxide, sodium tert-butoxide and sodium tert-pentoxide.

[0594] Embodiment 92. The method for preparing a compound according to any one of Embodiments 89 to 91, further comprising using a cyclization solvent.

[0595] Embodiment 93. The method for preparing a compound according to Embodiment 92, wherein the cyclization solvent is selected from the group consisting of DMF, NMP, DMAc, DMSO and THF.

[0596] Embodiment 94. A method for preparing a compound according to any one of embodiments 89 to 93, further comprising:

[0597] Providing sodium tert-amylate in a polar aprotic solvent,

[0598] stirring to form a reactor solution, and

[0599] The reactor solution is cooled to at least about 0°C.

[0600] Embodiment 95. The method for preparing a compound according to Embodiment 94, wherein the polar aprotic solvent is selected from the group consisting of DMAc, NMP, DMSO and DMF.

[0601] Embodiment 96. The method for preparing a compound according to embodiment 94 or 95, wherein the polar aprotic solvent is DMF.

[0602] Embodiment 97. A method of preparing a compound, further comprising:

[0603] (3S)-tert-butyl 4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylate is provided to the reactor solution at about 0°C over at least about 4 hours.

[0604] Embodiment 98. A method for preparing a compound according to any one of Embodiments 94 to 97, further comprising:

[0605] adding a portion of the reactor solution comprising tert-butyl (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylate to water in a range of about 10° C. to about 40° C. over at least about 1 hour to form a preparation mixture,

[0606] The prepared mixture is stirred in the range of about 10°C to about 40°C for at least about 32 hours.

[0607] Embodiment 99. The method for preparing a compound according to Embodiment 98, further comprising:

[0608] separating the solid from the prepared mixture by filtration, and

[0609] The solid from the preparative mixture was washed with a combination of DMF and water.

[0610] Embodiment 100. The method for preparing a compound according to embodiment 98 or 99, further comprising:

[0611] The solid from the preparative mixture was added to EtOH to form a preparative solution.

[0612] Embodiment 101. A method for preparing a compound according to any one of embodiments 98 to 100, further comprising:

[0613] The prepared solution is heated to at least about 30°C and stirred for at least about 15 minutes.

[0614] Embodiment 102. A method for preparing a compound according to any one of Embodiments 98 to 101, further comprising:

[0615] Water is added to the prepared solution over at least about 2 hours.

[0616] Embodiment 103. A method for preparing a compound according to any one of embodiments 98 to 102, further comprising:

[0617] The prepared solution was maintained at about 30°C for at least about 15 minutes.

[0618] Embodiment 104. A method for preparing a compound according to any one of embodiments 98 to 103, further comprising:

[0619] The prepared solution is cooled to at least about 20°C over at least about 2 hours.

[0620] Embodiment 105. The method for preparing a compound according to any one of Embodiments 98 to 104, further comprising:

[0621] The prepared solution is maintained at about 20°C for at least about 2 hours.

[0622] Embodiment 106. A method for preparing a compound according to any one of embodiments 98 to 105, further comprising:

[0623] A solid containing tert-butyl (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate-2-carboxylate was separated, washed and dried from the prepared solution.

[0624] Embodiment 107. A method for preparing a compound according to Embodiment 106, wherein the step of isolating the solid comprising tert-butyl (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate is performed by filtration.

[0625] Embodiment 108. A method for preparing a compound according to embodiment 106 or 107, wherein the step of washing the solid comprising tert-butyl (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate is performed with a combination of EtOH and water.

[0626] Embodiment 109. A method for preparing a compound according to any one of Embodiments 106 to 108, wherein the step of drying the solid comprising tert-butyl (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazoctate is carried out under vacuum at about 50° C. for at least about 16 hours.

[0627] Embodiment 110. A method for preparing a compound according to any one of embodiments 89 to 97, further comprising, before the step of claim 89:

[0628] CDMT was added to the reactor along with acetonitrile and the reactor was cooled to about 0°C, then

[0629] The organic base is added to the reactor and stirred at about 0°C for at least 30 minutes, then

[0630] 4-bromo-2,5-difluorobenzoic acid is added to the reactor,

[0631]

[0632] The mixture was stirred at about 0°C for at least 1 hour to form an acetonitrile coupling solution.

[0633] Embodiment 111. The method for preparing a compound according to embodiment 110, further comprising:

[0634] Add an inorganic base to a separate container, then water, then

[0635] Tert-butyl (3S)-3-(2-hydroxyethyl)piperazine-1-carboxylate:phosphoric acid (1:1) and a base were added to the separate container to form an aqueous solution of piperazine.

[0636] Embodiment 112. The method for preparing a compound according to embodiment 110, wherein the organic base is NMM.

[0637] Embodiment 113. A method for preparing a compound according to Embodiment 111, wherein the inorganic base is K2CO3.

[0638] Embodiment 114. A method for preparing a compound according to any one of embodiments 110 to 113, further comprising:

[0639] The aqueous piperazine solution was added to the acetonitrile coupling solution in the reactor over at least 5 minutes while maintaining an internal temperature of approximately 0°C.

[0640] Embodiment 115. A method for preparing a compound according to any one of embodiments 110 to 114, further comprising:

[0641] The solution was stirred at about 0 °C for at least 2.5 hours.

[0642] Embodiment 116. A method for preparing a compound according to any one of embodiments 110 to 115, further comprising:

[0643] removing a bottom layer of the solution from the reactor,

[0644] The remaining solution was stirred at about 0 °C for at least 64 hours.

[0645] Embodiment 117. A method for preparing a compound according to any one of embodiments 110 to 116, further comprising:

[0646] The remaining solution was concentrated and 2-MeTHF and n-heptane were added.

[0647] Embodiment 118. A method for preparing a compound according to any one of embodiments 110 to 117, further comprising:

[0648] 1 M NaHCO3 aqueous solution was added to the remaining solution and stirred for at least 15 minutes.

[0649] Embodiment 119. A method for preparing a compound according to any one of embodiments 110 to 118, further comprising:

[0650] allowing the layers of the remaining solution to separate,

[0651] removing the bottom layer of the remaining solution from the reactor, and

[0652] The steps of adding 1 M NaHCO 3 aqueous solution, stirring and removing the bottom layer were repeated.

[0653] Embodiment 120. A method for preparing a compound according to any one of embodiments 110 to 119, further comprising:

[0654] 0.05 M aqueous HCl was added to the remaining solution and stirred for at least 15 minutes.

[0655] Embodiment 121. A method for preparing a compound according to any one of embodiments 110 to 120, further comprising:

[0656] allowing the layers of the remaining solution to separate,

[0657] The bottom layer of the remaining solution is removed from the reactor.

[0658] Embodiment 122. A method for preparing a compound according to any one of embodiments 110 to 121, further comprising:

[0659] Water was added to the remaining solution and stirred for at least 15 minutes.

[0660] Embodiment 123. A method for preparing a compound according to any one of embodiments 110 to 122, further comprising:

[0661] allowing the layers of the remaining solution to separate,

[0662] The bottom layer of the remaining solution is removed from the reactor.

[0663] Embodiment 124. A method for preparing a compound according to any one of embodiments 110 to 123, further comprising:

[0664] The remaining solution was concentrated and 2-MeTHF was added, and

[0665] The concentration and 2-MeTHF addition steps were repeated.

[0666] Embodiment 125. A method for preparing a compound according to any one of embodiments 110 to 124, further comprising:

[0667] The remaining solution was concentrated and DMF was added to afford (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester.

Claims

1. A compound of the formula, or a pharmaceutically acceptable salt thereof, It can be obtained by combining (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylic acid tert-butyl ester, N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamic acid tert-butyl ester, potassium carbonate and a catalyst in 1,4-dioxane, wherein the M:P atropisomer ratio is at least 4:

1.

2. The compound according to claim 1, which is further obtainable by adding additional tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate and the catalyst.

3. The compound according to claim 1 or 2, wherein the catalyst comprises diacetate [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl] palladium(II), dichloro [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl] palladium(II), dichloro [(R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl] palladium(II), dichloro 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloromethane, dichloro (1,2-bis(diphenylphosphino)ethane) palladium(II), dichloro (1,2-bis(diphenylphosphino)ethane) palladium(II), dichloro (1,1'-bis(diphenylphosphino)ethane) palladium(II). 3-bis(diphenylphosphino)propane)palladium(II), dichloro[1,4-bis(diphenylphosphino)butane]palladium(II), dichloro[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene]palladium(II), dibromo[1,1'-bis(diphenylphosphino)ferrocene]palladium(II), [(1,2,3-η)-2-buten-1-yl]chloro[dicyclohexyl(2',6'-dimethoxy[1,1'-biphenyl]-2-yl)phosphine-κP]palladium, [(1,2,3-η)-2-buten-1-yl]chloro(tricyclohexylphosphine)palladium or trans-dichlorobis(tricyclohexylphosphine)palladium(II).

4. The compound according to any one of claims 1 to 3, wherein the catalyst is [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) diacetate or [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) dichloride.

5. A compound of the formula or a pharmaceutically acceptable salt thereof, It can be obtained by combining acryloyl chloride and 4-[(13aS)-10-chloro-8-fluoro-6-oxo-2,3,4,12,13,13a-hexahydro-1H-pyrazino[2,1-d][1,5]benzoxazacyclooctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M atropisomer, in a continuous stirred tank reactor. wherein the Michael adduct impurity, 4-[(13aS)-2-[3-[(13aS)-9-(2-amino-3-cyano-7-fluoro-benzothiophen-4-yl)-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazaoctatetraen-2-yl]-3-oxo-propyl]-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazaoctatetraen-9-yl]-2-amino-7-fluoro-benzothiophene-3-carbonitrile, M,M atropisomers are less than 1% as measured by HPLC analysis 6. A compound of the formula or a pharmaceutically acceptable salt thereof.

7. The compound according to claim 6, wherein the compound is 8. A compound of the formula or a pharmaceutically acceptable salt thereof, It can be obtained by combining (3S)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester:phosphoric acid (1:1), base and 4-bromo-2,5-difluorobenzoic acid to give (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester.

9. A compound of the formula or a pharmaceutically acceptable salt thereof.

10. The compound according to claim 9, wherein the compound is 11. A compound of the formula or a pharmaceutically acceptable salt thereof, It can be obtained by combining (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester with a cyclizing base to give (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester.

12. The compound of claim 11, wherein the cyclizing base comprises sodium hydride, N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), cesium carbonate, diazabicycloundecene (DBU), sodium tert-butoxide, sodium tert-amylate, sodium tert-amylate, potassium tert-amylate, or potassium tert-butoxide.

13. The compound according to claim 11 or 12, wherein the cyclizing base comprises potassium tert-butoxide, sodium tert-pentoxide, potassium tert-amylate, sodium tert-butoxide or sodium tert-amylate.

14. The compound according to any one of claims 11 to 13, which is obtainable by further comprising a cyclization solvent.

15. The compound of claim 14, wherein the cyclization solvent comprises DMF, NMP, DMAc, DMSO or THF.

16. A compound of the formula or a pharmaceutically acceptable salt thereof, It can be obtained by combining (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepinatetraene-2-carboxylic acid tert-butyl ester with a chlorinating agent to give (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepinatetraene-2-carboxylic acid tert-butyl ester.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

18. A method for treating cancer in a patient, wherein one or more cells express KRas G12C mutant protein, comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition of claim 17, wherein the cancer is selected from lung cancer, advanced non-small cell lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

19. A method for treating cancer in a patient, wherein one or more cells express KRas G12C mutant protein, comprising administering to a patient in need thereof an effective amount of a compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from lung cancer, advanced non-small cell lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer, and colorectal cancer.

20. The method of claim 19, wherein the cancer is non-small cell lung cancer.

21. The method of claim 19, wherein the cancer is advanced non-small cell lung cancer.

22. The method of claim 19, wherein the cancer is colorectal cancer.

23. The method of claim 19, wherein the cancer is pancreatic cancer.

24. The method of claim 19, wherein prior to administering the compound or a pharmaceutically acceptable salt thereof, the patient has a cancer identified as having one or more cells expressing the KRas G12C mutant protein.

25. A method for treating a patient with cancer having a KRAS G12C mutation, comprising administering to a patient in need thereof an effective amount of a compound according to any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof.

26. The method according to any one of claims 18 to 25, wherein an effective amount of one or more of the following is also administered to the patient: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

27. The method of any one of claims 18 to 26, wherein the patient has not received prior therapy with a KRAS G12C inhibitor or a pharmaceutically acceptable salt thereof.

28. The method of any one of claims 18 to 27, wherein the patient has not received an effective amount of prior therapy with one or more of the following: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

29. The method according to any one of claims 18 to 27, wherein the patient has received an effective amount of one or more of the following prior therapies: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

30. The method of any one of claims 18 to 26, wherein the patient has received prior therapy with a KRAS G12C inhibitor or a pharmaceutically acceptable salt thereof.

31. The method of any one of claims 18 to 26 or 30, wherein the patient has not received an effective amount of prior therapy with one or more of the following: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

32. The method of any one of claims 18 to 26 or 30, wherein the patient has received an effective amount of one or more of the following prior therapies: a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, a SHP2 inhibitor or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof.

33. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for use in therapy.

34. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

35. The compound for use according to claim 34, or a pharmaceutically acceptable salt thereof, wherein the cancer is selected from lung cancer, advanced non-small cell lung cancer, pancreatic cancer, cervical cancer, esophageal cancer, endometrial cancer, ovarian cancer, bile duct cancer and colorectal cancer.

36. A compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, for use in simultaneous, separate or sequential combination with one or more of a PD-1 inhibitor, a PD-L1 inhibitor, a CD4 / CDK6 inhibitor or a pharmaceutically acceptable salt thereof, an EGFR inhibitor or a pharmaceutically acceptable salt thereof, an ERK inhibitor or a pharmaceutically acceptable salt thereof, a platinum agent, pemetrexed, an Aurora A inhibitor or a pharmaceutically acceptable salt thereof, or a SHP2 inhibitor or a pharmaceutically acceptable salt thereof for treating cancer.

37. A method for preparing the compound according to claim 6, the method comprising: Combine (3S)-tert-butyl 3-(2-hydroxyethyl)piperazine-1-carboxylate:phosphoric acid (1:1), base and 4-bromo-2,5-difluorobenzoic acid to give (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylate.

38. A method for preparing the compound according to claim 9, the method comprising: Combine (3S)-4-(4-bromo-2,5-difluoro-benzoyl)-3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester with a cyclizing base to give (13aS)-9-bromo-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester.

39. The method of claim 38, wherein the cyclizing base comprises sodium hydride, N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), cesium carbonate, diazabicycloundecene (DBU), sodium tert-butoxide, sodium tert-amylate, sodium tert-amylate, potassium tert-amylate, or potassium tert-butoxide.

40. The method of claim 38 or 39, wherein the cyclizing base comprises potassium tert-butoxide, sodium tert-pentoxide, potassium tert-amylate, sodium tert-butoxide, or sodium tert-amylate.

41. The method of any one of claims 38 to 40, wherein the method further comprises a cyclization solvent.

42. The method of claim 41, wherein the cyclization solvent comprises DMF, NMP, DMAc, DMSO, or THF.

43. A method for preparing a compound of the formula, or a pharmaceutically acceptable salt thereof, The method comprises: (13aS)-9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazepine-2-carboxylic acid tert-butyl ester, N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamic acid tert-butyl ester, potassium carbonate and dichloro[(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) are reacted in a solvent.

44. A method for preparing a compound of the formula, or a pharmaceutically acceptable salt thereof, The method comprises: (13aS)-tert-butyl 9-bromo-10-chloro-8-fluoro-6-oxo-1,3,4,12,13,13a-hexahydropyrazino[2,1-d][1,5]benzoxazacyclooctatetraene-2-carboxylate, [(S)-(-)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl]palladium(II) diacetate, tert-butyl N-[3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborolan-2-yl)-7-fluoro-benzothiophen-2-yl]carbamate, a base and 1,1,1-tris(hydroxymethyl)ethane are reacted.

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