Improved synthesis of key intermediates of KRAS G12C inhibitor compounds

Abstract

The present invention relates to improved synthesis of key intermediates of KRAS G12C inhibitor compounds. The present invention relates to an improved, efficient, scalable process to prepare intermediate compounds of structure (I), e.g., Compound 5M, which can be used to synthesize compounds targeting KRAS G12C mutations, e.g., (II).

Description

[0001] This application is a divisional application of an invention patent with an application date of November 15, 2019, application number 201980075080.8, and invention name “Improved Synthesis of Key Intermediates of KRASG12C Inhibitor Compounds”. Technical Field

[0002] The present invention relates to an improved, efficient and scalable method for preparing structural The intermediate compounds, such as compound 5M, can be used to synthesize compounds that inhibit KRAS G12C mutation. Background Art

[0003] KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, thyroid cancer, and bile duct cancer. KRAS mutations are also observed in approximately 25% of NSCLC patients, and some studies have indicated that KRAS mutations are negative prognostic factors in NSCLC patients. Recently, V-Ki-ras2Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations have been found to confer resistance to epidermal growth factor receptor (EGFR) targeted therapy in colorectal cancer; therefore, the mutation status of KRAS can provide important information before prescribing TKI therapy. Overall, new medical treatments are needed for patients with pancreatic cancer, lung adenocarcinoma, or colorectal cancer, especially those who have been diagnosed with such cancers characterized by KRAS mutations and including those who have progressed after chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0004] Figure 1 The crystal arrangement of composition 4a is shown.

[0005] Figure 2-1 An XRPD overlay of the diketo racemates Forms A to E is shown.

[0006] Figure 2-2 An XRPD overlay of the (1S)-(-)-camphoric acid cocrystal is shown.

[0007] Figure 2-3 An XRPD overlay of (+)-2,3-dibenzoyl-D-tartaric acid cocrystals is shown.

[0008] Figure 2-4 An XRPD overlay of D-(+)-malic acid cocrystals is shown.

[0009] Figure 2-5 Shown are XRPD overlays of meta-diketone co-crystals at different temperatures.

[0010] Figure 2-6 Shown are overlays of XRPD patterns for the diketone co-crystals at different temperatures.

[0011] Figure 2-7 Shown are XRPD overlays of the meta-diketone co-crystal and para-diketone co-crystal mixtures at different temperatures.

[0012] Figure 2-8 Shown are overlays of XRPD patterns of the diketone racemate at different temperatures (I / II).

[0013] Figure 2-9 Shown are overlays of XRPD patterns of the diketone racemate at different temperatures (II / II).

[0014] Figure 2-10 The ternary phase diagram of the meta / para-diketone eutectic is shown.

[0015] Figure 2-11 The ternary phase diagram for meta / para-diketones is shown.

[0016] Figure 3-1 The XRPD of the diketone racemate Form A is shown.

[0017] Figure 3-2 A TGA / DSC overlay of the diketone racemate Form A is shown.

[0018] Figure 3-3 The A-type diketone racemate is shown 1 H NMR spectrum.

[0019] Figure 3-4 A PLM image of the diketone racemate Form A is shown.

[0020] Figure 3-5 The XRPD of the Form B diketone racemate is shown.

[0021] Figure 3-6 A TGA / DSC overlay of the Form B diketone racemate is shown.

[0022] Figure 3-7 The B-type diketone racemate is shown 1 H NMR spectrum.

[0023] Figure 3-8 The XRPD of the Form C diketone racemate is shown.

[0024] Figure 3-9 A TGA / DSC overlay of the diketone racemate Form C is shown.

[0025] Figure 3-10 The C-type diketone racemate is shown 1 H NMR spectrum.

[0026] Figure 3-11 The XRPD of the D-form diketone racemate is shown.

[0027] Figure 3-12 Shown is a TGA / DSC overlay of the D-dione racemate.

[0028] Figure 3-13 The D-type diketone racemate is shown 1 H NMR spectrum.

[0029] Figure 3-14 The XRPD of the diketone racemate Form E is shown.

[0030] Figure 3-15 Shown is a TGA / DSC overlay of the E-type diketone racemate.

[0031] Figure 3-16 The E-type diketone racemate is shown 1 H NMR spectrum.

[0032] Figure 3-17 The XRPD of Form A meta-diketone cocrystal is shown.

[0033] Figure 3-18 Shown is a TGA / DSC overlay of Form A meta-diketone cocrystal.

[0034] Figure 3-19 The A-type meta-diketone cocrystal is shown 1 H NMR spectrum.

[0035] Figure 3-20 XRPD of Form A para-diketone cocrystal is shown.

[0036] Figure 3-21 Shown is a TGA / DSC overlay of Form A para-diketone cocrystal.

[0037] Figure 3-22 The A-type diketone cocrystal is shown 1 H NMR spectrum.

[0038] Figure 3-23 An XRPD overlay of the diketone racemic form is shown.

[0039] Figure 3-24 XRPD of a competitive slurry sample is shown.

[0040] Figure 3-25 Shown is an XRPD overlay of the prepared p-diketone co-crystals.

[0041] Figure 3-26 shows the meta / para-diketone cocrystal 1 Overlay of H NMR spectra.

[0042] Figure 3-27Shown is an XRPD overlay of the prepared p-diketone co-crystals.

[0043] Figure 4-1 Interconversion diagrams of meta-diketone DBTA cocrystal forms are shown.

[0044] Figure 5-1 An XRPD overlay of meta-diketone DBTA co-crystal forms (Forms A to E) is shown.

[0045] Figure 5-2 XRPD overlays of meta-diketone DBTA co-crystal forms (Forms F-K) are shown.

[0046] Figure 5-3 An XRPD overlay of the meta-diketone DBTA co-crystal forms (Form L-Q) is shown.

[0047] Figure 5-4 The XRPD pattern of Form A is shown.

[0048] Figure 5-5 The TGA / DSC curve of Form A is shown.

[0049] Figure 5-6 Shows type A 1 H NMR.

[0050] Figure 5-7 An XRPD overlay of Form B is shown.

[0051] Figure 5-8 The TGA / DSC curve of Form B is shown.

[0052] Figure 5-9 Shows type B 1 H NMR.

[0053] Figure 5-10 The XRPD pattern of Form C is shown.

[0054] Figure 5-11 The TGA / DSC curve of Form C is shown.

[0055] Figure 5-12 Shows C-type 1 H NMR.

[0056] Figure 5-13 The XRPD pattern of Form D is shown.

[0057] Figure 5-14 The TGA / DSC curve of Form D is shown.

[0058] Figure 5-15 Shows the D-type 1 H NMR.

[0059] Figure 5-16 The XRPD pattern of Form E is shown. Figure 5-17 The TGA / DSC curve of Form E is shown. Figure 5-18 Shows the E-type 1 H NMR.

[0060] Figure 5-19 The XRPD pattern of Form F is shown. Figure 5-20 The TGA / DSC curve of Form F is shown. Figure 5-21 Shows the F type 1 H NMR.

[0061] Figure 5-22 The XRPD pattern of Form G is shown. Figure 5-23 The TGA / DSC curve of Form G is shown.

[0062] Figure 5-24 Shows the G type 1 H NMR.

[0063] Figure 5-25 The XRPD pattern of Form H is shown. Figure 5-26 The TGA / DSC curve of Form H is shown.

[0064] Figure 5-27 Shows H-type 1 H NMR.

[0065] Figure 5-28 The XRPD pattern of Form I is shown.

[0066] Figure 5-29 The TGA / DSC curve of Form I is shown.

[0067] Figure 5-30 Shows type I 1 H NMR.

[0068] Figure 5-31 The XRPD pattern of Form J is shown.

[0069] Figure 5-32 A J-shaped TGA / DSC curve is shown.

[0070] Figure 5-33 Shows a J-shaped 1 H NMR.

[0071] Figure 5-34 The XRPD pattern of Form K is shown. Figure 5-35 The TGA / DSC curve of Form K is shown.

[0072] Figure 5-36 Shows K-type 1 H NMR.

[0073] Figure 5-37 The XRPD pattern of Form L is shown. Figure 5-38 The TGA / DSC curve of Form L is shown.

[0074] Figure 5-39 Shows L-shaped 1 H NMR.

[0075] Figure 5-40 The XRPD pattern of Form M is shown. Figure 5-41 The TGA / DSC curve of Form M is shown.

[0076] Figure 5-42 Shows M-type 1 H NMR.

[0077] Figure 5-43 The XRPD pattern of Form N is shown.

[0078] Figure 5-44 The TGA / DSC curve of Form N is shown.

[0079] Figure 5-45 Shows N-type 1 H NMR.

[0080] Figure 5-46 The XRPD pattern of Form O is shown.

[0081] Figure 5-47 The TGA / DSC curve of Form O is shown.

[0082] Figure 5-48 Shows O-type 1 H NMR.

[0083] Figure 5-49 The XRPD pattern of Form P is shown.

[0084] Figure 5-50 The TGA / DSC curve of Form P is shown.

[0085] Figure 5-51 Shows P-type 1 H NMR.

[0086] Figure 5-52 The XRPD pattern of Form Q is shown.

[0087] Figure 5-53 The TGA / DSC curve of form Q is shown.

[0088] Figure 5-54 Shows Q-type 1 H NMR.

[0089] Figure 6-1The HPLC of M-5 resolved with 1,3-diphenyl-3-oxopropanesulfonic acid is shown.

[0090] Figure 6-2 HPLC of 5 (p-atropisomer excess) resolved with 1,3-diphenyl-3-oxopropanesulfonic acid is shown. Summary of the invention

[0091] The present invention relates to the improved preparation of compounds having the following chemical structure:

[0092]

[0093] and its key intermediates, i.e., compositions and compounds containing the following chemical structures:

[0094]

[0095] The present invention also relates to a method for preparing compound 5M having the following chemical structure:

[0096]

[0097] The present invention also relates to a composition comprising the following structure:

[0098] DETAILED DESCRIPTION

[0099] definition

[0100] Abbreviations: The following abbreviations may be used in this document:

[0101]

[0102]

[0103]

[0104] Unless otherwise indicated, the use of the terms "a or an" and "the" and similar references in the context of describing the present invention (especially in the context of the scope of the claims) should be interpreted to cover both the singular and the plural. Unless otherwise indicated herein, the statement of the range of values ​​herein is intended only to be used as a shorthand method of referring individually to each independent value within the range, and each independent value is incorporated into the specification as if the value were individually stated herein. Unless otherwise required, any and all examples provided herein, or the use of exemplary language (e.g., "for example") are intended to better illustrate the present invention and are not intended to limit the scope of the present invention. No language in this specification should be interpreted as indicating that any non-required element is essential to practicing the present invention.

[0105] As used herein, the term "alkyl" refers to a straight or branched C1-C8 hydrocarbon group, including but not limited to methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl and 2-ethylbutyl. The term C m-n The term "alkylene" refers to an alkyl group having "m" to "n" carbon atoms. The term "alkylene" refers to an alkyl group having a substituent. An alkyl (e.g., methyl) or alkylene (e.g., -CH2-) group may be substituted with one or more, and typically one to three, groups independently selected from, for example, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, -NC, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5-C 10 Aryl and C5-C 10 Heteroaryl. The term "haloalkyl" specifically refers to an alkyl group wherein at least one (eg, 1 to 6) or all of the hydrogens of the alkyl group are replaced by halogen atoms.

[0106] The terms "alkenyl" and "alkynyl" refer to an alkyl group further comprising a double bond or a triple bond, respectively.

[0107] As used herein, the term "halo" refers to fluoro, chloro, bromo, and iodo. The term "alkoxy" is defined as -OR, where R is alkyl.

[0108] As used herein, the term "amino" or "amine" refers interchangeably to a -NR2 group, wherein each R is, for example, H or a substituent. In some embodiments, the amino group is further substituted to form an ammonium ion, for example, NR3 + . The ammonium moiety is specifically included in the definition of "amino" or "amine". The substituent may be, for example, an alkyl, alkoxy, cycloalkyl, heterocycloalkyl, amide or formate. The R group may be further substituted, for example, with one or more (e.g., one to four) groups selected from the group consisting of halide, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, formate, amine and amide. An "amide" or "amido" group interchangeably refers to a group similar to an amine or amino group but additionally includes C(O), for example -C(O)NR2.

[0109] As used herein, the term "aryl" refers to a C 6-14A monocyclic or polycyclic aromatic group, preferably C 6-10 A monocyclic or bicyclic aromatic group, or C 10-14 Polycyclic aromatic groups. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthracenyl, phenanthrenyl, pyrenyl, biphenyl and terphenyl. Aryl also refers to C 10-14 Bicyclic and tricyclic carbocycles in which one ring is aromatic and the other ring is saturated, partially unsaturated or aromatic, for example dihydronaphthyl, indenyl, indanyl or tetrahydronaphthyl (tetrahydronaphthyl). Unless otherwise indicated, aryl groups may be unsubstituted or substituted by one or more and in particular one to four groups independently selected from, for example, halo, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, -CF3, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5-C 10 Aryl and C5-C 10 Heteroaryl.

[0110] As used herein, the term "cycloalkyl" refers to a monocyclic or polycyclic non-aromatic carbocyclic ring, wherein the polycyclic rings may be fused, bridged or spirocyclic. The carbocyclic ring may have 3 to 10 carbon ring atoms. Contemplated carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl.

[0111] As used herein, the term "heterocycloalkyl" means a monocyclic or polycyclic (e.g., bicyclic), saturated or partially unsaturated ring system containing 3 or more (e.g., 3 to 12, 4 to 10, 4 to 8, or 5 to 7) total atoms, of which 1-5 (e.g., 1, 2, 3, 4, or 5) atoms are independently selected from nitrogen, oxygen, and sulfur. Non-limiting examples of heterocycloalkyl include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, dihydropyrrolyl, morpholinyl, thiomorpholinyl, dihydropyridinyl, oxepinyl, dioxepinyl, thioepinyl, and diazepanyl.

[0112] Unless otherwise indicated, a cycloalkyl or heterocycloalkyl group may be unsubstituted or substituted with one or more and specifically one to four groups. Some contemplated substituents include halo, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10Cycloalkyl, C3-C 10 Heterocycloalkyl, C5-C 10 Aryl and C5-C 10 Heteroaryl.

[0113] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic ring system (e.g., bicyclic) containing one to three aromatic rings and one to four (e.g., 1, 2, 3, or 4) heteroatoms selected from nitrogen, oxygen, and sulfur in the aromatic rings. In certain embodiments, heteroaryl has 5 to 20, 5 to 15, 5 to 10 rings, or 5 to 7 atoms. Heteroaryl also refers to C 10-14 Bicyclic and tricyclic rings, one of which is aromatic and the other rings are saturated, partially unsaturated or aromatic. Examples of heteroaryl include, but are not limited to, furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl, benzofuranyl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzophenylthio, benzothiophene, benzothiophenyl ... The invention also includes the following: and triazolyl, benzoxazolyl, furopyridinyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothiophenyl, isoindolyl, isoquinolyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridinyl, pyrrolopyridinyl, quinolyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidinyl and thienopyridinyl. Unless otherwise indicated, the heteroaryl group may be unsubstituted or substituted with one or more and particularly one to four or one or two substituents. Contemplated substituents include halo, C 1-8 Alkyl, C 2-8 Alkenyl, C 2-8 Alkynyl, -OCF3, -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO2C1-C8 alkyl, -OCOC1-C8 alkyl, C3-C 10 Cycloalkyl, C3-C 10 Heterocycloalkyl, C5-C 10 Aryl and C5-C 10 Heteroaryl.

[0114] As used herein, the term Boc refers to the structure

[0115] Example

[0116] Example 1

[0117] In one embodiment of the present invention, the present invention includes a composition comprising a compound having Formula 4:

[0118]

[0119] and a compound having formula B:

[0120]

[0121] Example 2

[0122] In another embodiment of the present invention, the present invention includes the composition as described in Example 1, wherein the compound having Formula 4 is a compound having Formula 5M:

[0123]

[0124] Example 3

[0125] In another embodiment of the present invention, the present invention includes the composition as described in Example 1, wherein the compound having Formula 4 is a compound having Formula 5P:

[0126]

[0127] Example 4

[0128] In another embodiment of the present invention, the present invention includes a composition as described in any one of embodiments 1-3, wherein the compound having formula B is a compound having formula B1:

[0129]

[0130] Example 5

[0131] In another embodiment of the present invention, the present invention includes a composition as described in any one of embodiments 1-3, wherein the compound having formula B is a compound having formula B2:

[0132]

[0133] Example 6

[0134] In another embodiment of the present invention, the present invention includes the composition of any one of embodiments 1-5, wherein the composition comprises a compound having formula 4 and a compound having formula B in a 2:1 ratio.

[0135] Example 7

[0136] In another embodiment of the present invention, the present invention includes the composition of any one of embodiments 1-6, wherein the composition further comprises 2-methyltetrahydrofuran having the formula:

[0137]

[0138] Example 8

[0139] In another embodiment of the present invention, the present invention includes the composition of any one of embodiments 1-7, wherein the ratio of 2-methyltetrahydrofuran to the compound having formula B is 2:1.

[0140] Example 9

[0141] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 1, wherein the composition has the following formula:

[0142]

[0143] Example 10

[0144] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 9, wherein the composition has the following formula:

[0145]

[0146] Embodiment 11

[0147] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 9, wherein the composition has the following formula:

[0148]

[0149] Example 12

[0150] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 9, wherein the composition has the following formula:

[0151]

[0152] Example 13

[0153] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 9, wherein the composition has the following formula:

[0154]

[0155] Embodiment 14

[0156] In another embodiment of the present invention, the present invention includes the composition of any one of embodiments 1-13, wherein the composition is in a crystalline state.

[0157] Embodiment 15

[0158] In another embodiment of the present invention, the present invention includes a method for preparing a composition having formula 4a, the method comprising reacting a compound 4 having the following chemical structure in the presence of 2-methyltetrahydrofuran:

[0159] 4 reacts with a compound B1 having the following formula

[0160]

[0161] To form a composition of Formula 4a having the following structure:

[0162] Example 16

[0163] In another embodiment of the present invention, the present invention includes a method of obtaining a compound of formula 5M having the following chemical structure:

[0164]

[0165] The method comprises:

[0166] a) in the presence of 2-methyltetrahydrofuran, reacting a compound 4 having the following chemical structure:

[0167] 4 reacts with a compound B1 having the following formula

[0168]

[0169] To form a composition of formula 4a having the following structure in crystalline form:

[0170]

[0171] b) isolating composition 4a, and

[0172] c) treating said isolated composition 4a with a base to produce said compound of formula 5M.

[0173] Embodiment 17

[0174] In another embodiment of the present invention, the present invention comprises the method as described in embodiment 16, wherein the base is Na2HPO4.

[0175] Embodiment 18

[0176] In another embodiment of the present invention, the present invention comprises the method as described in embodiment 16, wherein the base is NaHCO3.

[0177] Embodiment 19

[0178] In another embodiment of the present invention, the present invention includes a composition comprising a compound having Formula 4:

[0179]

[0180] and a compound having formula 11:

[0181]

[0182] Embodiment 20

[0183] In another embodiment of the present invention, the present invention includes the composition as described in Example 19, wherein the compound having Formula 4 is a compound having Formula 5M:

[0184]

[0185] Embodiment 21

[0186] In another embodiment of the present invention, the present invention includes the composition as described in Example 19, wherein the compound having Formula 4 is a compound having Formula 5P:

[0187]

[0188] Embodiment 22

[0189] In another embodiment of the present invention, the present invention includes a composition as described in any one of embodiments 19-21, wherein the compound having formula 11 is a compound having formula 11a:

[0190]

[0191] Embodiment 23

[0192] In another embodiment of the present invention, the present invention includes a composition as described in any one of embodiments 19-21, wherein the compound having formula 11 is a compound having formula 11b

[0193]

[0194] Embodiment 24

[0195] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 19, wherein the composition has the following formula:

[0196]

[0197] Embodiment 25

[0198] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 19, wherein the composition has the following formula:

[0199]

[0200] Embodiment 26

[0201] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 19, wherein the composition has the following formula:

[0202]

[0203] Embodiment 27

[0204] In another embodiment of the present invention, the present invention includes the composition as described in embodiment 19, wherein the composition has the following formula:

[0205]

[0206] Embodiment 28

[0207] In another embodiment of the present invention, the present invention includes a composition as described in any one of embodiments 19-27, wherein the composition comprises a compound having formula 4 and a compound having formula 11 in a 1:1 ratio.

[0208] Embodiment 29

[0209] In another embodiment of the present invention, the present invention includes the method as described in Example 16, wherein the compound having formula 5M is used to produce a compound having the following formula 9:

[0210] Embodiment 30

[0211] The method of embodiment 29, wherein the method further comprises mixing the compound of formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

[0212] Compounds of the present disclosure

[0213] Provided herein are KRAS inhibitors having structures discussed in more detail below.

[0214] The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds, wherein one or more atoms of the compounds disclosed herein are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, for example, 2 H. 3 H. 11 C. 13 C.14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. These radiolabeled compounds can be used to help determine or measure the effectiveness of a compound by characterizing, for example, the site or mode of action, or binding affinity to a pharmacologically important site of action. Certain isotope-labeled compounds of the present disclosure (e.g., those incorporating a radioisotope) are useful in drug and / or substrate tissue distribution studies. Given the ease of incorporation and the ready-to-use detection format, the radioisotope tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) is particularly useful for this purpose.

[0215] Using heavier isotopes (such as deuterium, 2 H) substitutions may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and are therefore preferred in some circumstances.

[0216] Using positron-emitting isotopes (e.g. 11 C. 18 F. 15 O and 13 N) substitution can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art, or by methods analogous to those described in the Preparations and Examples described below, using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously employed.

[0217] Isotopically-labeled compounds as disclosed herein can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Schemes, using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

[0218] Certain compounds as disclosed herein can exist as stereoisomers (i.e., isomers with different only spatial arrangements of atoms), including optical isomers and conformers (or conformers). Compounds disclosed herein include all stereoisomers as pure individual stereoisomer preparations and enriched preparations of each, as well as racemic mixtures of such stereoisomers, and individual diastereomers and enantiomers that can be separated according to methods known to those skilled in the art. In addition, compounds disclosed herein include all tautomeric forms of these compounds.

[0219] Certain compounds disclosed herein may exist as atropisomers, which are conformational stereoisomers that occur when rotation about a single bond in a molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers as pure preparations of individual atropisomers, enriched preparations of each, or non-specific mixtures of each. If the barrier to rotation about a single bond is high enough and the interconversion between conformations is slow enough, separation and isolation of the isomeric species may be tolerated. For example, groups such as, but not limited to, the following groups and May exhibit restricted rotation.

[0220] The term "monohydrate" refers to a salt of Compound 9 that has about one associated water molecule. Those skilled in the art will appreciate that the exact number of associated water molecules may vary slightly at any time with variable temperature, pressure, and other environmental influences. All slight variations in the number of associated water molecules are contemplated within the scope of the present invention.

[0221] The term "dihydrate" refers to a salt of compound 9 that has about two associated water molecules. One skilled in the art will appreciate that the exact number of associated water molecules may vary slightly at any time with variable temperature, pressure, and other environmental influences. All slight variations in the number of associated water molecules are contemplated within the scope of the present invention.

[0222] The term "co-crystal" refers to a crystalline material comprising two or more compounds at ambient temperature (20°C to 25°C, preferably 20°C), at least two of which are held together by weak interactions, wherein at least one of the compounds is a co-crystal former and the other is Compound 5. Weak interactions are defined as interactions that are neither ionic nor covalent, and include, for example: hydrogen bonding, van der Waals forces, and π-π interactions.

[0223] The term "amorphous form" or "amorphous" refers to a material that lacks long range order and therefore does not exhibit distinct X-ray diffraction peaks (ie, Bragg diffraction peaks). An XRPD pattern of an amorphous material is characterized by one or more amorphous halos.

[0224] The term "amorphous halo" is the approximately bell-shaped maximum in the X-ray powder pattern of an amorphous material.

[0225] The term "substantially pure" refers to a solid form of Compound 9 having a purity greater than about 95%, specifically greater than about 99.5%, more specifically greater than about 99.8%, and still more specifically greater than about 99.9%.

[0226] The term "patient" refers to animals, such as dogs, cats, cows, horses, sheep and humans. Particular patients are mammals. The term patient includes males and females.

[0227] The terms "treating," "treat," or "treatment" and such include preventative (eg, prophylactic) and palliative treatment.

[0228] The term "excipient" means any pharmaceutically acceptable additive, carrier, diluent, adjuvant or other ingredient other than the active pharmaceutical ingredient (API) that is typically incorporated for formulation and / or administration to a patient.

[0229] Pharmaceutical compositions, administration and routes of administration

[0230] Also provided herein are pharmaceutical compositions comprising compounds as disclosed herein and pharmaceutically acceptable excipients, such as diluents or carriers. Compounds and pharmaceutical compositions suitable for use in the present invention include those that can be administered in effective amounts to achieve their intended purpose. Administration of the compounds is described in more detail below.

[0231] Suitable pharmaceutical formulations can be determined by technicians based on the route of administration and the desired dosage. See, for example, Remington's Pharmaceutical Sciences, 1435-712 (18th edition, Mack Publishing Co, Easton, Pennsylvania, 1990). Formulations can affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the administered medicament. Depending on the route of administration, suitable dosages can be calculated based on body weight, body surface area, or organ size. Those of ordinary skill in the art, without excessive experimentation, will routinely perform further refinements of the calculations required to determine appropriate therapeutic dosages, particularly based on the dosage information and assays disclosed herein and the pharmacokinetic data available through animal or human clinical trials.

[0232] The phrase "pharmaceutically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse reactions, allergic reactions or other untoward reactions when administered to animals or humans. As used herein, "pharmaceutically acceptable" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. The use of such excipients for pharmaceutically active substances is well known in the art. Unless any conventional media or agents are incompatible with the therapeutic composition, they are considered for use in the therapeutic composition. Supplementary active ingredients may also be incorporated into the composition. In an exemplary embodiment, the formulation may include corn syrup solids, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tricalcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dipotassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, Calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, ketone sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3 and cyanocobalamin.

[0233] The compound may be present in the pharmaceutical composition as a pharmaceutically acceptable salt. As used herein, "pharmaceutically acceptable salt" includes, for example, base addition salts and acid addition salts.

[0234] Pharmaceutically acceptable base addition salts can be formed with metals or amines (e.g., alkali metals and alkaline earth metals or organic amines). Pharmaceutically acceptable salts of compounds can also be prepared with pharmaceutically acceptable cations. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkali metal cations, alkaline earth metal cations, ammonium cations, and quaternary ammonium cations. Carbonates or bicarbonates are also possible. Examples of metals used as cations are sodium, potassium, magnesium, ammonium, calcium, or ferric iron, etc. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine.

[0235] Pharmaceutically acceptable acid addition salts include inorganic or organic acid salts. Examples of suitable acid salts include hydrochlorides, formates, acetates, citrates, salicylates, nitrates, phosphates. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include, for example, formic acid, acetic acid, citric acid, oxalic acid, tartaric acid or mandelic acid, hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic formic acid, sulfonic acid, sulfonic acid or phosphoric acid or N-substituted aminosulfonic acid, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2- Salts of phenoxybenzoic acid, 2-acetoxybenzoic acid, pamoic acid, nicotinic acid or isonicotinic acid; and salts with amino acids, such as the 20 alpha amino acids involved in protein synthesis in nature, such as glutamic acid or aspartic acid, and with phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane 1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene 2-sulfonic acid, naphthalene 1,5-disulfonic acid, 2-phosphoglyceric acid or 3-phosphoglyceric acid, glucose 6-phosphate, N-cyclohexylaminosulfonic acid (for the formation of cyclamates), or with other acidic organic compounds, such as ascorbic acid.

[0236] Pharmaceutical compositions containing the compounds disclosed herein can be manufactured in a conventional manner, eg, by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation depends upon the route of administration chosen.

[0237] For oral administration, suitable compositions can be easily prepared by combining the compounds disclosed herein with pharmaceutically acceptable excipients (e.g., carriers) well known in the art. Such excipients and carriers enable the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by patients to be treated. Pharmaceutical preparations for oral use can be obtained in the following manner: solid excipients are added to the compounds disclosed herein, the resulting mixture is optionally ground, and the granular mixture is processed (if necessary) after adding suitable adjuvants to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. If necessary, disintegrants can be added. Pharmaceutically acceptable ingredients for various types of preparations are well known, and can be, for example, adhesives (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coating materials, preservatives, dyes, thickeners, adjuvants, antimicrobial agents, antioxidants, and carriers for various types of preparations.

[0238] When a therapeutically effective amount of a compound disclosed herein is administered orally, the composition is typically in the form of a solid (eg, tablet, capsule, pill, powder or lozenge) or liquid formulation (eg, aqueous suspension, solution, elixir or syrup).

[0239] When administered in tablet form, the composition may additionally contain functional solids and / or solid carriers, such as gelatin or adjuvants.Tablets, capsules and powders may contain from about 1% to about 95% of the compound, and preferably from about 15% to about 90% of the compound.

[0240] When applied in liquid or suspension form, functional liquids and / or liquid carriers, such as water, petroleum, or oils of animal or plant origin, may be added. The liquid form of the composition may further contain physiological saline solution, sugar alcohol solution, dextrose or other sugar solution or glycol. When applied in liquid or suspension form, the composition may contain about 0.5% to about 90% of the compound disclosed herein by weight, and preferably about 1% to about 50% of the compound disclosed herein. In one embodiment considered, the liquid carrier is non-aqueous or substantially non-aqueous. For application in liquid form, the composition may be supplied as a fast-dissolving solid formulation for dissolving or suspending before being applied.

[0241] When a therapeutically effective amount of the compound disclosed herein is administered by intravenous, percutaneous or subcutaneous injection, the composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution. The preparation of such parenterally acceptable solutions should take into full account pH, isotonicity, stability, etc., and is within the technical scope of the art. In addition to the compounds disclosed herein, preferred compositions for intravenous, percutaneous or subcutaneous injections generally contain isotonic vehicles. Such compositions can be prepared for use as a solution of a free alkali or a pharmacologically acceptable salt in water appropriately mixed with a surfactant (e.g., hydroxypropylcellulose). Dispersions can also be prepared in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as in oils. Under common storage and use conditions, these preparations may optionally contain a preservative to prevent microbial growth.

[0242] Injectable compositions may include sterile aqueous solutions, suspensions or dispersions, and sterile powders for extemporaneous preparation of sterile injectable solutions, suspensions or dispersions. In all embodiments, the form must be sterile, and fluidity must be such that there is easy injectability. It must be stable under manufacturing and storage conditions, and must resist the contamination of microorganisms (e.g., bacteria and fungi) by optionally including preservatives. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. In one embodiment considered, the carrier is non-aqueous or substantially non-aqueous. Appropriate fluidity may be maintained, for example, by using a coating, such as lecithin; by maintaining the desired particle size of the compound in the embodiment of the dispersion; and by using a surfactant. Prevention of microbial action may be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many embodiments, it will be preferred to include an isotonic agent (e.g., sugar or sodium chloride). Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0243] Sterile injectable solutions are prepared by incorporating the active compound in the desired amount into an appropriate solvent optionally containing various other ingredients listed above, followed by sterilization filtration. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile vehicle containing a basic dispersion medium and other desired ingredients from those listed above. In the example of sterile powders for the preparation of sterile injectable solutions, preferred methods of preparation are vacuum drying and freeze drying techniques, which produce a powder of the active ingredient plus any desired other ingredients from a previously sterile filtered solution thereof.

[0244] Slow release or sustained release formulations can also be prepared to achieve controlled release of active compounds in contact with body fluids in the gastrointestinal tract, and provide substantially constant and effective active compound levels in blood plasma. For example, release can be controlled by one or more of dissolution, diffusion and ion exchange. In addition, slow release methods can promote absorption by saturable or restricted access in the gastrointestinal tract. For example, for this purpose, the compound can be embedded in a polymer matrix of a biodegradable polymer, a water-soluble polymer or a mixture of the two and optionally a suitable surfactant. In this case, embedding can mean mixing microparticles in a polymer matrix. Controlled release formulations are also obtained by encapsulating dispersed microparticles or emulsified droplets via known dispersion or emulsion coating techniques.

[0245] For administration by inhalation, the compounds of the invention are conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0246] The compounds disclosed herein can be formulated for parenteral administration by injection (e.g., by bolus or continuous infusion). Injectable formulations can be presented in unit dosage form (e.g., in ampoules or in multi-dose containers) with added preservatives. The composition can take the form of, for example, a suspension, solution, or emulsion in an oily or aqueous vehicle, and can contain preparatants such as suspending agents, stabilizers, and / or dispersants.

[0247] Pharmaceutical formulations for parenteral administration include aqueous solutions of compounds in water-soluble form. In addition, the suspension of the compound can be prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions can contain substances that improve the viscosity of the suspension. Optionally, the suspension can also contain a suitable stabilizer or improve the solubility of the compound and allow the preparation of a highly concentrated solution. Alternatively, the present composition can be in powder form for use before use with a suitable vehicle (e.g., sterile pyrogen-free water) structure.

[0248] The compounds disclosed herein can also be formulated in rectal compositions, such as suppositories or retention enemas (e.g., containing conventional suppository bases). In addition to the formulations described previously, the compounds can also be formulated as long-acting preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Thus, for example, the compounds can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives (e.g., as sparingly soluble salts).

[0249] In particular, the compounds disclosed herein can be administered orally, buccally or sublingually in the form of tablets containing excipients (e.g., starch or lactose), or in capsules or ovules alone or mixed with excipients, or in the form of elixirs or suspensions containing flavoring agents or coloring agents. Such liquid preparations can be prepared with pharmaceutically acceptable additives (e.g., suspending agents). The compounds can also be injected parenterally, e.g., intravenously, intramuscularly, subcutaneously, or intracoronarily. For parenteral administration, the compounds are best used in the form of sterile aqueous solutions, which may contain other substances, such as salts or sugar alcohols (e.g., mannitol) or glucose, to make the solution isotonic with the blood.

[0250] For veterinary use, the compounds disclosed herein are administered as suitably acceptable formulations in accordance with normal veterinary practice. A veterinarian can readily determine the dosage regimen and route of administration most suitable for a particular animal.

[0251] In some embodiments, all required components for treating KRAS-related disorders using compounds as disclosed herein (alone or in combination with another agent or intervention traditionally used to treat such diseases) can be packaged into a kit. Specifically, the present invention provides a kit for therapeutic intervention of a disease, the kit comprising a packaged set of drugs, including compounds disclosed herein and buffers and other components for preparing a deliverable form of the drug; and / or a device for delivering such drugs; and / or any agent for combination therapy with the compounds disclosed herein; and / or instructions for treating the disease packaged with the drug. The instructions can be fixed in any tangible medium, such as printed paper, or computer-readable magnetic or optical media, or instructions referring to a remote computer data source, such as a World Wide Web page accessible via the Internet.

[0252] "Therapeutically effective amount" means an amount that effectively treats or prevents the development of existing symptoms or alleviates existing symptoms in the treated subject. The determination of an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, a "therapeutically effective dose" refers to the amount of a compound that results in the achievement of a desired effect. For example, in a preferred embodiment, a therapeutically effective amount of a compound disclosed herein reduces KRAS activity by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%, compared to a control.

[0253] The amount of compound administered may depend on the subject being treated, the subject's age, health, sex, and weight, the type of concurrent treatment (if any), the severity of the condition, the nature of the desired effect, the manner and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on arterial oxygen tension. However, the most preferred dosage may be adjusted for individual subjects, as will be understood by those skilled in the art and can be determined without undue experimentation. This generally includes adjusting the standard dosage (e.g., if the patient is of low weight, then reducing the dosage).

[0254] Although individual needs vary, determination of the optimal range of effective amounts of the compounds is within the skill of the art. For administration to humans in the curative or prophylactic treatment of the conditions and disorders identified herein, for example, a typical dosage of the compounds of the invention may be from about 0.05 mg / kg / day to about 50 mg / kg / day, for example at least 0.05 mg / kg, at least 0.08 mg / kg, at least 0.1 mg / kg, at least 0.2 mg / kg, at least 0.3 mg / kg, at least 0.4 mg / kg or at least 0.5 mg / kg, and preferably 50 mg / kg or less, 40 mg / kg or less, 30 mg / kg or less, 20 mg / kg or less or 10 mg / kg or less, for example, it may be from about 2.5 mg / day (0.5 mg / kg x 5 kg) to about 5000 mg / day (50 mg / kg x 100 kg). For example, the dosage of the compound can be about 0.1 mg / kg / day to about 50 mg / kg / day, about 0.05 mg / kg / day to about 10 mg / kg / day, about 0.05 mg / kg / day to about 5 mg / kg / day, about 0.05 mg / kg / day to about 3 mg / kg / day, about 0.07 mg / kg / day to about 3 mg / kg / day, about 0.09 mg / kg / day to about 3 mg / kg / day, about 0.05 mg / kg / day to about 0.1 mg / kg / day, about 0.1 mg / kg / day to about 1 mg / kg / day, about 1 mg / kg / day to about 10 mg / kg / day, about 1 mg / kg / day to about 5 mg / kg / day, about 1 mg / kg / day to about 3 mg / kg / day, about 3 mg / day to about 500 mg / day, about 5 mg / day to about 250 mg / day, about 10 mg / day to about 100 mg / day, about 3 mg / day to about 10 mg / day, or about 100 mg / day to about 250 mg / day. Such dosages can be administered in a single dose or can be divided into multiple doses.

[0255] Methods of using KRAS G12C inhibitors

[0256] The present disclosure provides a method for inhibiting RAS-mediated cell signaling, the method comprising contacting a cell with an effective amount of one or more compounds disclosed herein. The inhibition of RAS-mediated signal transduction can be evaluated and confirmed by numerous methods known in the art. Non-limiting examples include showing the following items: (a) a decrease in the GTPase activity of RAS; (b) a decrease in GTP binding affinity or an increase in GDP binding affinity; (c) an increase in the K dissociation of GTP or a decrease in the K dissociation of GDP; (d) a decrease in the level of downstream signaling transduction molecules in the RAS pathway, such as a decrease in the level of pMEK, pERK or pAKT; and / or (e) a decrease in the binding of RAS complexes to downstream signaling molecules (including but not limited to Raf). One or more of the above items can be determined using kits and commercially available assays.

[0257] The disclosure also provides methods of using the compounds or pharmaceutical compositions of the disclosure to treat disease conditions, including but not limited to conditions (e.g., cancer) affected by G12C KRAS, HRAS, or NRAS mutations.

[0258] In some embodiments, a method for treating cancer is provided, comprising administering to a subject in need thereof an effective amount of any of the aforementioned pharmaceutical compositions comprising a compound as disclosed herein. In some embodiments, the cancer is mediated by a KRAS, HRAS or NRAS G12C mutation. In various embodiments, the cancer is pancreatic cancer, colorectal cancer or lung cancer. In some embodiments, the cancer is gallbladder cancer, thyroid cancer and bile duct cancer.

[0259] In some embodiments, the disclosure provides a method of treating a disorder in a subject in need thereof, wherein the method comprises determining whether the subject has a KRAS, HRAS or NRAS G12C mutation, and if the subject is determined to have a KRAS, HRAS or NRAS G12C mutation, administering to the subject a therapeutically effective dose of at least one compound as disclosed herein, or a pharmaceutically acceptable salt thereof.

[0260] The disclosed compounds inhibit anchorage-independent cell growth and thus have the potential to inhibit tumor metastasis. Accordingly, another embodiment of the present disclosure provides a method of inhibiting tumor metastasis, the method comprising administering an effective amount of a compound disclosed herein.

[0261] KRAS, HRAS or NRAS G12C mutations have also been identified in hematological malignancies (e.g., cancers affecting the blood, bone marrow and / or lymph nodes). Therefore, certain embodiments relate to administering the disclosed compounds (e.g., in the form of a pharmaceutical composition) to patients who need to treat hematological malignancies. Such malignancies include, but are not limited to, leukemias and lymphomas. For example, the currently disclosed compounds can be used to treat diseases such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myeloid leukemia (CML), acute monocytic leukemia (AMoL) and / or other leukemias. In other embodiments, these compounds can be used to treat lymphomas, such as all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphoma. In various embodiments, these compounds can be used to treat plasma cell malignancies, such as multiple myeloma, mantle cell lymphoma and Waldenstrom's macroglobulinemia.

[0262] Determining whether a tumor or cancer contains a G12C KRAS, HRAS or NRAS mutation can be performed by evaluating the nucleotide sequence encoding the KRAS, HRAS or NRAS protein, by evaluating the amino acid sequence of the KRAS, HRAS or NRAS protein, or by evaluating the characteristics of a putative KRAS, HRAS or NRAS mutant protein. The sequence of wild-type human KRAS, HRAS or NRAS is known in the art (e.g., Accession No. NP203524).

[0263] Methods for detecting mutations in KRAS, HRAS or NRAS nucleotide sequences are known to those skilled in the art. These methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays, polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) assays, real-time PCR assays, PCR sequencing, mutant allele-specific PCR amplification (MASA) assays, direct sequencing, primer extension reactions, electrophoresis, oligonucleotide ligation assays, hybridization assays, TaqMan assays, SNP genotyping assays, high-resolution melting assays and microarray analysis. In some embodiments, samples are evaluated for G12C KRAS, HRAS or NRAS mutations by real-time PCR. In real-time PCR, a fluorescent probe specific to KRAS, HRAS or NRAS G12C mutations is used. When a mutation is present, the probe binds and detects fluorescence. In some embodiments, a direct sequencing method using a specific region (e.g., exon 2 and / or exon 3) in a KRAS, HRAS or NRAS gene is used to identify KRAS, HRAS or NRAS G12C mutations. This technique will identify all possible mutations in the sequenced region.

[0264] Methods for detecting mutations in KRAS, HRAS or NRAS proteins are known to those skilled in the art. These methods include, but are not limited to, detecting KRAS, HRAS or NRAS mutants using binding agents (e.g., antibodies) specific for the mutant proteins, protein electrophoresis and Western blotting, and direct peptide sequencing.

[0265] Methods for determining whether a tumor or cancer comprises a G12C KRAS, HRAS or NRAS mutation can use a variety of samples. In some embodiments, the sample is taken from a subject with a tumor or cancer. In some embodiments, the sample is a fresh tumor / cancer sample. In some embodiments, the sample is a frozen tumor / cancer sample. In some embodiments, the sample is a formalin-fixed paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

[0266] The present disclosure also relates to a method of treating a hyperproliferative disorder in a mammal, the method comprising administering to the mammal a therapeutically effective amount of a compound as disclosed herein or a pharmaceutically acceptable salt thereof. In some embodiments, the method relates to treating a subject having cancer, such as acute myeloid leukemia, juvenile cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical teratoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem cell glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoid tumor, embryonal tumor, blastoma, primary lymphoma, cervical cancer, childhood cancer, chordoma, heart tumor, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia, CML, chronic myeloproliferative disorder, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumor, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal glioma, Ewing sarcoma, extracranial ectodermal tumor, gonadal ectodermal tumor, eye cancer, bone fibrous histiocytoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), blastoma, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic neuroendocrine Tumors, Kidney Cancer, Laryngeal Cancer, Lip and Oral Cancer, Liver Cancer, Lobular Carcinoma in Situ (LCIS), Lung Cancer, Lymphoma, Metastatic Squamous Neck Cancer with Occult Primary, Midline Cancer, Oral Cancer, Multiple Endocrine Neoplasia Syndrome, Multiple Myeloma / Plasma Cell Neoplasms, Mycosis Fungoides, Myelodysplastic Syndrome, Myelodysplastic / Myeloproliferative Neoplasms, Multiple Myeloma, Merkel Cell Carcinoma, Malignant Mesothelioma, Malignant Fibrous Histiocytoma of Bone and Osteosarcoma, Nasal and Paranasal Sinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Non-Hodgkin Lymphoma, Non-Small Cell Lung Cancer (NSCLC), Mouth Cancer, Lip and Oral Cancer, Oropharyngeal Cancer, Ovarian Cancer, Pancreas In some embodiments, the method is directed to treating a non-cancerous hyperproliferative disorder, such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hypertrophy (BPH)).

[0267] In some embodiments, the methods of treatment relate to treating lung cancer, which methods include administering an effective amount of any of the above compounds (or a pharmaceutical composition comprising the compound) to a subject in need thereof. In certain embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer, or large cell lung cancer. In some embodiments, the lung cancer is small cell lung cancer. Other lung cancers that can be treated with the disclosed compounds include, but are not limited to, adenomas, carcinoid tumors, and undifferentiated carcinomas.

[0268] The present disclosure further provides a method for regulating the activity of G12C mutant KRAS, HRAS or NRAS proteins, which is carried out by contacting the protein with an effective amount of a compound disclosed herein. Regulation can be to inhibit or activate protein activity. In some embodiments, the present disclosure provides a method for inhibiting protein activity, which is carried out by contacting a G12C mutant KRAS, HRAS or NRAS protein with an effective amount of a compound disclosed herein in a solution. In some embodiments, the present disclosure provides a method for inhibiting the activity of G12C mutant KRAS, HRAS or NRAS proteins, which is carried out by contacting cells, tissues or organs expressing a protein of interest. In some embodiments, the present disclosure provides a method for inhibiting protein activity in subjects including but not limited to rodents and mammals (e.g., humans), which is carried out by administering an effective amount of a compound disclosed herein to the subject. In some embodiments, the percentage of regulation exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90%. In some embodiments, the percentage of inhibition exceeds 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.

[0269] In some embodiments, the present disclosure provides a method for inhibiting KRAS, HRAS or NRAS G12C activity in a cell, which is performed by contacting the cell with a certain amount of a compound of the present disclosure, which is sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the cell. In some embodiments, the present disclosure provides a method for inhibiting KRAS, HRAS or NRAS G12C activity in a tissue, which is performed by contacting the tissue with a certain amount of a compound of the present disclosure, which is sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the tissue. In some embodiments, the present disclosure provides a method for inhibiting KRAS, HRAS or NRAS G12C activity in an organism, which is performed by contacting the organism with a certain amount of a compound of the present disclosure, which is sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the organism. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS or NRAS G12C activity in an animal, which is performed by contacting the animal with a certain amount of a compound of the present disclosure, which is sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the animal. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS or NRAS G12C activity in a mammal, which is performed by contacting the mammal with a certain amount of a compound of the present disclosure, which is sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the mammal. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS or NRAS G12C activity in humans, which is performed by contacting the human with a certain amount of a compound of the present disclosure, which is sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the human. The present disclosure provides a method of treating a disease mediated by KRAS, HRAS or NRAS G12C activity in a subject in need of such treatment.

[0270] Combination therapy

[0271] The disclosure also provides methods for combination therapy, wherein agents known to modulate other pathways or other components of the same pathway, or even overlapping groups of target enzymes, are used in combination with the compounds of the disclosure or pharmaceutically acceptable salts thereof. In one aspect, such therapy includes, but is not limited to, the combination of one or more compounds of the disclosure with chemotherapeutic agents, therapeutic antibodies, and radiation therapy to provide a synergistic or additive therapeutic effect.

[0272] Many chemotherapeutic agents are currently known in the art and can be used in combination with the compounds of the present disclosure. In some embodiments, the chemotherapeutic agent is selected from the group consisting of: mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules, such as (imatinib mesylate), (carfilzomib), (bortezomib), Casodex (bicalutamide), (gefitinib), Venclexta TM (Venatolla) and Adriamycin TM (doxorubicin) and various chemotherapeutic agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan TM ); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquinone, meturedopa, and uredopa; ethyleneimines and methylamelamines including hexamethylmelamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphaoramide, and trimethylolomelamine; nitrogen mustards such as chlorambucil, naphthyl mustard, clofosfamide, estramustine, ifosfamide, mechlorethamine oxide, mechlorethamine hydrochloride, and chlorambucil. hydrochloride), melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics, such as aclacinomysin, actinomycin, authramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomicin, carzinophilin, Casodex TM, chromomycin, dactinomycin, daunomycin, detoxibicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogamycin, olivemycin, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptomycin, streptozotocin, tuberculocin, ubenimex, zoloft, daunorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs, such as denopt erin), methotrexate, pteropterin, trimetrexate; purine analogs, such as fludarabine, 6-mercaptopurine, thiopurine, thioguanine; pyrimidine analogs, such as ancitabine (azacitidine), azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, androgens, such as captestosterone, dromostanolone propionate, cyclothiodine, melastosane, testolactone; antiadrenal drugs, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as folinic acid (folinic acid), leucovorin, doxycycline, cyclopentyl ether, doxycycline ... acid; aceglucuronide; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; colcemid; diaziquone; elfomithine; elliptonium acetate; etoglu; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidarol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllic acid podophyllinicacid; 2-ethylhydrazide; procarbazine; PSK; aproximine; sizoran; spirogermanamine; tricosporic acid; triazoquinone; 2,2',2"-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; dibromomannitol; dibromodulcitol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxanes, such as paclitaxel and docetaxel; retinoic acid; esperamicin; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

[0273] Suitable chemotherapeutic cell conditioning agents also include antihormonal agents used to modulate or inhibit the effects of hormones on tumors, such as antiestrogens, including, for example, tamoxifen (NolvadexTM ), raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, troxifene, keoxifene, LY 117018, onapristone, and toremifene (Falenton); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO).

[0274] If desired, the compounds or pharmaceutical compositions disclosed herein may be used in combination with commonly prescribed anticancer drugs, e.g. ABVD, AVICINE, Abagovomab, Acridine carboxamide, Adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, Alpharadin, Alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazide, Amonafide, Anthracenedione, Anti-CD22 immunotoxin, Antineoplastic drugs, Antitumorigenic herbs, Apaziquone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW 2992, Biricodar, Brostallicin, Bryostatin, Buthionine Sulfoxidesulfoximine), CBV (chemotherapy), Calyculin, Cell cycle nonspecific antineoplastic agents, Dichloroacetic acid, Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin, Everolimus, Exatecan, Exisulind, Ferruginol, Forodesine, Fosfestrol, ICE chemotherapy regimen, IT-101, Imexon, Imiquimod, Indolocarbazole, Irov Irofulven, Laniquidar, Larotaxel, Lenalidomide, Lucanthone, Lurtotecan, Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Ortataxel, PAC-1, Pawpaw, Pixantrone, Proteasome Inhibitors, Rebeccamycin, Resiquimod, Rubitecan, SN-38, Salinosporamide A A), Sapacitabine, Stanford V, Swainsonine, Talaporfin, Tariquidar, Tegafur-uracil, Temodar, Tesetaxel, Triplatin tetranitrate, Tris(2-chloroethyl)amine, Troxacitabine, Uramustine, Vadimezan, Vinflunine, ZD6126, or Zosuquidar.

[0275] The disclosure further relates to methods of inhibiting abnormal cell growth or treating hyperproliferative disorders in mammals using a combination of a compound or pharmaceutical composition provided herein and radiotherapy. Techniques for administering radiotherapy are known in the art, and these techniques can be used in the combination therapies described herein. The administration of the disclosed compound in such a combination therapy can be determined as described herein.

[0276] Radiotherapy can be applied by one of several methods or a combination of methods, including but not limited to external beam therapy, internal radiotherapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy and permanent or temporary interstitial brachytherapy. As used herein, the term "brachytherapy" refers to radiotherapy delivered by a spatially confined radioactive material that is inserted into the body at or near a tumor or other proliferative tissue diseased site. The term is intended to include exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32 and Lu radioisotopes) without limitation. Suitable radioactive sources used as cell conditioning agents of the present disclosure include solids and liquids. By way of non-limiting example, the radiation source can be a radionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation or other therapeutic rays. The radioactive material can also be a fluid prepared from any solution of one or more radionuclides (e.g., a solution of I-125 or I-131), or the radioactive fluid can be produced using a slurry of a suitable fluid containing small particles of solid radionuclides (e.g., Au-198, Y-90). In addition, one or more radionuclides can be embedded in a gel or radioactive microspheres.

[0277] The compounds or pharmaceutical compositions disclosed herein may be used in combination with a certain amount of one or more substances selected from the following: anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolysis inhibitors, or autophagy inhibitors.

[0278] Anti-angiogenic agents can be used in conjunction with the compounds disclosed herein and the pharmaceutical compositions described herein, and these anti-angiogenic agents are, for example, MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, and COX-11 (cyclooxygenase 11) inhibitors. Anti-angiogenic agents include, for example, rapamycin, temsirolimus (temsirolimus, CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in the following patents: WO 96 / 33172, WO 96 / 27583, European Patent Publication EP 0818442, European Patent Publication EP1004578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, WO 98 / 34915, WO 98 / 33768, WO 98 / 30566, European Patent Publication 606046, European Patent Publication 931 788, WO 90 / 05719, WO 99 / 52910, WO 99 / 52889, WO 99 / 29667, WO 1999007675, European Patent Publication EP 1786785, European Patent Publication No. EP 1181017, U.S. Patent Publication No. US 20090012085, US Publication US 5863 949, US Publication US 5861510 and European Patent Publication EP 0780386, all of which are incorporated herein by reference in their entirety. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity in inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and / or AMP-9 relative to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12 and MMP-13). Some specific examples of MMP inhibitors that can be used in the present disclosure are AG-3340, RO 32-3555 and RS 13-0830.

[0279] The compounds of the invention may also be used in co-therapy with other antineoplastic agents, such as acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, hexamethylmelamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM 002 (Novelos), bexarotene, bicalutamide, bromuridine, capecitabine, cimoleukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030 (Dong-A), daclizumab, denileukin diftitox), deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon-α, daunorubicin, doxorubicin, retinoic acid, edilofosine, edrecolomab, eflornithine, ethidium bromide, epirubicin, erythropoietin beta, etoposide phosphate, exemestane, exisulind, fadroxil, filgrastim, non nasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin, gimeracil / oteracil / tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronate, idarubicin, imiquimod, interferon-α, interferon-α, natural interferon-α-2, interferon-α-2a, interferon-α-2b, interferon-α-N1, interferon-α- n3, interferon alfacon-1, interferon alpha, natural interferon beta, interferon beta-1a, interferon beta-1b, interferon gamma, natural interferon gamma-1a, interferon gamma-1b, interleukin-1β, iodobenzylguanidine, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, interferon alpha, leuprolide, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprofen, melarsoprol, metoclopramide, mifepristone, miltefosine, milistostim, mismatched double-stranded RNA, mitoguanidine, dibromodulanol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, narcotine, novel erythropoiesis stimulating protein, NSC 631570Octreotide, oprelvekin, osephatron, oxaliplatin, paclitaxel, pamidronate, pegaspargase, peginterferon-α-2b, pentosan polysulfatesodium, pentostatin, picibanil, pirarubicin, rabbit anti-thymocyte polyclonal antibody, peginterferon-α-2a, porfimer sodium, raloxifene, raltitrexed, rasburiembodiment, etidronate rhenium (Re 186), RII retinoic acid phenamide (RIIretinamide), rituximab, romotide, samarium (153Sm) lexidronam, sargramostim, sizoran, sobuzosine, sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymofasin, thyrotropin alfa, topotecan can), toremifene, tositumomab-iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor alpha, natural ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulizin, zinostatinstimalamer, or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotides, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diazocone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocibine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEAMAb (Trilex), LYM-1-iodine 131 MAb (Techniclone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menolide, mitumomab, motexafingadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), lupinotecan, satraplatin, sodium phenylacetate, sparlfosicacid, SRL 172 (SR Pharma), SU 5416 (SUGEN, now Pfizer, Inc.), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyletiopurpurin, tirapazamine, cancer vaccine (Bamira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), melanoma tumor lysate vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or valspodar.

[0280] The compounds of the present invention may further be used together with a VEGFR inhibitor. Other compounds described in the following patents and patent applications may be used in combination therapy: US 6,258,812, US 2003 / 0105091, WO 01 / 37820, US 6,235,764, WO 01 / 32651, US 6,630,500, US 6,515,004, US 6,713,485, US 5,521,184, US 5,770,599, US 5,747,498, WO 02 / 68406, WO 02 / 66470, WO 02 / 55501, WO 04 / 05279, WO 04 / 07481, WO 04 / 07458, WO 04 / 09784, WO 02 / 59110, WO 99 / 45009, WO 00 / 59509, WO 99 / 61422, US 5,990,141, WO 00 / 12089 and WO 00 / 02871.

[0281] In some embodiments, the combination comprises a combination of the composition of the invention and at least one anti-angiogenic agent. Agents include, but are not limited to, chemical compositions, antibodies, antigen binding regions, radionuclides, and combinations and conjugates thereof prepared synthetically in vitro. Agents may be agonists, antagonists, allosteric modulators, toxins, or more generally, may be used to inhibit or stimulate their targets (e.g., receptor or enzyme activation or inhibition), and thereby promote cell death or prevent cell growth.

[0282] Exemplary anti-angiogenic agents include ERBITUX TM (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., antibodies or antigen binding regions that specifically bind to VEGF, or soluble VEGF receptors or their ligand binding regions) (e.g., AVASTIN TM VEGF-TRAP TM ) and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind thereto), EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind thereto) (e.g., Vectibix) panitumumab), IRESSA TM (gefitinib), TARCEVA TM(erlotinib), anti-Ang1 agents and anti-Ang2 agents (e.g., antibodies or antigen binding regions that specifically bind to them or to their receptors (e.g., Tie2 / Tek)) and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind to them). The pharmaceutical compositions of the present invention may also include one or more agents (e.g., antibodies, antigen binding regions or soluble receptors) that specifically bind to growth factors and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor), and antibodies or antigen binding regions that specifically bind to its receptor "c-met".

[0283] Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., U.S. Publication No. 2003 / 0162712; U.S. Pat. No. 6,413,932), anti-TWEAK agents (e.g., specific binding antibodies or antigen binding regions, or soluble TWEAK receptor antagonists; see Wiley, U.S. Pat. No. 6,727,225), ADAM disintegrin domains for antagonizing the binding of integrins to their ligands (Fanslow et al., U.S. Publication No. 2002 / 00 42368), specifically binding anti-eph receptor and / or anti-ephrin antibodies or antigen binding regions (U.S. Patent Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124 and members of their patent families), and anti-PDGF-BB antagonists (e.g., specifically binding antibodies or antigen binding regions), as well as antibodies or antigen binding regions that specifically bind to PDGF-BB ligands and PDGFR kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind thereto).

[0284] Other anti-angiogenic / anti-tumor agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptaniboctasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, US 5712291); Ilomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US5792783); vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA, now known as CASI Pharmaceuticals); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); α-D148Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands); DAC: anti-angiogenic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko Kogyo Co., Ltd., Japan) Hakko); SU-0879 (Pfizer, USA); CGP-79787 (Novartis Pharmaceuticals, Switzerland, EP 970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen-E fragment (Bacta, UK); Angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (Antrimed, USA, now known as CASI Pharmaceuticals); Angiogenesis inhibitor (Tripep, Sweden); Maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences, USA) Corporation);ER-68203-00 (IVAX, USA); fluazifop (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN) (Genentech, USA); angiogenesis inhibitor (Sugen, USA); XL 784 (Exelixis, USA); XL 647 (Excelix, USA); second-generation MAb α5β3 integrin (Applied Molecular Evolution, USA and MedImmune, USA); gene therapy for retinopathy (Oxford BioMedica, UK); enzastaurin hydrochloride (USAN) (Lilly, USA); CEP 7055 (Cerfaron, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); angiogenesis inhibitors (Alchemia, Australia); VEGF antagonists (Regeneron, USA); rBPI 21 and BPI-derived anti-angiogenic agents (XOMA, USA); PI 88 (Progen, Australia); cilengitide (pINN) (Merck, Germany; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); cetuximab (INN) (Aventis, France); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292 (Telios, USA); endostatin (Boston Children's Hospital, USA);ATN 161 (Attenuon, USA); angiostatin (Children's Hospital, Boston, USA); 2-methoxyestradiol (Children's Hospital, Boston, USA); ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); vatalanib (pINN) (Novartis Pharmaceuticals, Switzerland and Schering AG, Germany) AG); tissue factor pathway inhibitor (Antrimed, Inc., USA); pegaptanib (Pinn) (Gilead Sciences, USA); xanthorrhizol (Yonsei University, South Korea); gene-based VEGF-2 vaccine (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX 103 (University of California, San Diego, USA); PX 478 (ProlX, USA); metastasis inhibitor (Antrimed, USA); troponin I (Harvard University, USA); SU 6668 (SUGEN, USA, now known as Pfizer, Inc.); OXI 4503 (OXiGENE, USA); orthoguanidine (Dimensional Pharmaceuticals, USA) =Pharmaceuticals); motuporamine C (British Columbia University, Canada); CDP 791 (Celltech Group, UK); atimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Eterna, Canada); angiogenesis vaccine (Antrimed, USA, now known as CASI Pharmaceuticals); urokinase plasminogen activator inhibitor (Dandelion, USA); oglufanide (pINN) (Melmotte, USA); HIF-1α inhibitor (Xenova, UK); CEP 5214 (Cerfalon, USA); BAY RES 2622 (Bayer, Germany);Angiside (Incoin, USA); A6 (Angstrom, USA); KR 31372 (Korea Research Institute of Chemical Technology, South Korea); GW 2286 (GlaxoSmithKline, UK); EHT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN633 (Kirin Brewery, Japan); intraocular 2-methoxyestradiol drug delivery system (Antrimed, USA); anginex (Maastricht University, Netherlands and Minnesota University, USA); ABT 510 (Abbott Laboratories, USA); AAL 993 (Novartis Pharmaceuticals, Switzerland); VEGI (ProteomTech, USA); TNF-α inhibitor (National Institute on Aging, USA); SU 11248 (Pfizer, USA and Sugen, USA); ABT 518 (Abbott Laboratories, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital and Antrimed, USA); MAb, KDR (ImClone Systems, USA); MAb α5β1 (Protein Design, USA); KDR kinase inhibitor (Cell Technology Group, UK and Johnson & Johnson, USA); GFB 116 (South Florida University and Yale University, USA); CS 706 (Sankyo, Japan); combretastatin A4 prodrug (Arizona State University, USA); chondroitinase AC (IBEX, Canada); BAYRES 2690 (Bayer, Germany); AGM1470 (Harvard University, USA, Takeda Company, Japan, and TAP, USA); AG 13925 (Agouron, USA); tetrathiomolybdate (University of Michigan, USA);GCS 100 (Wayne State University, USA); CV 247 (Ivy Medical, UK); CKD 732 (ChongKun Dang, South Korea); vascular endothelial growth factor MAb (Synova, UK); irsogladine (INN) (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); squalamine (pINN) (Genaera, USA); RPI 4610 (Sirna, USA); cancer therapy (Marinova, Australia); heparanase inhibitor (InSight, Israel); KL 3106 (Kolon, South Korea); honokiol (Emory University, USA). =University); ZK CDK (Schering AG, Germany); ZKAngio (Schering AG, Germany); ZK 229561 (Novartis Pharmaceuticals AG, Switzerland and Schering AG, Germany); XMP 300 (Xiao Ma, USA); VGA 1102 (Taisho, Japan); VEGF receptor modulators (Pharmacopeia, United States); VE-cadherin-2 antagonists (Imclone Systems, USA); Vasostatin (National Institutes of Health, USA); Flk-1 vaccine (Imclone Systems, USA); TZ 93 (Tsumura Co., Ltd., Japan); Tumorstatin (Beth Israel Hospital, USA); Truncated soluble FLT 1 (vascular endothelial growth factor receptor 1) (Merck & Co., USA); Tie-2 ligand (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA). ;

[0285] Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil TM), bafilomycin A1, 5-amino-4-imidazolecarboxamide ribonucleoside (AICAR), okadaic acid, autophagy-inhibiting algal toxins that inhibit type 2A or type 1 protein phosphatase, cAMP analogs, and drugs that increase cAMP levels such as adenosine, LY204002, N6-mercaptopurine ribonucleoside, and vinblastine. In addition, antisense or siRNA that inhibits protein expression can also be used, and these proteins include but are not limited to ATG5 (which is involved in autophagy).

[0286] Other pharmaceutically active compounds / agents that can be used to treat cancer and can be used in combination with one or more compounds of the present invention include: erythropoietin alfa; darbepoetin alfa; panitumumab; pegfilgrastim; palifermin; filgrastim; denosumab; anciplostim; AMG 102; AMG 176; AMG 386; AMG 479; AMG 655; AMG 745; AMG 951; and AMG 706, or pharmaceutically acceptable salts thereof.

[0287] In certain embodiments, the compositions provided herein are administered in combination with a chemotherapeutic agent. Suitable chemotherapeutic agents may include natural products, such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), taxol, epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunomycin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparaginase, which systemically metabolizes L-asparagine and deprive cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative / antimitotic alkylating agents (e.g., nitrogen mustards, e.g., dichloromethyl diethylamide, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimines and methylmelamines (e.g., hexamethylmelaamine and thiotepa), CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-03 32991, dinaciclib, P27-00, AT-7519, RGB286638 and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs and streptozotocin), trazenes-dacarbazinine (DTIC), antiproliferative / antimitotic antimetabolites such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., The drugs include 5-fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoylanilide hydroxamic acid, acid, vorinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP (Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g., Zalypsis), PI3Kδ inhibitors (e.g., GS-1101 and TGR-1202), PI3Kδ and γ inhibitors (e.g., CAL-130), multikinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g.,estrogen) and hormone agonists such as luteinizing hormone releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38 MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN 163L), Aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CS1 (e.g., elotuzumab), HSP90 inhibitors (e.g., 17AAG and KOS 953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTIs (e.g., Zarnestra, TM ), anti-CD138 (e.g., BT062), Torc1 / 2 specific kinase inhibitors (e.g., INK128), kinase inhibitors (e.g., GS-1101), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), BCL-2 antagonists. Other chemotherapeutic agents may include dichloromethyl diethylamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib or any analog or derivative variant of the foregoing.

[0288] The compounds of the present invention may also be used in combination with radiation therapy, hormone therapy, surgery and immunotherapy, which are well known to those skilled in the art.

[0289] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with steroids. Suitable steroids may include, but are not limited to, 21-acetoxypregnenolone, alclomethasone, algestrone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazole, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difuprednate, glycyrrhetinic acid, fluzacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinolone acetate, fluocortinbutyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, hydrocortisone, loteprednol etabonate, maprednione, medrysone, methylprednisolone, methylprednisolone, mometasone furoate furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylamino acetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide and their salts and / or derivatives. In a specific embodiment, the compounds of the present invention can also be used in combination with other pharmaceutically active agents for treating nausea.Examples of agents that may be used to treat nausea include: dronabinol; granisetron; metoclopramide; ondansetron; and prochlorperazine; or pharmaceutically acceptable salts thereof.

[0290] The compounds of the present invention can also be used in combination with other pharmaceutically active compounds that destroy or inhibit RAS-RAF-ERK or PI3K-AKT-TOR signaling pathways. In other such combinations, other pharmaceutically active compounds are PD-1 and PD-L1 antagonists. The compounds or pharmaceutical compositions disclosed herein can also be used in combination with a certain amount of one or more selected from the following substances: EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, Mcl-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors and immunotherapy, including monoclonal antibodies, immunomodulatory imides (IMiD), anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1 and anti-OX40 agents, GITR agonists, CAR-T cells and BiTE.

[0291] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNA. Useful EGFR antibody inhibitors include cetuximab (Erbitux), panitumumab (Victobi), zalutumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and the most recent lapatinib (TykerB). See, e.g., Yan L et al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005; 39(4): 565-8, and Paez JG et al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004; 304(5676): 1497-500.

[0292] Non-limiting examples of small molecule EGFR inhibitors include any EGFR inhibitor described in the following patent publications, and all pharmaceutically acceptable salts and solvates of the EGFR inhibitors: European Patent Application EP520722, published on December 30, 1992; European Patent Application EP 566226, published on October 20, 1993; PCT International Publication WO 96 / 33980, published on October 31, 1996; U.S. Patent No. 5,747,498, issued on May 5, 1998; PCT International Publication WO 96 / 30347, published on October 3, 1996; European Patent Application EP 787772, published on August 6, 1997; PCT International Publication WO 97 / 30034, published on August 21, 1997; PCT International Publication WO 97 / 30044, published on August 21, 1997; PCT International Publication WO 97 / 38994, published on October 23, 1997; PCT International Publication WO 97 / 49688, published on December 31, 1997; European Patent Application EP 837063, published on April 22, 1998; PCT International Publication WO 98 / 02434, published on January 22, 1998; PCT International Publication WO 97 / 38983, published on October 23, 1997; PCT International Publication WO 95 / 19774, published on July 27, 1995; PCT International Publication WO 95 / 19970, published on July 27, 1995; PCT International Publication WO 97 / 13771, published on April 17, 1997; PCT International Publication WO 98 / 02437, published on January 22, 1998; PCT International Publication WO 98 / 02438, published on January 22, 1998; PCT International Publication WO 97 / 32881, published on September 12, 1997; German Application DE 19629652, published on January 29, 1998; PCT International Publication WO 98 / 33798, published on August 6, 1998; PCT International Publication WO 97 / 32880, published on September 12, 1997; PCT International Publication WO 97 / 32880, published on September 12, 1997; European Patent Application EP 682027, published on November 15, 1995; PCT International Publication WO 97 / 02266, published on January 23, 197; PCT International Publication WO 97 / 27199, published on July 31, 1997; PCT International Publication WO 98 / 07726, published on February 26, 1998; PCT International Publication WO 97 / 34895, published on September 25, 1997;PCT International Publication WO 96 / 31510', published on October 10, 1996; PCT International Publication WO 98 / 14449, published on April 9, 1998; PCT International Publication WO 98 / 14450, published on April 9, 1998; PCT International Publication WO 98 / 14451, published on April 9, 1998; PCT International Publication WO 95 / 09847, published on April 13, 1995; PCT International Publication WO 97 / 19065, published on May 29, 1997; PCT International Publication WO 98 / 17662, published on April 30, 1998; U.S. Patent No. 5,789,427, issued on August 4, 1998; U.S. Patent No. 5,650,415, issued on July 22, 1997; U.S. Patent No. 5,656,643, issued on August 12, 1997; PCT International Publication WO 99 / 35146, published on July 15, 1999; PCT International Publication WO 99 / 35132, published on July 15, 1999; PCT International Publication WO 99 / 07701, published on February 18, 1999; and PCT International Publication WO 92 / 20642, published on November 26, 1992. Other non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12): 1599-1625. ;

[0293] Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block activation of EGFR by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in the following literature: Modjtahedi, H. et al., 1993, Br. J. Cancer [British Journal of Cancer] 67: 247-253; Teramoto, T. et al., 1996, Cancer [Cancer] 77: 639-645; Goldstein et al., 1995, Clin. Cancer Res. [Clinical Cancer Research] 1: 1311-1318; Huang, SM et al., 1999, Cancer Res. [Cancer Research] 15: 59 (8): 1935-40; and Yang, X. et al., 1999, Cancer Res. [Cancer Research] 59: 1236-1243. Therefore, the EGFR inhibitor may be the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra), or Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment having the binding specificity thereof.

[0294] KRAS of the present invention G12C Inhibitors may be used in combination with MEK inhibitors. Specific MEK inhibitors that may be used in the combinations of the invention include PD-325901, trametinib, pimacetinib, MEK162 [also known as binitinib], TAK-733, GDC-0973, and AZD8330. G12C The specific MEK inhibitor used with the inhibitor is trametinib (trade name: , available from Novartis Pharmaceuticals Corp.). Another specific MEK inhibitor is N-(((2R)-2,3-dihydroxypropyl)oxy)-3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)benzamide, also known as AMG 1009089, 1009089 or PD-325901. Another specific MEK inhibitor that can be used in the combination of the present invention includes cobimetinib. MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886 and ARRY-438162.

[0295] PI3K inhibitors include, but are not limited to, wortmannin, 17-hydroxywortmannin analogs described in WO 06 / 044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as GDC 0941 and described in PCT Publication Nos. WO 09 / 036,082 and WO 09 / 055,730), 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT Publication No. WO 09 / 055,730). 06 / 122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in PCT Publication No. WO 2008 / 070740), LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, available from Axon Medchem), PI103 hydrochloride (3-[4-(4-morpholinopyrido-[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]phenol hydrochloride, available from Axon Medchem), PIK 75 (N'-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfonyl-hydrazide hydrochloride, available from Exon Medical Chemicals), PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)-nicotinamide, available from Exon Medical Chemicals), GDC-0941 dimesylate (2-(1H-indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine dimesylate, available from Exon Medical Chemicals), AS-2524 24 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione, available from Exon Medical Chemicals) and TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one, available from Exon Medical Chemicals), XL-765 and XL-147.Other PI3K inhibitors include demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0296] AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and 2) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05011700); indole-3-carbinol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J. Nutr. [Nutr.] 134(12 Suppl.), 3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; Dasmahapatra et al. (2004) Clin. Cancer Res. [Clinical Cancer Research] 10(15), 5242-52, 2004); phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs [Expert Review of Research Drugs] 13, 787-97); and triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al. (2004) Cancer Res. [Cancer Research] 64, 4394-9).

[0297] TOR inhibitors include, but are not limited to, AP-23573, CCI-779, everolimus, RAD-001, rapamycin, temsirolimus, ATP-competitive TORC1 / TORC2 inhibitors including PI-103, PP242, PP30, and Torin 1. Other TOR inhibitors include FKBP12 enhancers; rapamycin and its derivatives, including: CCI-779 (temsirolimus), RAD001 (everolimus; WO 9409010) and AP23573; rapamycin analogs (rapalogs), such as disclosed in WO 98 / 02441 and WO01 / 14387, such as AP23573, AP23464 or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropionate]-rapamycin (also known as CC1779), 40-epi-(tetrazolyl)-rapamycin (also known as ABT578), 32-deoxyrapamycin, 16-pentynyloxy-32(S)-dihydrorapamycin and WO 98 / 02441 and WO 01 / 14387. 05005434; derivatives disclosed in the following patents: U.S. Patent No. 5,258,389, WO 94 / 090101, WO 92 / 05179, U.S. Patent No. 5,118,677, U.S. Patent No. 5,118,678, U.S. Patent No. 5,100,883, U.S. Patent No. 5,151,413, U.S. Patent No. 5,120,842, WO 93 / 111130, WO 94 / 02136, WO 94 / 02485, WO 95 / 14023, WO94 / 02136, WO 95 / 16691, WO 96 / 41807, WO 96 / 41807 and U.S. Patent No. 5,256,790; phosphorus-containing rapamycin derivatives (e.g., WO 05016252); 4H-1-benzopyran-4-one derivatives (eg, U.S. Provisional Application No. 60 / 528,340).

[0298] MCl-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. Myeloid cell leukemia-1 (MCL-1) protein is one of the key anti-apoptotic members of the B-cell lymphoma 2 (BCL-2) protein family. Overexpression of MCL-1 is closely associated with tumor progression and resistance not only to traditional chemotherapy but also to targeted therapies including BCL-2 inhibitors (e.g., ABT-263).

[0299] In the present invention, KRAS G12CInhibitors may also be used in combination with SHP2 inhibitors. SHP2 inhibitors that may be used in the combinations of the invention include, but are not limited to, SHP099, and RMC-4550 or RMC-4630 (from Revolutions Medicines, Redwood City, CA).

[0300] Proteasome inhibitors include but are not limited to (Carfilzomib), (bortezomib) and oprozomib.

[0301] Immunotherapy includes, but is not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents.

[0302] Monoclonal antibodies include but are not limited to (daratumumab), (trastuzumab), (bevacizumab), (rituximab), (ranibizumab), and (aflibercept).

[0303] Immunomodulatory drugs (IMiDs) are a class of immunomodulatory drugs (drugs that modulate immune responses) that contain an imide group. The IMiD class includes thalidomide and its analogs (lenalidomide, pomalidomide, and amitrasat).

[0304] Anti-PD-1 inhibitors including but not limited to antibodies including but not limited to pembrolizumab and nivolumab Exemplary anti-PD-1 antibodies and methods of using them are described in Goldberg et al., Blood 110(1):186-192 (2007); Thompson et al., Clin. Cancer Res. 13(6):1757-1761 (2007); and Korman et al., International Application No. PCT / JP2006 / 309606 (Publication No. WO2006 / 121168A1), each of which is expressly incorporated herein by reference. TM(ipilimumab) or tremelimumab (targeting CTLA-4), galiximab (targeting B7.1), BMS-936558 (targeting PD-1), MK-3475 (targeting PD-1), AMP224 (targeting B7DC), BMS-936559 (targeting B7-H1), MPDL3280A (targeting B7-H1), MEDI-570 (targeting ICOS), AMG557 (targeting B7H2), MGA271 (targeting B7H3), IMP321 (targeting LAG-3), BMS-663513 (targeting CD137), PF-05082566 (targeting CD137), CDX-1127 (targeting CD27), anti-OX40 (Providence Health Services Services), huMAbOX40L (for OX40L), Atacicept (for TACI), CP-870893 (for CD40), Lucatumumab (for CD40), Dacilizumab (for CD40), Muromonab-CD3 (for CD3), Ipilimumab (for CTLA-4). Immunotherapy also includes genetically engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTE).

[0305] GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as the GITR fusion proteins described in U.S. Pat. No. 6,111,090 box.c, European Patent No.: 090505B1, U.S. Pat. No. 8,586,023, PCT Publication Nos.: WO 2010 / 003118 and 2011 / 090754, or anti-GITR antibodies such as described in U.S. Pat. No. 7,025,962, European Patent No.: 1947183B1, U.S. Pat. No. 7,812,135, U.S. Pat. No. 8,388,967, U.S. Pat. No. 8,591,886, European Patent No.: EP 1866339, PCT Publication No.: WO 2011 / 028683, PCT Publication No.: WO 2013 / 039954, PCT Publication No.: WO 2005 / 007190, PCT Publication No.: WO 2007 / 133822, PCT Publication No.: WO 2005 / 055808, PCT Publication No.: WO 99 / 40196, PCT Publication No.: WO 2001 / 03720, PCT Publication No.: WO 99 / 20758, PCT Publication No.: WO 2006 / 083289, PCT Publication No.: WO 2005 / 115451, U.S. Patent No. 7,618,632 and PCT Publication No.: WO 2011 / 051726.

[0306] The compounds described herein can be used in combination with the medicaments disclosed herein or other suitable medicaments, depending on the condition being treated. Therefore, in some embodiments, one or more compounds of the present disclosure will be co-administered with other medicaments as described above. When used in combination therapy, the compounds described herein are administered simultaneously or separately with the second medicament. This combination administration can include the simultaneous administration of two medicaments in the same dosage form, the simultaneous administration in a separate dosage form, and the separate administration. That is, the compounds described herein and any of the above-mentioned medicaments can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds disclosed herein and any of the above-mentioned medicaments can be administered simultaneously, wherein the two medicaments are present in a separate formulation. In another alternative, any of the above-mentioned medicaments can be administered immediately after the compounds disclosed herein are administered, or vice versa. In some embodiments of the separate administration scheme, the administration of the compounds disclosed herein and any of the above-mentioned medicaments is separated by a few minutes, or a few hours, or a few days.

[0307] Since one aspect of the present invention contemplates treating a disease / disorder with a combination of pharmaceutically active compounds that can be administered separately, the present invention further relates to combining separate pharmaceutical compositions in kit form. The kit comprises two separate pharmaceutical compositions: a compound of the present invention and a second pharmaceutical compound. The kit comprises a container for containing the separate compositions, such as a separate bottle or a separate foil bag. Other examples of containers include syringes, boxes, and bags. In some embodiments, the kit comprises instructions for use of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), when administered at different dosage intervals, or when the prescribing medical professional requires titration of the individual components in the combination.

[0308] All patents and other publications cited herein are incorporated by reference.

[0309] The processes presented below illustrate specific embodiments of the present invention. These processes are representative and are not intended to limit the scope of the claims in any way.

[0310] Related processes of the present invention

[0311] The following intermediate compounds of 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-acryloyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one are representative examples of the present invention and should not be construed as limiting the scope of the present invention.

[0312] The synthesis of compound 9 and related intermediates is described in U.S. Serial No. 15 / 984,855, filed on May 21, 2018, which claims priority to and claims the benefit of U.S. Provisional Application No. 62 / 509,629, filed on May 22, 2017, which are incorporated herein by reference in their entirety for all purposes.

[0313] 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-acryloyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one was prepared using the following method in which the isomers of the final product were separated by chiral chromatography.

[0314]

[0315] Step 1: 2,6-dichloro-5-fluoronicotinamide (Intermediate S). To a mixture of 2,6-dichloro-5-fluoro-nicotinic acid (4.0 g, 19.1 mmol, AstaTech Inc., Bristol, Pennsylvania) in dichloromethane (48 mL) was added oxalyl chloride (2M DCM solution, 11.9 mL, 23.8 mmol), followed by a catalytic amount of DMF (0.05 mL). The reaction was stirred at room temperature overnight and then concentrated. The residue was dissolved in 1,4-dioxane (48 mL) and cooled to 0 °C. Ammonium hydroxide solution (28.0%-30% based on NH3, 3.6 mL, 28.6 mmol) was slowly added via syringe. The resulting mixture was stirred at 0 °C for 30 min and then concentrated. The residue was diluted with a 1:1 mixture of EtOAc / heptane and stirred for 5 min and then filtered. The filtered solid was discarded and the remaining mother liquor was partially concentrated to half volume and filtered. The filtered solid was washed with heptane and dried in a reduced pressure oven (45 °C) overnight to provide 2,6-dichloro-5-fluoronicotinamide. 1 H NMR (400MHz, DMSO-d6) δppm 8.23 ​​(d, J=7.9Hz, 1H) 8.09 (br s, 1H) 7.93 (br s, 1H). m / z (ESI, +ve ion): 210.9 (M+H) + .

[0316] Step 2: 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. Oxalyl chloride (2M DCM solution, 14.4 mL, 28.8 mmol) was slowly added to an ice-cooled slurry of 2,6-dichloro-5-fluoronicotinamide (Intermediate S, 5.0 g, 23.9 mmol) in THF (20 mL) by syringe. The resulting mixture was heated at 75 ° C for 1 h, then heating was stopped and the reaction was concentrated to half volume. After cooling to 0 ° C, THF (20 mL) was added dropwise via cannula, followed by a THF (10 mL) solution of 2-isopropyl-4-methylpyridin-3-amine (Intermediate R, 3.59 g, 23.92 mmol). The resulting mixture was stirred at 0 ° C for 1 h and then quenched with a 1:1 mixture of brine and saturated aqueous ammonium chloride solution. The mixture was extracted with EtOAc (3x), and the combined organic layers were dried over anhydrous sodium sulfate and concentrated to provide 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. This material was used in the following step without further purification. m / z (ESI, +ve ion): 385.1 (M+H) + .

[0317] Step 3: 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione. KHMDS (1M THF solution, 50.2 mL, 50.2 mmol) was slowly added to an ice-cooled solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (9.2 g, 24.0 mmol) in THF (40 mL) via syringe. The ice bath was removed and the resulting mixture was stirred at room temperature for 40 min. The reaction was quenched with saturated aqueous ammonium chloride solution and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH / heptane) to provide 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyridinyl[2,3-d]pyrimidine-2,4(1H,3H)-dione. 1 H NMR(400MHz,DMSO-d6)δppm 12.27(brs,1H),8.48-8.55(m,2H),7.29(d,J=4.8Hz,1H),2.87(quin,J=6.6Hz,1H),1.99-2.06(m,3H),1.09(d,J=6.6Hz,3H),1.01(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d6) δ: -126.90 (s, 1F). m / z (ESI, +ve ion): 349.1 (M+H) + .

[0318] Step 4: 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. To a solution of 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2,4(1H,3H)-dione (4.7 g, 13.5 mmol) and DIPEA (3.5 mL, 20.2 mmol) in acetonitrile (20 mL) was added phosphorus oxychloride (1.63 mL, 17.5 mmol) dropwise via syringe. The resulting mixture was heated at 80 °C for 1 h, then cooled to room temperature and concentrated to provide 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. This material was used in the following step without further purification. m / z (ESI, +ve ion): 367.1 (M+H) + .

[0319] Step 5: (S)-tert-butyl 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. To an ice-cooled solution of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (13.5 mmol) in acetonitrile (20 mL) was added DIPEA (7.1 mL, 40.3 mmol) followed by (S)-4-N-Boc-2-methylpiperazine (3.23 g, 16.1 mmol, Combi-Blocks, Inc., San Diego, CA, USA). The resulting mixture was warmed to room temperature and stirred for 1 h, then diluted with cold saturated aqueous sodium bicarbonate solution (200 mL) and EtOAc (300 mL). The mixture was stirred for an additional 5 min, the layers were separated, and the aqueous layer was extracted with more EtOAc (1x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0%-50% EtOAc / heptane) to provide 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylic acid (S)-tert-butyl ester. m / z (ESI, +ve ion): 531.2 (M+H) + .

[0320] Step 6: (3S)-tert-butyl 4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. A mixture of (S)-tert-butyl 4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (4.3 g, 8.1 mmol), potassium trifluoro(2-fluoro-6-hydroxyphenyl)borate (Intermediate Q, 2.9 g, 10.5 mmol), potassium acetate (3.2 g, 32.4 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) complexed with dichloromethane (661 mg, 0.81 mmol) in 1,4-dioxane (80 mL) was degassed with nitrogen for 1 min. Deoxygenated water (14 mL) was added and the resulting mixture was heated at 90° C. for 1 h. The reaction was cooled to room temperature, quenched with half-saturated sodium bicarbonate aqueous solution, and extracted with EtOAc (2x) and DCM (1x). The combined organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-60% 3: 1 EtOAc-EtOH / heptane), providing 4- (6- fluoro-7- (2- fluoro-6-hydroxyphenyl) -1- (2- isopropyl -4- methylpyridin-3-yl) -2- oxygen -1,2- dihydropyrido [2,3-d] pyrimidin-4-yl) -3- methylpiperazine -1- carboxylic acid (3S) - tert-butyl ester. 1 H NMR(400MHz,DMSO-d6)δppm 10.19(brs,1H),8.38(d,J=5.0Hz,1H),8.26(dd,J=12.5,9.2Hz,1H),7.23-7.28(m,1H),7.1 8(d,J=5.0Hz,1H),6.72(d,J=8.0Hz,1H),6.68(t,J=8.9Hz,1H),4.77-4.98(m,1H),4.24(br t,J=14.2Hz,1H),3.93-4.08(m,1H),3.84(br d,J=12.9Hz,1H),3.52-3.75(m,1H),3.07-3.28(m,1H),2.62-2.74(m,1H),1.86-1.93(m,3H),1.43-1.48(m,9 H), 1.35 (dd, J=10.8, 6.8Hz, 3H), 1.26-1.32 (m, 1H), 1.07 (dd, J=6.6, 1.7Hz, 3H), 0.93 (dd, J=6.6, 2.1Hz, 3H). 19F NMR (376MHz, DMSO-d6) δ: -115.65 (s, 1F), -128.62 (s, 1F). m / z (ESI, +ve ion): 607.3 (M+H) + .

[0321] Step 7: 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-acryloyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one. Trifluoroacetic acid (25 mL, 324 mmol) was added to a solution of (3S)-tert-butyl 4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (6.3 g, 10.4 mmol) in DCM (30 mL). The resulting mixture was stirred at room temperature for 1 h and then concentrated. The residue was dissolved in DCM (30 mL), cooled to 0 ° C, and treated with DIPEA (7.3 mL, 41.7 mmol) and acryloyl chloride (0.849 mL, 10.4 mmol) in DCM (3 mL; added dropwise by syringe). The reaction was stirred at 0 ° C for 10 min, then quenched with half-saturated aqueous sodium bicarbonate solution and extracted with DCM (2x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-100% 3: 1 EtOAc-EtOH / heptane) to provide 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propane)-3-pyridyl)-4-((2S)-2-methyl-4-(2-acryloyl)-1-piperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one. 1H NMR(400MHz,DMSO-d6)δppm 10.20(s,1H),8.39(d,J=4.8Hz,1H),8.24-8.34(m,1H),7.23-7.32(m,1H),7.19(d,J=5.0H z,1H),6.87(td,J=16.3,11.0Hz,1H),6.74(d,J=8.6Hz,1H),6.69(t,J=8.6Hz,1H),6.21(br d,J=16.2Hz,1H),5.74-5.80(m,1H),4.91(br s,1H),4.23-4.45(m,2H),3.97-4.21(m,1H),3.44-3.79(m,2H),3.11-3.31(m,1H),2.67-2. 77(m,1H),1.91(s,3H),1.35(d,J=6.8Hz,3H),1.08(d,J=6.6Hz,3H),0.94(d,J=6.8Hz,3H). 19 F NMR (376MHz, DMSO-d6) δppm -115.64 (s, 1F), -128.63 (s, 1F). m / z (ESI, +ve ion): 561.2 (M+H) + .

[0322] The present invention comprises the following steps, wherein resolution of the racemic-diketone in steps 4 and 5 facilitates successful separation of atropisomers:

[0323]

[0324] Representative process description of the present invention

[0325] Step 1

[0326]

[0327]

[0328] To a solution of 2,6-dichloro-5-fluoro-3-pyridinecarboxylic acid (Compound 1) (25 kg; 119.1 mol) in dichloromethane (167 kg) and DMF (592 g) was added oxalyl chloride (18.9 kg; 148.9 mol) while maintaining the internal temperature between 15-20°C. Additional dichloromethane (33 kg) was added as a rinse and the reaction mixture was stirred for 2 h. The reaction mixture was cooled and then quenched with ammonium hydroxide (40.2 L; 595.5 mol) while maintaining the internal temperature at 0±10°C. The resulting slurry was stirred for 90 min and the product was then collected by filtration. The filtered solid was washed with deionized water (3X 87 L) and dried to provide 2,6-dichloro-5-fluoronicotinamide (Compound 2).

[0329] Step 2

[0330]

[0331]

[0332] In Reactor A, oxalyl chloride (11.9 kg; 93.8 mol) was added to a solution of 2,6-dichloro-5-fluoronicotinamide (Compound 2) (16.27 kg; 77.8 mol) in dichloromethane (359.5 kg) while maintaining the temperature ≤ 25°C for 75 min. The resulting solution was then heated to 40°C ± 3°C and aged for 3 h. Using vacuum, the solution was distilled to remove dichloromethane until the solution was below the stirrer. Dichloromethane (300 kg) was then added and the mixture was cooled to 0 ± 5°C. To a clean, dry reactor (Reactor B), 2-isopropyl-4-methylpyridin-3-amine (aniline) (12.9 kg; 85.9 mol) was added, followed by dichloromethane (102.6 kg). The aniline solution was azeotropically dried by vacuum distillation while maintaining the internal temperature between 20-25°C, replaced with additional dichloromethane until the solution was dry by KF analysis (limit ≤ 0.05%). The solution volume was adjusted to approximately 23 L volume with dichloromethane. The dried aniline solution was then added to Reactor A while maintaining the internal temperature at 0±5°C throughout the addition. The mixture was then heated to 23°C and aged for 1 h. The solution was finely filtered into a clean reactor to give a DCM solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (Compound 3) and used directly in the next step.

[0333] Step 3

[0334]

[0335]

[0336] A solution of 2,6-dichloro-5-fluoro-N-{[4-methyl-2-(propan-2-yl)pyridin-3-yl]carbamoyl}pyridine-3-carboxamide (UREA (Compound 3)) (containing 15 kg; 38.9 mol) in dichloromethane was solvent exchanged to 2-MeTHF using vacuum distillation while maintaining the internal temperature at 20-25°C. The reactor volume was adjusted to 40 L and then charged with additional 2-MeTHF (105.4 kg). Sodium tert-butoxide (9.4 kg; 97.8 mol) was added while maintaining 5-10°C. The contents were warmed to 23°C and stirred for 3 h. The contents were then cooled to 0-5°C and a solution of ammonium chloride (23.0 kg; 430 mol) in 60 L of deionized water was added. The mixture was warmed to 20°C and deionized water (15 L) was added and aged for a further 30 min. Stirring was stopped and the layers separated. The aqueous layer was removed and deionized water (81.7 L) was added to the organic layer. A mixture of concentrated HCl (1.5 kg) and water (9 L) was prepared and then slowly added to the reactor until the measured pH was between 4 and 5. The layers were separated and the aqueous layer was back-extracted with 2-MeTHF (42.2 kg). The two organic layers were combined and washed with 10% citric acid solution (75 kg), followed by a mixture of water (81.7 L) and saturated NaCl (19.8 kg). The organic layer was then washed with saturated sodium bicarbonate (75 kg), and if necessary, repeated to achieve an aqueous solution target Ph ≥ 7.0. The organic layer was washed again with brine (54.7 kg) and then dried with magnesium sulfate (5 kg). The mixture was filtered to remove the magnesium sulfate and the filter bed was rinsed with 2-MeTHF (49.2 kg). The combined filtrate and washings were distilled to a volume of 40 L under vacuum. The concentrated solution was heated to 55 ° C, and heptane (10-12 kg) was slowly added until the cloud point was reached. The solution was cooled to 23 ° C over 2 h, and then heptane (27.3 kg) was added over 2 h. The product slurry was aged at 20-25 ° C for 3 h, then filtered and washed with a mixture of 2-MeTHF (2.8 kg) and heptane (9 kg). The product was dried using nitrogen and vacuum to give solid 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (racemic-diketone (Compound 4)).

[0337] Step 4

[0338]

[0339]

[0340]

[0341] Under a nitrogen atmosphere, in a container, (+)-2,3-dibenzoyl-D-tartaric acid (2.0 equiv.) was added to a stirred suspension of compound 4 (1.0 equiv.) in 2-methyltetrahydrofuran (7.0 L / kg). 2-MeTHF is chiral, but is used as a racemic mixture. Different enantiomers of 2-MeTHF were randomly incorporated into the co-crystal. The resulting suspension was warmed to 75°C and aged at 75°C until complete dissolution was observed (≤30 min). The resulting solution was finely filtered at 75°C into a second container. At a rate that maintained the internal temperature above 65°C, n-heptane (2.0 L / kg) was charged into the finely filtered solution. The solution was then cooled to 60°C, seeded with crystals (0.01 kg / kg), and aged for 30 minutes. The resulting suspension was cooled to 20°C over 4 hours and then sampled for chiral purity analysis by HPLC. The suspension was charged with n-heptane (3.0 L / kg) and then aged for 4 hours at 20°C under a nitrogen atmosphere. The suspension was filtered and the separated solid was washed twice with (2:1) n-heptane:2-methyltetrahydrofuran (3.0 L / kg). The material was dried with nitrogen and vacuum to obtain a meta-diketone:DBTA:Me-THF complex (Compound 4a).

[0342] Step 5

[0343]

[0344]

[0345] In container A, a suspension of disodium hydrogen phosphate (21.1 kg, 2.0 equivalents) in deionized water (296.8 L, 6.3 L / kg) was stirred until dissolution was observed (≥30 min). In container B, a suspension of dione: DBTA: Me-THF complex (composition 4a) [46.9 kg (corrected for dione as 25.9 kg, 1.0 equivalents)] in methyl tert-butyl ether (517.8 L, 11.0 L / kg) was stirred for 15 to 30 minutes. The resulting solution from container A was added to container B, and the mixture was then stirred for more than 3 hours. Stirring was stopped and the biphasic mixture was separated for more than 30 minutes. The aqueous phase below was removed and then stripped with methyl tert-butyl ether (77.7 L, 1.7 L / kg). The organic phases were combined in container B and dried with magnesium sulfate (24.8 kg, 0.529 kg / kg). The resulting suspension from container B was stirred for more than three hours and then filtered into container C. A rinse of methyl tert-butyl ether (46.9 L, 1.0 L / kg) was charged to container B and then filtered into container C. The contents of container C were cooled to 10°C and then distilled under vacuum while slowly warming to 35°C. Distillation was continued until 320-350 kg (6.8-7.5 kg / kg) of methyl tert-butyl ether were collected. After the contents of container C were cooled to 20°C, n-heptane (278.7 L, 5.9 L / kg) was charged over 1 hour and then distilled under vacuum while slowly warming to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane were collected. After the contents of container C were cooled to 20°C, n-heptane (278.7 L, 5.9 L / kg) was charged for the second time over 1 hour, and then distilled under vacuum while slowly warming to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg / kg) of a mixture of methyl tert-butyl ether and n-heptane was collected. After the contents of container C were cooled to 20°C, n-heptane (195.9 L, 4.2 L / kg) was charged for the third time over 1 hour, and then sampled for analysis of solvent composition by GC. The suspension in container C was continued to be stirred for more than one hour. The suspension was filtered and then washed from container C with n-heptane (68.6 L, 1.5 L / kg) rinse. The separated solid was dried at 50°C and a sample was submitted for storage. 7-Chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (meta-diketone) compound 5M was obtained.

[0346] The first generation process highlighted above has been successfully scaled up to 200+kg of racemic-dione starting material (Compound 5). In this process, inoculation with a thermodynamically stable racemic-dione crystalline form (which exhibits low solubility) resulted in batch failure. Based on our subsequent studies, we found that the robustness of the process was improved by adjusting the heptane feed schedule to increase the DBTA equivalent and reduce the seed temperature. The improved process can resist the presence of thermodynamically stable racemic-dione crystalline forms and promote the successful separation of atropisomers. Subsequent batches will incorporate the improved process for large-scale manufacturing.

[0347] Step 6

[0348]

[0349]

[0350]

[0351] 7-Chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (meta-diketone) (3.7 kg; 9.8 mol) was combined with 10.5 kg toluene in reactor (A) and distilled to oil to remove water while maintaining a set point of 45°C. Toluene (21 kg) was added to the residue and the mixture was stirred at 40-45°C for 30 min. The contents were cooled to 22°C and phosphorus oxychloride (1.8 kg; 11.7 mol) was then added. The mixture was cooled to 0-5°C and N,N-diisopropylethylamine (2.5 kg; 19.34 mol) was then added while maintaining the temperature <5°C. The solution was aged at 22°C for 3 h. In a separate reactor (B), (s)-1-boc-3-methylpiperazine (2.21 kg; 10.8 mol) and N,N-diisopropylethylamine (1.26 kg; 9.75 mol) were combined in toluene (6 kg) and then charged to reactor (A) while maintaining <25°C. The reaction mixture was aged at 22°C for 15 min and then quenched with a solution of sodium bicarbonate (973 g) in water (12.9 L) while maintaining the temperature <25°C. The mixture was stirred for 30 min and then DCM (36.8 kg) was added while stirring was continued for 1 h. The layers were allowed to separate and the lower organic layer was drained to reactor (C). The aqueous layer in reactor (A) was stripped with DCM (18.4 kg) and the combined organic layers were washed with brine solution (6.0 kg NaCl; 16.5 kg deionized water). The organic layer was distilled at atmospheric pressure, maintaining the internal temperature between 45-55°C. DCM was substituted with azeotropic drying solution during distillation. After distillation, DCM was used to adjust the solution volume to 19L. The solution was cooled to 30°C and finely filtered. The filtrate was merged with ethyl acetate (8.5kg) and then distilled at atmospheric pressure until 11-13kg was collected in the receiver. The solution was inoculated with 30g of the genuine product, then aged for 1h at 25-30°C, and then further distilled at atmospheric pressure at 45-55°C internal temperature until 8.2kg of distillate was collected. The slurry was cooled to 22°C and aged overnight, and then further cooled to 0-5°C. The product was collected by filtration and washed twice with ethyl acetate (4.2kg each time). The filter cake was dried with nitrogen and vacuum to give (3S)-tert-butyl 4-{7-chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate (Compound 6, PIPAZOLINE).

[0352] Step 7

[0353]

[0354]

[0355] Degassed dioxane (74.2 kg), (3S)-tert-butyl 4-{7-chloro-6-fluoro-(1M)-1-[4-methyl-2-(propane-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate (Compound 6, Pipazoline) (24.0 kg, 45.2 mol), potassium acetate (22.2 kg, 45.2 mol) and (dppf)PdCl2 (0.74 kg, 1.01 mol) were added to the reactor. The reactor was inertized with nitrogen. Nitrogen was bubbled into the solution until the oxygen content was <500 mg / L. The reaction was heated to 87.5°C. A solution of potassium trifluoro(2-fluoro-6-hydroxyphenyl)borate (12.6 kg, 54.3 moL) in degassed dioxane (49.4 kg) and degassed water (14.4 kg) with an oxygen content of <500 mg / L was transferred to the reaction, maintaining an internal temperature of 82.5°C ± 7.5°C. The reaction was adjusted to 87.5°C ± 1.5°C and stirred for 75 min ± 15 min. The reactor was charged with 1.0 M EDTA solution (47.3 kg) followed by water (40.1 kg) while maintaining an internal temperature of 85°C ± 5°C. The reaction mixture was cooled to 20°C ± 3°C over >2 h and then stirred for >16 h. The reaction was filtered and the crude solid was rinsed with water (3 x 120 kg). The solid was rinsed with a mixture of heptane (28.8 kg) and 2-propanol (33.1 kg) and then dried at <50°C for >10 h. A clean reactor was charged with the crude solid and dichloromethane (240 kg). The contents were stirred at 20°C ± 5°C for > 30 min. Silicon mercaptan (144 kg) and dichloromethane (14.9 kg) were added to the reactor. The reaction was stirred at 20°C ± 5°C for 18 h. The reaction was filtered and rinsed with dichloromethane (84 kg). The solution was distilled and the solvent was exchanged with 2-propanol. The reaction was heated to 60°C ± 3°C and heptane (108 kg) was charged while maintaining the reaction temperature at 60°C ± 3°C. The reaction was stirred for 45 min, then cooled and stirred at 20°C ± 5°C for 2.5 h. The reaction was filtered and rinsed with 50% by volume heptane / 2-propanol (61.9 kg). The isolated solid was dried at <50 °C for >12 h to give (3S)-tert-butyl 4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate (Compound 7, BIARYL).

[0356] Step 8

[0357]

[0358] General Note: All equivalents and volumes refer to BIARYL 7 report

[0359]

[0360] To the reactor was added (3S)-tert-butyl 4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propane-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate (Compound 7, BIARYL) (2.75 kg, 5.27 mol), DCM (13.7 L) and TFA (5.67 kg, 49.7 mol). The reaction was stirred at 20±5°C for 8-16 h. Potassium carbonate (11.24 kg), water (54.8 L) and methanol (13.7 L) were added to a second reactor to form a homogeneous solution. The reaction mixture was added to the potassium carbonate solution over 2 h. The mixture was stirred at 20±5°C for an additional 12 h. The resulting slurry was filtered and rinsed with water (2×27.5 L). The wet cake was dried for 24 h to give 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperazin-1-yl]-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (Compound 8, DESBOC).

[0361] Step 9

[0362]

[0363] General Note: All equivalents and volumes refer to Des-BOC reports

[0364]

[0365] 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperazin-1-yl]-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (Compound 8, DESBOC) (156.25 g) was combined with N-methylpyrrolidone (625 mL) and stirred at ambient temperature. Acryloyl chloride (36.29 g; 401.0 mmol) was added to the resulting solution.

[0366] While maintaining <30°C internal temperature. Stir the contents at 25°C for 2h. In a separate reactor, prepare a solution of disodium phosphate (175.1 g; 1234 mmol) in deionized water (3.1 L). The crude product solution is then transferred to the reactor containing the disodium phosphate solution at 25°C for >2h. The slurry is heated to 45°C when added halfway and aged at the same temperature for 2h after complete addition. The mixture is cooled to 25°C and aged for 4h, then the solid is collected by vacuum filtration. The solid was washed twice with water (1.5 L each time), and the product was dried under nitrogen and vacuum to give the product 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (crude compound 9). 4

[0367] Step 10

[0368]

[0369] General Note: All equivalents and volumes refer to the Crude Drug Substance Report

[0370]

[0371] 6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propane-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (crude compound 9) (142.33 g; 253.9 mmol) was combined with ethanol (996 mL) and water (270 mL). Acetic acid (21.8 ml; 380.8 mmoL) was added and the mixture was heated to 75°C to form a solution which was finely filtered into a clean reactor. The solution was cooled to 45°C and then water (1067 mL) was added while maintaining the internal temperature>40°C. The solution was seeded with authentic compound 9 and the resulting mixture was aged for 30 min. Water (1138 mL) was then added over 2 h. The mixture was cooled to 25 °C and aged for 8 h, then the solid was collected by vacuum filtration and washed with a mixture of ethanol (355.8 mL) and water (711.6 mL). The solid was dried using vacuum and nitrogen to afford 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propane-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (Compound 9).

[0372] Step A1 Reaction Scheme and Feed Table

[0373]

[0374]

[0375]

[0376] Reactor A was charged with THF (6 volumes) and diisopropylamine (1.4 equiv). The resulting solution was cooled to -70°C and n-BuLi (2.5M hexanes, 1.5 equiv) was slowly added. After the addition was complete, a solution of 3-fluoroanisole (1.0 equiv) in THF (6 volumes) was slowly added and kept at -70°C for 5 min. B(EtO)3 (2.0 equiv) was slowly added and kept at -70°C for 10 min. The reaction mixture was quenched with 2N HCl. The quenched reaction mixture was extracted with MTBE (3×4 volumes). The combined organic phases were concentrated to 1.5-3 total volumes. Heptane (7-9 volumes) was added dropwise and the mixture was cooled to 0-10°C and stirred for 3h. The mixture was filtered and rinsed with heptane (1.5 volumes). The solid was dried under nitrogen at <30°C to give (2-fluoro-6-methoxyphenyl)boronic acid.

[0377] Step A2 Reaction Scheme and Feed Table

[0378]

[0379]

[0380] Reactor A was charged with dichloromethane (4 volumes) and 2-fluoro-6-methoxy-4-methylphenylboronic acid (1 equivalent). The reaction mixture was cooled to -30 ° C, and 1.5BBr3 (1.5 equivalents) was added dropwise. After addition, the mixture was warmed to 25 ° C and stirred for 2 h. The reaction mixture was quenched in ice-cold (0-5 ° C) water (10 volumes). MTBE (10 volumes) was added, the mixture was warmed to 25 ° C, and stirred for 1-2 h or until all solids dissolved. The aqueous phase was separated and extracted with MTBE (3 volumes). The combined organic extracts were washed with water (3 volumes) and then concentrated to 1 total volume. Heptane (10 volumes) was added to the mixture and stirred for 2 h. The resulting product was separated by filtration and dried at <30 ° C to obtain (2-fluoro-6-hydroxyphenyl) boronic acid.

[0381] Step A3 Reaction Scheme and Feed Table

[0382]

[0383]

[0384] Step A3

[0385] Potassium fluoride (21.0 kg; 20.87 mol) was combined with water (28 L) in a reactor (reactor A) and the contents were stirred for 30 min. In a separate reactor (reactor B), (2-fluoro-6-hydroxyphenyl)boric acid (14.00 kg, 89.79 mol) was charged at 25°C, followed by acetonitrile (206.1 kg) and citric acid (30.94 kg; 147.26 mol). The contents of reactor A were added to reactor B at 25°C and stirred at that temperature for 10 h. The reaction mixture was filtered through a diatomaceous earth bed (7.0 kg) and rinsed with acetonitrile (42 kg). The filtrate was combined with isopropanol (56 kg) and then distilled under vacuum at a temperature of <35°C, replacing the distillation volume of the reactor with isopropanol, and repeated as needed to complete the solvent exchange from acetonitrile to isopropanol. The slurry was cooled to 15°C and aged for 1 h, then filtered and washed with 28 kg of isopropanol. The filter cake was dried using vacuum and nitrogen and packaged to give Compound A3.

[0386] Resolution of diketone compound 5

[0387] Chromatographic separation of diketone intermediates

[0388] A number of chiral chromatography techniques and methods were used to separate the meta-diketone from compound 4. The techniques and stationary phases are well known in the art and are listed in Table 1.

[0389]

[0390] Table 1

[0391]

[0392] ^Yield is defined as the % of available meta-diketone recovered in the desired purity of >98% ee.

[0393] *This separation was performed multiple times. For each batch of material, the mobile phase may have been modified slightly to accommodate variations within the batch. Other mobile phases used for the purification include:

[0394] 1) 25 / 75 methanol / CO2,

[0395] 2) 30 / 70 methanol / CO2, and

[0396] 3) 50 / 50 methanol / CO2.

[0397] SFC, HPLC and SMB techniques are well known in the art and Stationary phases are available from commercial sources such as Fisher Scientific and Daicel Corporation.

[0398] However, it is desirable to develop more efficient methods for isolating meta-diketones (compound 5).

[0399] Classic Split

[0400] The present invention relates to the development of a viable classical resolution method for the meta / para-diketone racemate (compound 4).

[0401] A total of 100 cocrystal screening experiments were performed, and three potential diketone cocrystals were identified. (+)-2,3-Dibenzoyl-D-tartaric acid (DBTA) was selected as the chiral reagent for resolution based on the highest area ratio of meta / para-diketone in the residual solid and the lowest area ratio in the supernatant.

[0402] Based on the results of 100 co-crystal screening experiments and 20 more solvent screenings, it was found that 2-MeTHF / n-heptane provided better resolution results than other solvent systems. Based on the solubility results of the meta-diketone co-crystal and the para-diketone co-crystal in different ratios of 2-MeTHF and n-heptane, 2-MeTHF / n-heptane (1.4:1, v / v) was selected as the optimal solvent composition for resolution.

[0403] In order to find out any possible forms that were converted to diketone racemates or meta / para-diketones during the chiral resolution crystallization process, the solubility of meta-diketone co-crystals, para-diketone co-crystals, meta+para-diketone co-crystal mixtures (1:1, w / w), diketone racemates, and DBTA in 2-MeTHF / n-heptane (1.4:1, v / v) at different temperatures was determined. No form change of meta-diketone co-crystals and para-diketone co-crystals was observed for 7 days at different temperatures. However, after stirring the mixture of meta+para-diketone co-crystal mixtures (1:1, w / w) at different temperatures, the C-type diketone racemate was obtained. After stirring the diketone racemates at the corresponding temperatures for 7 days, the D-type diketone racemate (20 and 30°C) or the C-type diketone racemate (40, 50, 60, and 65°C) was observed. At all temperatures, the solubility of DBTA was observed to be about 100 mg / mL.

[0404] To further optimize the resolution process, a ternary phase diagram of the meta / para-diketone cocrystal was drawn based on the equilibrium solubility results, and no eutectic point was obtained, which may be because when the meta-diketone cocrystal and the para-diketone cocrystal are present, the C-type racemate will crystallize out. Another ternary phase diagram of the meta / para-diketone was drawn based on the equilibrium solubility results, and no eutectic point was obtained, which may be because when the meta-diketone cocrystal and the para-diketone are present, the C-type or D-type diketone racemate will crystallize out.

[0405] In summary, a chiral reagent (DBTA) and solvent system ((2-MeTHF / n-heptane (1.4:1, v / v)) for the resolution of dione racemates have been identified. Using this resolving agent and solvent system, a small-scale crystallization process can achieve a yield of 39% and an ee purity of 99% for the meta-diketone. In addition, the polymorphism of the dione racemate was observed and studied in the screening experiment.

[0406] 2 Screening experiments

[0407] 2.1 Cocrystal screening

[0408] A total of 100 co-crystal screening experiments were performed using 20 acids and 5 solvent systems (the results are summarized in Table 2-1). Typically, the diketone racemate and acid were mixed at a molar ratio of 1:1 and stirred at room temperature for 3 days before being separated for XRPD. Based on the XRPD results, three potential acids that could form co-crystals of the diketone racemate were identified, including (1S)-(-)-camphoric acid ( Figure 2-2 ), (+)-2,3-dibenzoyl-D-tartaric acid ( Figure 2-3 ) and D-(+)-malic acid ( Figure 2-4 Based on the XRPD results, four new free base crystalline forms were also obtained and designated as Type B-E diketone racemates.

[0409] As shown in Table 2-2, the supernatant and residual solid of three potential co-crystals were further tested by HPLC. The ratio of meta-diketone / para-diketone was measured and summarized in Table 2-2. As a result, the DBTA co-crystal showed an area ratio of meta-diketone / para-diketone of 0.11 in the supernatant and 4.4 in the residual solid, indicating that meta-diketone and para-diketone showed good resolution after forming co-crystals with DBTA. Therefore, DBTA was selected as a chiral reagent for further resolution optimization.

[0410] Table 2-1 Summary of co-crystal screening experiments

[0411]

[0412]

[0413]

[0414] + represents the crystal form of the free racemic-dione (no cocrystal is formed), *: (1S)-(-)-camphoric acid cocrystal, # :(+)-2,3-Dibenzoyl-D-tartaric acid cocrystal, $ :D-(+)-Malic acid eutectic

[0415] Table 2-2 Summary of HPLC data of three co-crystals

[0416]

[0417] 2.2 Solvent screening

[0418] In order to select suitable solvent to further split meta-diketone and para-diketone, the area ratio of HPLC of meta-diketone in more than 20 solvents / solvent mixtures was collected. As listed in Table 2-3, 2-MeTHF shows the best splitting, wherein the meta-diketone / para-diketone area ratio is 0.7 in the supernatant and 4.1 in the residual solid. However, 2-MeTHF / normal-heptane (1:1, v / v) shows better splitting results (Table 2-2) during the co-crystal screening process, so 2-MeTHF / normal-heptane is selected for further optimization. The area ratio of HPLC of meta-diketone is collected with different acid / base ratios in 2-MeTHF / normal-heptane of different ratios. The results in Table 2-4 show that a higher acid / FB (2:1 or 1.5:1) ratio is required in 2-MeTHF / normal-heptane (8:1 or 4:1, v / v) to improve the ratio of meta-diketone in the separated solid.

[0419] The solubility of the meta-cocrystals and para-cocrystals in different ratios of 2-MeTHF / n-heptane was also conducted at 5°C and 25°C, which are summarized in Table 2-5. The meta-cocrystals were provided by the customer, and the para-cocrystals were prepared by reverse anti-solvent and anti-solvent (see Section 4.3 for experimental details). The solubility results in Table 2-5 show that a volume ratio of 2-MeTHF / n-heptane of 1.5:1 can obtain the best resolution at room temperature. The customer conducted more resolution experiments, among which a volume ratio of 1.4:1 showed the best resolution results. Therefore, a volume ratio of 2-MeTHF / n-heptane of 1.4:1 was selected as the solvent system for resolution.

[0420] Table 2-3 Solvent screening of diketone racemic DBTA cocrystals (area ratio of meta-diketone / para-diketone)

[0421]

[0422] NA: A clear solution was obtained and no solids were isolated.

[0423] Table 2-4 Results of screening the acid / base ratio and 2-MeTHF / n-heptane ratio using the diketone racemic DBTA cocrystal (meta-diketone / para-diketone area ratio)

[0424]

[0425]

[0426] L: supernatant, S: solid, CA: Form A cocrystal, DA: Form A diketone racemate

[0427] Table 2-5 Screening of 2-MeTHF / n-heptane ratios for m / p-diketone cocrystals

[0428]

[0429] *: Type A eutectic, # :A-type eutectic

[0430] 2.3 Solubility of diketone DBTA cocrystals, diketone racemates, and DBTA

[0431] The 7-day equilibrium solubility of the meta-diketone cocrystal, para-diketone cocrystal, meta+para-diketone cocrystal mixture (1:1, w / w) and diketone racemate was established in 2-MeTHF / n-heptane (1.4:1, v / v) at different temperatures (20°C, 30°C, 40°C, 50°C, 60°C, 65°C, 75°C and 80°C). A color change was observed after 5 days at 75°C and 80°C, indicating degradation, so no solubility was collected.

[0432] When the meta-cocrystals and para-cocrystals were stirred at different temperatures for 7 days, no form change was observed ( Figure 2-5 and Figure 2-6 After stirring the mixture of the meta-diketone cocrystal and the para-diketone cocrystal mixture (1:1, w / w) at different temperatures for 7 days, the C-type diketone racemate ( Figure 2-7 After stirring the diketone racemate at different temperatures for 7 days, the D-type diketone racemate (20°C and 30°C) and the C-type diketone racemate (40°C, 50°C, 60°C and 65°C) were observed ( Figure 2-8 and Figure 2-9 ).

[0433] The 5-day equilibrium solubility of DBTA was established in 2-MeTHF / n-heptane (1.4:1, v / v) at different temperatures (20°C, 30°C, 40°C, 50°C, 60°C and 65°C). A solubility of approximately 100 mg / mL was observed at all temperatures. No significant differences were observed at different temperatures (Tables 2-7).

[0434] Table 2-6 Solubility of diketone DBTA cocrystal, mixture of meta / para-diketone cocrystal, and diketone racemate in 2-MeTHF / n-heptane (1.4:1, v / v)

[0435]

[0436] Table 2-7 Solubility of DBTA in 2-MeTHF / n-heptane (1.4:1, v / v)

[0437] Sample ID Temperature(℃) Solubility (mg / mL) 1-15-A1 20 99.1 1-15-A2 30 100.3 1-15-A3 40 98.9 1-15-A4 50 88.0 1-15-A5 60 105.0 1-15-A6 65 96.1

[0438] 2.4 Ternary phase diagram

[0439] 2.4.1 m / p-diketone eutectics

[0440] The meta-diketone co-crystal and the para-diketone co-crystal were weighed, the corresponding masses are listed in Table 2-8, and stirred in 2-MeTHF / n-heptane (1.4:1, v / v) at room temperature for 72 hours. A ternary phase diagram of the meta / para-diketone co-crystal was drawn based on the 72-hour equilibrium solubility data, and no eutectic point was obtained ( Figure 2-10 ).

[0441] Table 2-8 Summary of solubility data of m / p-diketone cocrystals

[0442]

[0443] 2.4.2 m / p-dione

[0444] Weigh the meta-diketone and para-diketone, the corresponding masses are listed in Table 2-9, and stir for 5 days in 2-MeTHF / normal heptane (1.4: 1, v / v) at room temperature. Draw a ternary phase diagram based on 5 days of equilibrium solubility data in 1.0 mL 2-MeTHF / normal heptane (1.4: 1, v / v) at room temperature. Observe the A-type meta-diketone, A-type para-diketone, C-type and D-type diketone racemates in the residual solid of the solubility sample. No eutectic point is obtained in the phase diagram ( Figure 2-11 ).

[0445] Table 2-9 Data summary of the ternary phase diagram of m / p-diketone

[0446]

[0447]

[0448] RC: C-type diketone racemate; RD: D-type diketone racemate; A: A-type meta- or para-diketone (the XRPD patterns of A-type meta-diketone and A-type para-diketone are identical and indistinguishable).

[0449] 3 Solid-state characterization of crystalline forms

[0450] A total of five diketone racemic crystal forms and two cocrystal forms were obtained. All of these forms were characterized by XRPD, TGA, DSC, PLM and 1 H NMR characterization and are summarized in Tables 2-10. The solid state characterization data indicated that the diketone racemates Forms A and D were identified as 2-MeTHF solvates, Form B was identified as an acetone solvate, Form C was identified as an anhydrate, and Form E was identified as an MTBE solvate.

[0451] It was found that both the A-type meta-diketone cocrystal and the A-type para-diketone cocrystal were 2-MeTHF solvates. All characterization data are shown in Figure 3-1 to Figure 3-22 middle.

[0452] Table 2-10 Summary of crystal forms

[0453]

[0454]

[0455] *: exothermic peak; &: not detected.

[0456] 3.1.1 Competitive slurry of diketone racemate form

[0457] The B-E diketone racemates were successfully reproduced at room temperature by slurrying the A-type diketone racemate in acetone, H2O / ACN (1:1, v / v), 2-MeTHF / n-heptane (1.4:1, v / v) and MTBE / n-heptane (1:1, v / v), respectively.

[0458] About 5 mg of each diketone racemic form (Forms A to E) was weighed into an HPLC vial, 0.3 mL of a saturated solution of the diketone racemate in 2-MeHTF / n-heptane (1.4:1, v / v) was added to the vial, and the mixture was then stirred at 20°C, 30°C, 40°C, 50°C, 60°C and 65°C for 5 days.

[0459] All free base forms were converted to the C-diketone racemate by competitive slurry in 2-MeHTF / n-heptane (1.4:1, v / v) at the target temperature, indicating that the C-diketone racemate is the most thermodynamically stable form in 2-MeHTF / n-heptane (1.4:1, v / v) at 20°C to 65°C.

[0460] Table 2-11 Competitive slurry results

[0461]

[0462] 3.2 Preparation of diketone cocrystals

[0463] 3.2.1 Small scale

[0464] At 65°C, 2 g of p-diketone and 1 g of DBTA were dissolved in 18 mL of 2-MeTHF to give an almost clear solution. 18 mL of heptane was added to this solution over 1 h. The solution was cooled to 20°C over 4 h and aged overnight. The solution was evaporated at room temperature for about 1 h using air blowing, and a light yellow oily paste was obtained. Another 54 mL of heptane was added to the mixture, with stirring for 2 h. The suspension was filtered. The solid sample was designated as 810465-16-A.

[0465] 3.2.2 Large Scale

[0466] At 65 ℃, 10 g of p-diketone and 5 g of DBTA were dissolved in 100 mL of 2-MeTHF. The solution was filtered with a 0.45 μm PTFE filter and a clear solution was obtained. The clear solution was added dropwise to a suspension of 400 mL of heptane containing about 1 g of seed (810465-16-A) produced by the first run. The suspension was kept stirring for 5 h at room temperature and then separated. About 10 g of p-diketone cocrystal (810465-20-A) was produced with a yield of about 66%.

[0467] 4 Instruments and methods

[0468] 4.1 XRPD

[0469] For XRPD analysis, a PANalytical X-ray powder diffractometer was used in reflection mode. The XRPD parameters used are listed in Table 4-1.

[0470] Table 4-1 Parameters of XRPD test

[0471]

[0472]

[0473] 4.2 TGA and DSC

[0474] TGA data were collected using a TA Discovery 550, Q500 and Q5000 TGA from TA Instruments. DSC was performed using a Q500, Q5000 and Discovery 2500 DSC from TA Instruments. The detailed parameters used are listed in Table 4-2.

[0475] Table 4-2 Parameters of TGA and DSC tests

[0476] parameter TGA DSC method rise rise Sample tray Aluminum, Open Aluminum, Curled temperature RT-350℃ 25℃-300℃ Heating rate 10℃ / min 10℃ / min Purge gas <![CDATA[N2]]> <![CDATA[N2]]>

[0477] 4.3HPLC

[0478] The solubility was tested using Agilent 1100 / 1260 HPLC, and the detailed method is listed in Table 4-3.

[0479] Table 4-3 HPLC method for solubility testing

[0480]

[0481]

[0482] Table 4-4 HPLC method for solubility testing (DBTA)

[0483]

[0484] 4.4 1 H NMR

[0485] DMSO-d6 was used as solvent and the data were collected on a Bruker 400M NMR spectrometer. 1 H NMR spectrum.

[0486] 4.5PLM

[0487] Polarized light microscopy images were captured on a Nikon DS-Fi2 upright microscope at room temperature.

[0488] Compound 5 was additionally screened with 1,3-diphenyl-3-oxopropanesulfonic acid 11b.

[0489] Due to the low basicity of the pyridine moiety in compound 5 and limited 'hits' in forming crystalline salts using the standard screening setup, it was chosen to screen racemic compound 4 with 1,3-diphenyl-3-oxopropanesulfonic acid 11b at 0.06 mmol scale.

[0490]

[0491] Gram-scale resolution of racemic compound 5: A 250 mL round-bottom flask was charged with 2.0 g of racemic compound 4 (5.7 mmol, 1.0 equiv) in 200 mL of EtOH:AcOH (90:10 v:v). After the material had dissolved, 832 mg of sulfonic acid 11b (2.9 mmol, 0.5 equiv) was added to the solution. The clear solution was stirred at a stirring speed of 800 rpm for 15 hours. A white precipitate was formed, which was separated from the mother liquor. The isolated salt was suspended in CH2Cl2 and treated with concentrated aqueous NaHCO3 using a separatory funnel. The organic layer was separated and the alkaline aqueous layer was extracted with CH2Cl2 (2x). The organic layers were combined and dried over Na2SO3. The solvent was evaporated to give 415 mg of (m-)-5 (96% ee) (see Figure 6-1 ).

[0492] The clear mother liquor was evaporated to dryness. The yellow oily material was dissolved in CH2Cl2 and treated with concentrated aqueous NaHCO3 using a separatory funnel. The organic layer was separated and the basic aqueous layer was extracted with CH2Cl2 (2x). The organic layers were combined and dried over Na2SO3. Evaporation of the solvent gave 1579 mg of 5 (23% ee in the (para)-atropisomer; Figure 6-2 ).

[0493] Polymorph Screening of Meta-diketone DBTA Cocrystals

[0494] Characterization of the Crystalline Form of 5-Diketone DBTA Cocrystals

[0495] The polymorph screening experiment of meta-diketone was established under 100 conditions using slurry conversion, slow evaporation, slow cooling, anti-solvent addition, vapor diffusion, temperature cycling and wet milling. A total of 17 crystal forms (A to Q forms) were obtained from the screening. The form relationship is as follows Figure 4-1 Detailed characterization data are provided in Table 5-1 and Figure 5-1 An overlay of the XRPD patterns is shown in . Solid state characterization results indicated that Form G is a hydrate, while the other forms are solvates.

[0496] 5.1 Instruments and methods

[0497] 5.1.1 XRPD

[0498] Using Panalytical X'Pert 3 Powder XRPD XRPD was performed on a Si zero background support. The 2θ position was calibrated relative to a Panalytical Si reference standard disk. The parameters used are listed in Table 5-a.

[0499] Table 5-a Parameters used for XRPD testing

[0500]

[0501] 5.1.2TGA / DSC

[0502] TGA data were collected using a TA Discovery 550 TGA from TA Instruments. DSC was performed using a TAQ2000 DSC from TA Instruments. The DSC was calibrated with an indium reference standard, and the TGA was calibrated with a nickel reference standard. The detailed parameters used are listed in Table 5-b.

[0503] Table 5-b Parameters used for TGA and DSC tests

[0504]

[0505] 5.2 Polymorph screening

[0506] The solubility of Form A (3-05-A) was estimated at room temperature. About 2 mg of solid was added to a 3 mL glass vial. The solvent in Table 5-c was then added to the vial stepwise (50 μL / 50 μL / 200 μL / 700 μL) until the solid dissolved or the total volume reached 2 mL. The results summarized in Table 5-c were used to guide solvent selection in polymorph screening.

[0507] Polymorph screening experiments were performed using different crystallization or solid transformation methods. The methods used and the crystalline forms identified are summarized in Table 5-c.

[0508] Table 5-c Approximate solubility of starting material (6010013-05-A) at room temperature

[0509] Solvents Solubility (mg / mL) Solvents Solubility (mg / mL) MTBE S<3.1 acetone S>52.0 <![CDATA[H2O]]> 2.4<S<8.0 DMF S>40.0 n-Heptane 3.0<S<10.0 Anisole S>40.0 Toluene 2.7<S<9.0 Acetic acid S>40.0 Hexane 2.9<S<9.7 THF S>50.0 IPA 26.0<S<52.0 ACN S>46.0 2-MeTHF 24.0<S<48.0 CHCl3 S>50.0 1,4-Dioxane 25.0<S<50.0 EtOAc S>72.0 n-BuOH 23.0<S<46.0 DMSO S>72.0 MIBK 38.0<S<76.0 MeOH S>78.0 B O S>28.0 EtOH S>68.0 IPA S>32.0 - -

[0510] Table 5-d Summary of polymorph screening experiments

[0511]

[0512]

[0513] 5.2.1 Slurry at room temperature

[0514] Slurry experiments were performed in different solvent systems at room temperature. Approximately 20 mg of Form A (3-05-A) was suspended in 0.2 mL of solvent in a 3-mL glass vial. After the suspension was magnetically stirred at room temperature for 13 days, the remaining solid was separated for XRPD analysis. The results summarized in Table 5-e indicate that Forms A to D and Form J were obtained.

[0515] Table 5-e Summary of slurry experiments at room temperature

[0516]

[0517]

[0518] *: Solid obtained by slow evaporation at room temperature

[0519] 5.2.2 Slow evaporation

[0520] Slow evaporation experiments were performed under 16 conditions. Briefly, 20 mg of Form A (3-05-A) was dissolved in 0.2-0.8 mL of solvent in a 20 mL glass vial. If dissolution was not achieved, the suspension was filtered using PTFE (pore size 0.2 μm) and the filtrate was used in the following steps. Visually clear solutions were filtered with a 5-10 pinhole Cover and evaporate at room temperature. The isolated solid was used for XRPD analysis. The results summarized in Table 5-f show that Form A, Form C, Form D, Form J, Form K, Form L, Form N, Form O were obtained.

[0521] Table 5-f Summary of slow evaporation experiments

[0522]

[0523]

[0524] 5.2.3 Slow Cooling

[0525] Slow cooling experiments were performed in 9 solvent systems. About 20 mg of Form A (3-05-A) was suspended in 1 mL of solvent in a 3 mL glass vial at room temperature. The suspension was then heated to 50°C, balanced for two hours, and filtered using a PTFE membrane (pore size of 0.20 μm). The filtrate was slowly cooled to 5°C at a rate of 0.1°C / min. The results summarized in Table 5-g show that Form C, Form G, Form J, Form L, and Form O were observed.

[0526] Table 5-g Summary of slow cooling experiments

[0527]

[0528] *: Solid obtained from evaporation at room temperature.

[0529] 5.2.4 Antisolvent addition

[0530] A total of 9 anti-solvent addition experiments were performed. About 20 mg of the starting material (3-05-A) was dissolved in 0.2-1.4 mL of solvent to obtain a clear solution. The solution was stirred magnetically, and then 0.2 mL of anti-solvent was gradually added until precipitation occurred or the total amount of anti-solvent reached 15.0 mL. The precipitate obtained was separated for XRPD analysis. The results in Table 5-h show that Type A, Type C, Type H and Type I were obtained.

[0531] Table 5-h Summary of antisolvent addition experiments

[0532]

[0533] *: Solid obtained from evaporation at room temperature.

[0534] 5.2.5 Liquid vapor diffusion

[0535] Five liquid vapor diffusion experiments were carried out. About 20 mg of starting material (3-05-A) was dissolved in a suitable solvent to obtain a clear solution in a 3 mL vial. This solution was then placed in a 20 mL vial containing 3 mL of a volatile solvent. The 20 mL vial was sealed with a cap and kept at room temperature to allow enough time for organic vapor to interact with the solution. Separation precipitation was used for XRPD analysis. The results summarized in Table 5-i show that L-type, M-type and Q-type were produced.

[0536] Table 5-i Summary of liquid vapor diffusion experiments

[0537] Experiment ID Solvents Antisolvent Solid form 3-11-A1 MIBK n-Heptane Q-Type 3-11-A2 EtOAc IPA Type M 3-11-A3 THF MTBE L-type 3-11-A4 2-MeTHF n-Heptane L-type 3-11-A5* DMF Toluene gel

[0538] *: Solid obtained by evaporation at room temperature.

[0539] 5.2.6 Solid vapor diffusion

[0540] Solid vapor diffusion experiments were performed using 6 different solvents. About 10 mg of the starting material (3-05-A) was weighed and placed in a 3 mL vial, which was then placed in a 20 mL vial containing 2 mL of a volatile solvent. The 20 mL vial was sealed with a cap and kept at room temperature for 7 days to allow the solvent vapor to interact with the sample. The solid was tested by XRPD, and the results summarized in Table 5-j showed that Type A and Type M were produced.

[0541] Table 5-j Summary of solid vapor diffusion experiments

[0542] Experiment ID Solvents Solid form 3-12-A1 EtOH Type M 3-12-A2 MTBE Type A 3-12-A3 H2O Type A 3-12-A4 acetone Amorphous 3-12-A5 2-MeTHF Type A 3-12-A6 IPA Type A

[0543] 5.2.7 Temperature Cycle

[0544] Temperature cycle experiments were carried out in 7 solvent systems. At room temperature, about 20 mg of starting material (3-05-A) was suspended in 1 mL of solvent in a 3 mL glass vial. The suspension was then heated to 50 ° C, balanced for one hour, and filtered using a PTFE membrane (pore size of 0.20 μm). The filtrate was slowly cooled to 5 ° C at a rate of 0.2 ° C / min, and then heated to 50 ° C at a rate of 1 ° C / min. The cycle was repeated once more, and then cooled to 5 ° C at a rate of 0.2 ° C / min. Before separating the solid and analyzing these solids using XRPD, the sample was stored at 5 ° C. The results summarized in Table 5-k show that Type A, Type G and Type O are observed.

[0545] Table 5-k Summary of temperature cycling experiments

[0546] Experiment ID Solvent (v:v) Solid form 3-13-A1* 2-MeTHF Type A 3-13-A2* MeOH Type G 3-13-A3* MIBK Type A 3-13-A4* ACN Amorphous 3-13-A5 CHCl3 Low crystallinity 3-13-A6* Toluene Amorphous 3-13-A7* IPA Type O

[0547] *: Solid obtained from evaporation at room temperature.

[0548] 5.2.8 Slurry at 5°C

[0549] Slurry experiments were performed at 5°C in different solvent systems. Approximately 20 mg of the starting material (3-05-A) was suspended in 0.2 mL of solvent in a 3-mL glass vial. After the suspension was magnetically stirred at 5°C for 7 days, the remaining solid was separated for XRPD analysis. The results summarized in Table 5-1 show that Form A, Form C to Form E, and Form J were obtained.

[0550] Table 5-1 Summary of slurry experiments at 5°C

[0551] Experiment ID Solvent (v:v) Solid form 3-14-A1* B O Type C 3-14-A2* IPA Type D 3-14-A3* acetone Low crystallinity 3-14-A4* DMF gel 3-14-A5 Anisole Type E 3-14-A6* 2MeTHF Type A 3-14-A7* ACN Low crystallinity 3-14-A8* CHCl3 Low crystallinity 3-14-A9* EtOAc Low crystallinity 3-14-A10* MeOH Type C 3-14-A11* THF J-Type

[0552] *: Solid obtained from evaporation at room temperature.

[0553] 5.2.9 Wet grinding

[0554] Wet milling experiments were performed under five conditions. Briefly, 10 mg of Form A (3-05-A) was placed in a mortar and ground in approximately 20 μL of solvent for 5 min. The isolated solid was used for XRPD analysis. The results summarized in Table 5-m indicate that Form A was obtained.

[0555] Table 5-m Summary of wet grinding experiments

[0556]

[0557]

[0558] Table 5-1 Characterization of diketone DBTA cocrystal forms

[0559]

[0560]

[0561] Type 5.3A

[0562] Type A (3-05-A) is provided by the customer. Figure 5-4 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-5 As shown in the TGA and DSC data in , up to 125 ° C, the weight loss was 7.3%, and two endothermic peaks were observed at 109.4 and 120.0 ° C (peak). Figure 5-6 As shown, in 1 The presence of 2-MeTHF was confirmed in the H NMR spectrum. Based on the results, Form A was considered to be a 2-MeTHF solvate.

[0563] Type 5.4B

[0564] A sample of Form B (3-07-A1) was obtained by slurry of Form A in MTBE at room temperature. Figure 5-7 The XRPD pattern shown in Figure 2 indicates crystallinity. Figure 5-10 As shown in the TGA and DSC data in , up to 125 ° C, the weight loss was 7.2%, and an endothermic peak was observed at 115.7 ° C (peak). Figure 5-9 As shown, in 1 The presence of MTBE was confirmed in the H NMR spectrum. Based on the results, Form B is likely a MTBE solvate.

[0565] Type 5.5C

[0566] A sample of Form C (3-08-A5) was obtained by slow evaporation in EtOAc at room temperature. Figure 5-10 The XRPD pattern shown in indicated crystallinity. Figure 5-11 The TGA and DSC data shown in Figure 2 show that up to 125°C, there is a weight loss of 8.0% and two endothermic peaks at 92.7°C and 116.4°C (peak). Figure 5-12 As shown, in 1 The presence of EtOAc was confirmed in the H NMR spectrum. Based on the results, Form C is likely an EtOAc solvate.

[0567] Type 5.6D

[0568] Form D (3-07-A12) was obtained by slurry of Form A in IPAc at room temperature. Figure 5-13 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-14As shown in the TGA and DSC data in , up to 130°C, the weight loss was 7.5%, and there were endothermic peaks at 75.4°C, 110.5°C, 148.0°C and 116.6°C (peak), and an exothermic peak was observed at 265.9°C. Figure 5-15 As shown, in 1 The presence of IPAc was confirmed in the H NMR spectrum. Based on the results, form D is likely an IPAc solvate.

[0569] Type 5.7E

[0570] Form E (3-07-A15) was obtained by slurry of Form A in anisole at room temperature. Figure 5-16 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-17 As shown in the TGA and DSC data in , up to 125 ° C, the weight loss was 8.6%, and two endothermic peaks were observed at 103.8 ° C and 119.0 ° C (peak). Figure 5-18 As shown, the presence of IPAc was confirmed in the 1H NMR spectrum. Based on the results, Form E is likely to be anisole solvate.

[0571] 5.8F

[0572] Form F (3-07-A19) was obtained by slurry of Form A in IPAc / H2O (v:v 1:9) at room temperature. Figure 5-19 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-20 As shown in the TGA and DSC data in , up to 130 ° C, the weight loss was 6.2%, and two endothermic peaks were observed at 86.5 ° C and 107.9 ° C (peak). Figure 5-18 As shown, in 1 The presence of IPAc was confirmed in the H NMR spectrum. Based on the results, Form F is likely an IPAc solvate.

[0573] 5.9G

[0574] Form G (3-07-A26) was obtained by slurry of Form A in MeOH / H2O (aw=0.8) at room temperature. Figure 5-22 The XRPD results shown in the figure indicate a crystalline state. Figure 5-23 As shown in the TGA and DSC data in , up to 100°C, the weight loss was 6.4%, and endothermic peaks were observed at 86.0°C, 127.2°C, and 133.1°C (peak). Figure 5-24 As shown, in the solution 1 No MeOH or MeTHF signals were observed in the HNMR spectrum. Based on the results, Form G is likely a hydrate.

[0575] 5.10H

[0576] Form H (3-10-A1) was obtained by anti-solvent addition using acetone / H2O. Figure 5-25 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-26 As shown in the TGA and DSC data in , up to 130 ° C, the weight loss was 3.6%, and an endothermic peak was observed at 107.6 ° C (peak). Figure 5-27 As shown, in 1 The presence of acetone was confirmed in the H NMR spectrum. Based on the results, the H form is likely an acetone solvate.

[0577] 5.11I

[0578] Form I (3-10-A3) was obtained by anti-solvent addition using DMSO / H2O. Figure 5-28 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-29 As shown in the TGA and DSC data in , up to 150 ° C, the weight loss was 7.3%, and an endothermic peak was observed at 128.9 ° C (peak). Figure 5-30 As shown, in 1 The presence of DMSO was confirmed in the H NMR spectrum. Based on the results, Form I is likely a DMSO solvate.

[0579] 5.12J

[0580] Form J (3-08-A2) was obtained by slow evaporation in THF. Figure 5-31 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-32 As shown in the TGA and DSC data in , up to 125 ° C, the weight loss was 7.3%, and an endothermic peak was observed at 115.7 ° C (peak). Figure 5-33 As shown, in 1 The presence of THF was confirmed in the H NMR spectrum. Based on the results, Form J is likely a THF solvate.

[0581] 5.13K

[0582] Form K (3-08-A14) was obtained by slow evaporation in acetone. Figure 5-34 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-35 As shown in the TGA and DSC data in , up to 150°C, the weight loss was 5.7%, and endothermic peaks were observed at 96.7°C, 119.8°C, and 147.4°C (peak), and an exothermic peak was observed at 157.4°C (peak). Figure 5-36 As shown, in 1 The presence of acetone was confirmed in the H NMR spectrum. Based on the results, form K is likely an acetone solvate.

[0583] 5.14L

[0584] Form L (3-11-A4) was obtained by liquid vapor diffusion in 2-MeTHF / n-heptane. Figure 5-37 The XRPD results shown in show crystals with a preferred orientation. Figure 5-38 As shown in the TGA and DSC data in , up to 130 ° C, the weight loss was 7.9%, and an endothermic peak was observed at 126.2 ° C (peak). Figure 5-39 As shown, in 1 The presence of acetone was confirmed in the H NMR spectrum, while no n-heptane signal was observed. Based on the results, form L is likely a 2-MeTHF solvate.

[0585] 5.15M

[0586] Form M (3-11-A2) was obtained from liquid vapor diffusion in EtOAc / IPA. Figure 5-40 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-41 As shown in the TGA and DSC data in , up to 150 ° C, the weight loss was 3.7%, and an endothermic peak was observed at 122.6 ° C (peak). Figure 5-42 As shown, in 1 The presence of IPA was confirmed in H NMR, while no EtOAc signal was observed. Based on the results, Form M is likely an IPA solvate.

[0587] 5.16N type

[0588] Form N (3-08-A8) was obtained by slow evaporation in EtOH. Figure 5-43 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-44 As shown in the TGA and DSC data in , up to 150°C, the weight loss was 4.1%, and endothermic peaks were observed at 85.4°C, 126.5°C, and 150.9°C (peak). Figure 5-45 As shown, in 1 The presence of EtOH was confirmed in the H NMR spectrum. Based on the results, Form N is likely an EtOH solvate.

[0589] 5.17O

[0590] Form O (3-08-A11) was obtained by slow evaporation in MIBK. Figure 5-46 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-47 As shown in the TGA and DSC data in , up to 130 ° C, the weight loss was 2.0%, and two endothermic peaks were observed at 106.2 ° C and 151.2 ° C (peak). Figure 5-48 As shown, in1 The presence of MIBK was confirmed in the H NMR spectrum. Based on the results, the O form is likely a MIBK solvate.

[0591] 5.18P

[0592] Form P (3-07-A14) was obtained by slow evaporation in DMF. Figure 5-49 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-50 As shown in the TGA and DSC data in , up to 130 ° C, the weight loss was 8.3%, and an endothermic peak was observed at 89.9 ° C (peak). Figure 5-51 As shown, in 1 The presence of DMF was confirmed in the H NMR spectrum. Based on the results, Form P is likely a DMF solvate.

[0593] 5.19Q type

[0594] Form Q (3-11-A1) was obtained by liquid vapor diffusion in MIBK / n-heptane. Figure 5-52 The XRPD results shown in the figure indicate that the product is crystalline. Figure 5-53 As shown in the TGA and DSC data in , up to 120°C, the weight loss was 6.0%, and endothermic peaks were observed at 92.9°C, 148.9°C, and 170.0°C (peak). Figure 5-54 As shown, in 1 The presence of MIBK was confirmed in the H NMR spectrum, and no n-heptane signal was observed. Based on the results, form Q is likely a MIBK solvate.

[0595] 6. Crystallographic Data and Experiments of Composition 4a

[0596] Experimental. Single colorless plate-like crystals of (composition 4a) were used as received. Suitable crystals (0.28×0.18×0.09) mm were selected. 3 The crystals were kept at T = 173 (2) K during data collection. The structure was solved using Olex2 (Dolomanov et al., 2009) with the XT (Sheldrick, 2015) structure solver using the eigenphase solution method. The model was improved using the XL version (Sheldrick, 2008) using least squares minimization.

[0597] Crystal data. C 65 H 72 Cl2F2N8O 15 ,M r=1314.20,triclinic system,P1(No.1), α=68.1780(10)°, β=69.4150(10)°, γ=87.7760(10)°, ,T=173(2)K,Z=1,Z'=1,μ(MoK α )=0.178, 26758 reflections were measured, 11949 unique reflections (R int =0.0528) for all calculations. The final wR2 was 0.2465 (all data), and R1 was 0.0835 (I>2(I)). p

[0598] Table 6-2: Fractional atomic coordinates of composition 4A (×10 4 ) and equivalent isotropic displacement parameter .U eq Defined as orthogonal U ij 1 / 3 of the trace.

[0599]

[0600]

[0601]

[0602]

[0603] Table 6-3: Anisotropic displacement parameters (×10 4 ) Composition 4A. The anisotropic shift factor index takes the following form: -2π 2 [h 2 a* 2 ×U 11 +...+2hka*×b*×U 12 ]$

[0604]

[0605]

[0606]

[0607] Table 6-3: Bond lengths of composition 4A (in units ).

[0608]

[0609]

[0610]

[0611] Table 6-4: Bond angles of composition 4A.

[0612]

[0613]

[0614]

[0615]

[0616]

[0617] Table 6-5: Hydrogen fraction atomic coordinates of composition 4A (×10 4 ) and the equivalent isotropic displacement parameter ( ). eq Defined as orthogonal U ij 1 / 3 of the trace.

[0618]

[0619]

[0620]

[0621]

[0622] Table 6-4: Hydrogen bonding information of composition 4A.

[0623]

[0624]

[0625] Table 6-5: Atomic occupancies of all atoms not fully occupied in Composition 4A.

[0626]

[0627]

[0628] The foregoing is merely illustrative of the present invention and is not intended to limit the present invention to the disclosed uses. Variations and modifications that are routine to those skilled in the art are intended to fall within the scope and nature of the present invention as defined by the appended claims. All references, patents, applications, and publications mentioned are incorporated herein by reference in their entirety as if written herein.

Claims

1. A composition comprising a compound having formula 4 and a compound having formula B:

2. The composition of claim 1, wherein the compound of formula 4 is a compound of formula 5M:

3. The composition of claim 1, wherein the compound of formula 4 is a compound of formula 5P:

4. The composition of claim 2, wherein the compound of formula B is a compound of formula B1:

5. The composition of claim 2, wherein the compound of formula B is a compound of formula B2:

6. The composition of any one of claims 1-5, wherein the composition comprises the compound of formula 4 and the compound of formula B in a 2:1 ratio.

7. The composition of any one of claims 1-5, wherein the composition further comprises 2-methyltetrahydrofuran.

8. The composition of claim 7, wherein the ratio of 2-methyltetrahydrofuran to the compound of formula B is 2:

1.

9. The composition of claim 1, wherein the composition is:

10. The composition of claim 9, wherein the composition is:

11. The composition of claim 9, wherein the composition is:

12. The composition of any one of claims 9-11, wherein the composition is in a crystalline state.

13. The composition of any one of claims 9-11, wherein the composition is a co-crystal.

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