EA045678B1 - IMPROVED SYNTHESIS OF A KEY INTERMEDIATE TO PRODUCE A G12C KRAS INHIBITOR COMPOUND - Google Patents

Description

Field of technology to which the invention relates

The present invention relates to an improved, efficient, scalable method for preparing intermediates such as compound 5M, characterized by the structure o

’ useful for the synthesis of compounds that suppress G12C KRAS mutations.

Prerequisites for creating the invention

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 patients with NSCLC, and some studies indicate that KRAS mutations are a negative prognostic factor in patients with NSCLC. Recently, mutations in the Kirsten rat sarcoma viral oncogene V-Ki-ras2 homolog (KRAS) homolog were found to confer resistance to epidermal growth factor receptor (EGFR)-targeting therapies in colorectal cancer; accordingly, KRAS mutational status may provide important information before initiating TKI therapy. Taken together, there is a medical need for new ways to treat patients with pancreatic cancer, lung adenocarcinoma, or colorectal cancer, especially for those who have been diagnosed with such cancers characterized by a KRAS mutation, and including those who have progressed after chemotherapy.

Brief description of graphic materials

In fig. 1 shows the crystal configuration of composition 4a.

In fig. Figure 2-1 shows the overlap of X-ray powder patterns for dione racemates types A-E.

In fig. Figure 2-2 shows the overlap of X-ray powder patterns for a (1S)-(-)camphanic acid cocrystal.

In fig. Figures 2-3 show the overlap of X-ray powder diffraction patterns for a cocrystal of (+)-2,3-dibenzoylD-tartaric acid.

In fig. Figures 2-4 show the overlap of X-ray powder patterns for a D-(+)-malic acid cocrystal.

In fig. Figures 2-5 show the overlap of X-ray powder patterns for the M-dione cocrystal at different temperatures.

In fig. Figures 2-6 show the overlap of X-ray powder patterns for a P-dione cocrystal at different temperatures.

In fig. Figures 2-7 show the overlap of X-ray powder patterns for a mixture of an M-dione cocrystal and a P-dione cocrystal at different temperatures.

In fig. Figures 2-8 show the overlap of X-ray powder patterns for the dione racemate at different temperatures (M1).

In fig. Figures 2-9 show the overlap of X-ray powder patterns for the dione racemate at different temperatures (M1).

In fig. Figure 2-10 shows the ternary phase diagram for M/P dione cocrystals.

In fig. Figure 2-11 shows the ternary phase diagram for the M/P dione.

In fig. Figure 3-1 shows the X-ray powder diffraction pattern for the type A dione racemate.

In fig. Figure 3-2 shows the overlap of the TGA/DSC curves for the type A dione racemate.

In fig. Figure 3-3 shows the 1H NMR spectrum for the type A dione racemate.

In fig. Figures 3-4 show a PLM image of the type A dione racemate.

In fig. Figures 3-5 show the X-ray powder diffraction pattern for the type B dione racemate.

In fig. Figures 3-6 show the overlap of the TGA/DSC curves for the type B dione racemate.

In fig. Figures 3-7 show the 1H NMR spectrum for the type B dione racemate.

In fig. Figures 3-8 show the X-ray powder diffraction pattern for the type C dione racemate.

In fig. Figures 3-9 show the overlap of the TGA/DSC curves for the type C dione racemate.

In fig. Figure 3-10 shows the 1H NMR spectrum for the type C dione racemate.

In fig. Figure 3-11 shows the X-ray powder diffraction pattern for the type D dione racemate.

In fig. Figure 3-12 shows the overlap of the TGA/DSC curves for the type D dione racemate.

In fig. Figure 3-13 shows the 1H NMR spectrum for the type D dione racemate.

In fig. Figure 3-14 shows the X-ray powder diffraction pattern for the type E dione racemate.

In fig. Figure 3-15 shows the overlap of the TGA/DSC curves for the type E dione racemate.

In fig. Figure 3-16 shows the 1H NMR spectrum for the type E dione racemate.

In fig. Figure 3-17 shows an X-ray powder diffraction pattern for a type A M-dione cocrystal.

In fig. Figure 3-18 shows the overlap of the TGA/DSC curves for a type A M-dione cocrystal.

In fig. Figure 3-19 shows the 1H NMR spectrum for a type A M-dione cocrystal.

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In fig. Figure 3-20 shows an X-ray powder diffraction pattern for a type A P-dione cocrystal.

In fig. Figure 3-21 shows the overlap of the TGA/DSC curves for a type A P-dione cocrystal.

In fig. Figure 3-22 shows the 1H NMR spectrum for a type A P-dione cocrystal.

In fig. Figure 3-23 shows the overlap of X-ray powder patterns for the dione racemate forms.

In fig. Figures 3-24 show X-ray powder diffraction patterns for competitive suspension samples.

In fig. Figure 3-25 shows the overlap of X-ray powder patterns for the resulting Pdion cocrystal.

In fig. Figure 3-26 shows the overlap of the 1H NMR spectra for the M/P-dione cocrystals.

In fig. Figure 3-27 shows the overlap of X-ray powder patterns for the resulting Pdion cocrystal.

In fig. Figure 4-1 shows the interconversion diagram of the crystal forms of the Mdion cocrystal with DBTA.

In fig. Figure 5-1 shows the overlap of X-ray powder patterns for the crystal forms of the M-dione cocrystal with DBTA (types A-E).

In fig. Figure 5-2 shows the overlap of X-ray powder patterns for the crystal forms of the M-dione cocrystal with DBTA (types F-K).

In fig. Figure 5-3 shows the overlap of X-ray powder patterns for the crystal forms of the M-dione cocrystal with DBTA (types L-Q).

In fig. Figure 5-4 shows a powder X-ray diffraction pattern for Type A.

In fig. Figure 5-5 shows the TGA/DSC curves for Type A.

In fig. Figure 5-6 shows the 1H NMR spectrum for type A.

In fig. Figures 5-7 show the X-ray powder pattern overlap for Type B.

In fig. Figures 5-8 show the TGA/DSC curves for type B.

In fig. 5-9 shows the 1H NMR spectrum for type B.

In fig. Figure 5-10 shows a powder X-ray diffraction pattern for Type C.

In fig. Figures 5-11 show the TGA/DSC curves for Type C.

In fig. Figure 5-12 shows the 1H NMR spectrum for type C.

In fig. Figure 5-13 shows a powder X-ray diffraction pattern for Type D.

In fig. Figures 5-14 show the TGA/DSC curves for Type D.

In fig. Figure 5-15 shows the 1H NMR spectrum for type D.

In fig. Figure 5-16 shows a powder X-ray diffraction pattern for Type E.

In fig. Figure 5-17 shows the TGA/DSC curves for Type E.

In fig. Figure 5-18 shows the 1H NMR spectrum for type E.

In fig. Figure 5-19 shows a powder X-ray diffraction pattern for type F.

In fig. Figure 5-20 shows the TGA/DSC curves for type F.

In fig. Figure 5-21 shows the 1H NMR spectrum for type F.

In fig. Figure 5-22 shows a powder X-ray diffraction pattern for Type G.

In fig. Figure 5-23 shows the TGA/DSC curves for Type G.

In fig. Figure 5-24 shows the 1H NMR spectrum for type G.

In fig. Figure 5-25 shows a powder X-ray diffraction pattern for type H.

In fig. Figures 5-26 show the TGA/DSC curves for type H.

In fig. Figure 5-27 shows the 1H NMR spectrum for type H.

In fig. Figure 5-28 shows a powder X-ray diffraction pattern for Type I.

In fig. Figure 5-29 shows the TGA/DSC curves for Type I.

In fig. Figure 5-30 shows the 1H NMR spectrum for type I.

In fig. Figure 5-31 shows the X-ray powder diffraction pattern for Type J.

In fig. Figure 5-32 shows the TGA/DSC curves for Type J.

In fig. Figure 5-33 shows the 1H NMR spectrum for type J.

In fig. Figure 5-34 shows an X-ray powder diffraction pattern for Type K.

In fig. Figure 5-35 shows the TGA/DSC curves for Type K.

In fig. Figure 5-36 shows the 1H NMR spectrum for type K.

In fig. Figure 5-37 shows a powder X-ray diffraction pattern for Type L.

In fig. Figure 5-38 shows the TGA/DSC curves for type L.

In fig. Figure 5-39 shows the 1H NMR spectrum for type L.

In fig. Figure 5-40 shows a powder X-ray diffraction pattern for type M.

In fig. Figure 5-41 shows the TGA/DSC curves for type M.

In fig. Figure 5-42 shows the 1H NMR spectrum for type M.

In fig. Figure 5-43 shows a powder X-ray diffraction pattern for Type N.

In fig. Figure 5-44 shows the TGA/DSC curves for Type N.

In fig. Figure 5-45 shows the 1H NMR spectrum for type N.

In fig. Figure 5-46 shows an X-ray powder diffraction pattern for type O.

In fig. Figure 5-47 shows the TGA/DSC curves for Type O.

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In fig. Figure 5-48 shows the 1H NMR spectrum for type O.

In fig. Figure 5-49 shows a powder X-ray diffraction pattern for Type P.

In fig. Figure 5-50 shows the TGA/DSC curves for type P.

In fig. Figure 5-51 shows the 1H NMR spectrum for type P.

In fig. Figure 5-52 shows the X-ray powder diffraction pattern for Type Q.

In fig. Figure 5-53 shows the TGA/DSC curves for type Q.

In fig. Figure 5-54 shows the 1H NMR spectrum for type Q.

In fig. 6-1 shows the HPLC results for M-5 when separated using 1,3-diphenyl-3oxopropanesulfonic acid.

In fig. 6-2 shows the HPLC results for 5 (excess P-atropoisomer) when separated using 1,3-diphenyl-3-oxopropanesulfonic acid.

Brief description of the invention

The present invention relates to an improved preparation of a compound having the following chemical structure:

and its key intermediates, i.e. compositions and compounds characterized by the following chemical structures:

The present invention further relates to a method for producing compound 5M, characterized by the following chemical structure:

The present invention further relates to a composition characterized by the structure

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Detailed description

Definitions.

Abbreviations. The following abbreviations may be used in this document

ACN acetonitrile AcOH acetic acid aq. or aq. water VOS or Vos tert-butyloxycarbonyl ViON n-butanol ViOAc butanol acetate ruff cyclopentyl methyl ether SNS1 ₃ trichloromethane DCE 1,2-dichloroethane DABCO 1,4-diazabicyclo[2.2.2]octane DCM dichloromethane DMA N,N-dimethyl acetamide DMAP 4-dimethylaminopyridine DME 1,2-dimethoxyethane DMF N,N-dimethylformamide DMSO dimethyl sulfoxide Dppf, DPPF or dppf 1, G-bis(diphenylphosphino)ferrocene EQ OR EQ. OR EQUIV. equivalent ESI or ES electrospray ionization Et ethyl Et ₂ O diethyl ether EtOAc ethyl acetate EtOH ethanol G gram h. hour H ₂ 0 water HPLC high pressure liquid chromatography iPr isopropyl IPA isopropyl alcohol IP Ac isopropyl acetate iPr ₂ NEt or DIPEA N-ethyldiisopropylamine (Hunig's base) KHMDS potassium hexamethyldisilazide KOAc potassium acetate LDA lithium diisopropylamide 2,4-bis(4-methoxyphenyl)-2,4-dithioxo-1,3,2,4- Lawsson's reagent dithiadiphosphetane, 2,4-bis-(4-methoxyphenyl)-1,3-dithia-2,4- diphosphetane-2,4-disulfide LC MS, LCMS, LC-MS mass spectrometry with liquid chromatography or LC/MS LG leaving group (e.g. halogen, mesylate, triflate) LHMDS or LiHMDS lithium hexamethyldisilazide mass/charge mass divided by charge Me methyl MeCN acetonitrile MeOH methanol Met metal particles intended for reactions cross-couplings (for example, MgX, ZnX, SnR ₃ , SiR ₃ , B(OR) ₂ )

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2-MeTHF 2-methyltetrahydrofuran mg milligram min. minutes MIBK 4-methyl-2-pentanone ml milliliters MS mass spectra MTBE methyl tert-butyl ether n-BuLi n-butyllithium NaHMDS sodium hexamethyldisilazide N.B.S. N-bromosuccinimide NCS N-chlorosuccinimide NMR nuclear magnetic resonance Pd ₂ (dba) ₃ tris(dibenzyl indenacetone)dipalladium(0) Pd(dppf)Cl ₂ DCM [1,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium(P), complex with dichloromethane Pd(PPh ₃ ) ₄ tetrakis(triphenylphosphin)palladium(0) Ph phenyl PR or PG or def. group protective group rbf round bottom flask RP-HPLC reverse phase high pressure liquid chromatography RT or k.t. room temperature us. or sat. saturated SFC supercritical fluid chromatography SPhos Pd G3 or SPhos G3 (2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl) [2-(2'amino-1,1'-biphenyl)] palladium(II) methanesulfonate TBAF tetra-n-butylammonium fluoride TBTU N, A, A', A'-tetramethyl-b?-(benzotriazol-1-yl)uronium tetrafluoroborate ABuOH tert-butanol TEA or Et ₃ N trimethylamine T.F.A. trifluoroacetic acid THF tetrahydrofuran UV XRPD ultraviolet X-ray powder diffractometry

The use of the singular, plural and similar references in the context of the description of the present invention (especially in the context of the claims) should be construed to include both singular and plural forms unless otherwise indicated. The listing of ranges of values in this document is intended only to serve as a shorthand way of referring individually to each individual value falling within a given range unless otherwise noted herein, and each individual value is included in the description as if it were separate. specified in this document. The use of any and all examples or illustrative language (eg, such as) presented herein is intended to better illustrate the present invention and is not intended to limit the scope of the present invention unless otherwise stated. Nothing in the specification should be construed as indicating any unstated element as essential to the practice of the present invention.

As used herein, the term alkyl refers to straight and branched C ₁ -C ₈ hydrocarbon groups including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, t-butyl, n-pentyl , 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl and 2-ethylbutyl . The term C _mn means that the alkyl group contains from m to n carbon atoms. The term alkylene refers to an alkyl group having a substituent. Alkyl (eg methyl) or

- 5 045678 the alkylene (e.g. -CH2-) group may be substituted with one or more and typically one to three substituents independently selected from, for example, halogen, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C ₁ . ₈ αlkulα, C ₂ . ₈ alkenyl, C ₂ . ₈ alkynyl, -NC, amino, -CO2H, -CO ₂ C ₁ -C ₈ -alkyl, -OCOC ₁ -C ₈ -alkyl, C ₃ -C ₁₀ -cycloalkyl, C ₃ -C ₁₀ -heterocycloalkyl, C ₅ - C ₁₀ aryl and C ₅ -C ₁₀ heteroaryl. The term haloalkyl specifically refers to an alkyl group wherein at least one, such as one to six, or all of the hydrogen atoms in the alkyl group are replaced by halogen atoms.

The terms alkenyl and alkynyl indicate an alkyl group that additionally contains a double bond or a triple bond, respectively.

As used herein, the term halogen refers to fluorine, chlorine, bromine and iodine. The term alkoxy is defined as -OR, where R is alkyl.

As used herein, the term amino or amine interchangeably refers to -NR2rpynne, where each R represents, for example, H or a substituent. In some embodiments, the amino group is further substituted to form an ammonium ion, for example, NR ₃ ⁺ . Ammonium moieties are particularly included within the definition of amino or amine. The substituents may be, for example, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, amide or carboxylate. The R group may be further substituted, for example, by one or more, for example one to four, groups selected from halogen, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, carboxylate, amine and amide. An amide or amido group interchangeably refers to a group similar to an amine or amino group but additionally containing C(O), for example -C(O)NR2.

As used herein, the term aryl refers to C ₆ . ₁₄ monocyclic or polycyclic aromatic group, preferably C ₆ . ₁₀ monocyclic or bicyclic aromatic group or to C ₁₀ . ₁ 4polycyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Aryl is also classified as _C10 . ₁₄ bicyclic and tricyclic carbon rings, where one ring is aromatic and the others are saturated, partially unsaturated or aromatic, for example dihydronaphthyl, indenyl, indanyl or tetrahydronaphthyl (tetralinyl). Unless otherwise specified, the aryl group may be unsubstituted or may be substituted by one or more, and in particular one to four, groups independently selected from, for example, halogen, C ₁ . ₈ alkyl, C ₂ . ₈ αlkenyl, C ₂ . ₈ alkynyl, -CF ₃ , -OCF ₃ , -NO ₂ , -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO ₂ C ₁ -C ₈ -alkyl, -OCOC ₁ -C ₈ -alkyl , C ₃ -C ₁₀ -cycloalkyl, C ₃ -C ₁₀ -heterocycloalkyl, C ₅ -C ₁₀ -aryl and C ₅ -C ₁₀ -heteroaryl.

As used herein, the term cycloalkyl refers to a monocyclic or polycyclic non-aromatic carbocyclic ring, where the polycyclic ring may be a fused, bridged ring or a spiro ring. A carbocyclic ring can contain from 3 to 10 carbon ring atoms. Provided carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and cyclononyl.

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) atoms in total, of which one to five (eg, 1, 2, 3, 4 or 5) atoms are independently selected from nitrogen, oxygen and sulfur. Non-limiting examples of heterocycloalkyl groups include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, dihydropyrrolyl, morpholinyl, thiomorpholinyl, dihydropyridinyl, oxacycloheptyl, dioxacycloheptyl, thiacycloheptyl, and diazacycloheptyl.

Unless otherwise specified, a cycloalkyl or heterocycloalkyl group may be unsubstituted or may be substituted by one or more, and in particular by one to four groups. Some contemplated substituents include halogen, C ₁ . ₈ αlkyl, C ₂ . ₈ alkenyl, C ₂ . ₈ alkynyl, -OCF ₃ , -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO ₂ C ₁ -C ₈ -alkyl, -OCOC ₁ -C ₈ -alkyl, C ₃ -C ₁₀ -cycloalkyl, C ₃ C ₁₀ -heterocycloalkyl, C ₅ -C ₁₀ -aryl and C ₅ -C ₁₀ -heteroaryl.

As used herein, the term heteroaryl refers to a monocyclic or polycyclic ring system (e.g., bicyclic) containing one to three aromatic rings and containing one to four (e.g., 1, 2, 3, or 4) heteroatoms selected from nitrogen, oxygen and sulfur in the aromatic ring. In some embodiments, the heteroaryl group contains from 5 to 20, from 5 to 15, from 5 to 10, or from 5 to 7 ring atoms. Heteroaryl also refers to _C10 . ₁₄ bicyclic and tricyclic rings, where one ring is aromatic and the others are saturated, partially unsaturated or aromatic. Examples of heteroaryl groups 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, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzothiophenyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, o xazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidyl and thienopyridyl. Unless otherwise specified, the heteroaryl group may be unsubstituted or may be substituted by one or more, and in particular one to four or one or two substituents. Substituents contemplated include halogen, C _1-8 alkyl, C _2-8 αlkenyl, C^alkynyl, -OCF ₃ , -NO2, -CN, -NC, -OH, alkoxy, amino, -CO2H, -CO ₂ C ₁ - C ₈ -alkyl, -OCOC ₁ -C ₈ -alkyl, C ₃ -C ₁₀ -cycloalkyl, C ₃ -C ₁₀ -heterocycloalkyl, C ₅ -C ₁₀ -aryl and C ₅ -C ₁₀ -heteroaryl.

As used in this document, the term Boc refers to the structure

Embodiments

Option 1.

In one embodiment of the present invention, the present invention provides a composition, wherein the composition contains a compound of formula 4,

and a compound of formula B,

Embodiment 2.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 1, wherein the compound of Formula 4 is a compound of Formula 5M:

Embodiment 3.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 1, wherein the compound of Formula 4 is a compound of Formula 5P:

Embodiment 4.

In another embodiment of the present invention, the present invention provides a composition according to any of embodiments 1-3, wherein the compound of formula B is a compound of formula B1:

Ph but ₂ s with ₂ n. R

Phbi.

Embodiment 5.

In another embodiment of the present invention, the present invention provides a composition according to any of embodiments 1-3, wherein the compound of formula B is a compound of formula B2:

Ph

Vo CO ₂ H )—ί R

BUT ₂ C

Ph B2.

Embodiment 6.

In another embodiment of the present invention, the present invention provides a composition according to any of Embodiments 1-5, wherein the composition comprises a 2 to 1 ratio of a compound of Formula 4 to a compound of Formula B.

Embodiment 7.

In another embodiment of the present invention, the present invention provides a composition according to any of embodiments 1-6, wherein the composition further comprises 2-methyltetrahydrofuran having the formula

Embodiment 8.

In another embodiment, the present invention provides a composition according to any of embodiments 1-7, wherein the ratio of 2methyltetrahydrofuran to the compound of formula B is 2 to 1.

Embodiment 9.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 1, wherein the composition is characterized by the formula

Embodiment 10.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 9, wherein the composition is characterized by the formula

Embodiment 11.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 9, wherein the composition is characterized by the formula

Embodiment 12.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 9, wherein the composition is characterized by the formula

Embodiment 13.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 9, wherein the composition is characterized by the formula

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Embodiment 14.

In another embodiment of the present invention, the present invention provides a composition according to any of embodiments 1-13, wherein the composition is in a crystalline state.

Embodiment 15.

In another embodiment of the present invention, the present invention provides a method for preparing a composition of formula 4a, the method comprising reacting compound 4 having the following chemical structure:

with compound B1 characterized by the formula

Ph

Ph in the presence of 2-methyltetrahydrofuran to form a composition of formula 4a characterized by the structure

Embodiment 16.

In another embodiment of the present invention, the present invention provides a method for preparing a compound of Formula 5M having the following chemical structure:

the method includes:

a) carrying out the reaction of compound 4, characterized by the following chemical structure:

Ph co ₂ n ί P no _g s

4, with compound B1 characterized by the formula Ph in the presence of 2 methyltetrahydrofuran to form

compositions of formula 4a characterized by the structure

4a „ ’ presented in the form of crystals;

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b) isolating composition 4a and

c) treating the isolated composition 4a with a base to obtain the compound of formula 5M.

Embodiment 17.

In another embodiment of the present invention, the present invention provides a method according to Embodiment 16, wherein the base is Na ₂ HPO ₄ .

Embodiment 18.

In another embodiment of the present invention, the present invention provides a method according to Embodiment 16, wherein the base is NaHCO3.

Embodiment 19.

In another embodiment of the present invention, the present invention provides a composition, wherein the composition contains a compound of formula 4,

and a compound of formula 11,

Embodiment 20.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 19, wherein the compound of Formula 4 is a compound of Formula 5M:

Embodiment 21.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 19, wherein the compound of Formula 4 is a compound of Formula 5P:

Embodiment 22.

In another embodiment of the present invention, the present invention provides a composition according to any of embodiments 19-21, wherein the compound of formula 11 is a compound of formula 11a:

Embodiment 23.

In another embodiment of the present invention, the present invention provides a composition according to any of embodiments 19-21, wherein the compound of formula 11 is a compound of formula 11b:

Embodiment 24.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 19, wherein the composition is characterized by the formula

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Embodiment 25.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 19, wherein the composition is characterized by the formula

Embodiment 26.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 19, wherein the composition is characterized by the formula

Embodiment 27.

In another embodiment of the present invention, the present invention provides a composition according to Embodiment 19, wherein the composition is characterized by the formula

Embodiment 28.

In another embodiment, the present invention provides a composition according to any of embodiments 19-27, wherein the composition comprises a 1 to 1 ratio of a compound of Formula 4 to a compound of Formula 11.

Embodiment 29.

In another embodiment of the present invention, the present invention provides a method according to Embodiment 16, wherein a compound of Formula 5M is used to prepare a compound of Formula 9:

Embodiment 30.

The method of Embodiment 29, wherein the method further comprises mixing a compound of Formula 9 with at least one pharmaceutically acceptable excipient to form a pharmaceutical composition.

Connections in accordance with the present invention

Provided herein are KRAS inhibitors characterized by the structures discussed in more detail below.

The compounds disclosed herein include all pharmaceutically acceptable isotopically labeled compounds wherein one or more atoms of the compounds disclosed herein are replaced by atoms having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number numbers commonly found in nature. Examples of isotopes that may be included in the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, such as ² H, ³ H, 11 C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N , ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²³ I and ¹²⁵ I, respectively. Such radiolabeled compounds may

- 11 045678 be used to help determine or measure the effectiveness of compounds by describing, for example, the site or mechanism of action or binding affinity to a pharmacologically important site of action. Certain isotope-labeled compounds of the present invention, for example, compounds into which a radioactive isotope is included, are useful in drug and/or substrate tissue distribution studies. Radioactive isotopes tritium, i.e. ³ H, and carbon-14, i.e. ¹⁴ C are particularly suitable for this purpose given their ease of administration and ready means for their detection.

Substitution with heavier isotopes such as deuterium, i.e. 2H may provide some therapeutic benefits resulting from greater resistance to inactivation by metabolism, such as increased in vivo half-life or reduced dosage requirements, and is therefore preferred in some circumstances.

Substitution with positron-active isotopes such as 11C, ^18F , ^15O and ^13N can be used in positron emission topography (PET) studies to determine the extent of receptor occupancy by a substrate. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art, or by methods similar to those described in the Preparations and Examples sections, as set forth below, using the appropriate labeled isotope of the reagent instead of the unlabeled reagent used previously.

The isotopically labeled compounds disclosed herein generally can be prepared by conventional techniques known to those skilled in the art, or by methods similar to those described in the accompanying Examples and Schemes using the appropriate isotopically labeled reagent in place of the unlabeled reagent. reagent used previously.

Some of the compounds disclosed herein may exist as stereoisomers (ie, isomers that differ only in the spatial arrangement of the atoms), including optical isomers and conformational isomers (or conformers). The compounds disclosed herein include all stereoisomers, both as pure preparations of the individual stereoisomers and as fortified preparations of each stereoisomer, and both racemic mixtures of such stereoisomers and the individual diastereomers and enantiomers, which can be separated according to methods which are known to those skilled in the art. In addition, the compounds disclosed herein include all tautomeric forms of the compounds.

Some of the compounds disclosed herein may exist as atropoisomers, which are conformational stereoisomers that occur when rotation around a single bond in a molecule is prevented or greatly slowed down by steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers, both as pure preparations of the individual atropisomers, fortified preparations of each atropisomer, or a nonspecific mixture of each atropisomer. In cases where the rotational barrier around a single bond is sufficiently high and the interconversion between conformations is sufficiently slow, then separation and isolation of isomeric species of molecules may be allowed. For example, groups such as, but not limited to, the following groups:

The term monohydrate means a salt of compound 9 having approximately one associated water molecule. Those skilled in the art will appreciate that the exact number of bound water molecules may vary slightly at any given time with changing temperature, pressure and other environmental influences. All minor changes in the number of bound water molecules are intended to be within the scope of the present invention.

The term dihydrate means a salt of compound 9 having approximately two associated water molecules. Those skilled in the art will appreciate that the exact number of bound water molecules may vary slightly at any given time with changing temperature, pressure and other environmental influences. All minor changes in the number of bound water molecules are intended to be within the scope of the present invention.

The term cocrystal means a crystalline material containing two or more compounds at ambient temperature (20 to 25°C, preferably 20°C), of which at least two are held together by weak interaction, where at least one of the compounds is cocrystal former, and the other is compound 5. A weak interaction is defined as an interaction that is neither ionic nor covalent and includes, for example, hydrogen bonds, van der Waals forces, and π-π interactions.

The term amorphous form or amorphous means a material that lacks long-range order and therefore does not exhibit distinct X-ray diffraction peaks, that is, a Bragg diffraction peak. The X-ray powder diffraction pattern of an amorphous material is characterized by one or more amorphous halos.

The term amorphous halo is a roughly bell-shaped maximum in the X-ray powder diffraction pattern of an amorphous substance.

The term substantially pure refers to a solid form of compound 9 having a purity of greater than about 95%, in particular greater than about 99.5%, more particularly greater than about 99.8%, and even more particularly greater than about 99.9%.

The term patient refers to animals such as dogs, cats, cows, horses, sheep and humans. Specific patients are mammals. The term patient includes males and females.

The terms treating, treating, or administering treatment, etc. include preventive (eg, prophylactic) and palliative treatment.

The term excipient means any pharmaceutically acceptable additive, carrier, diluent, excipient or other ingredient, other than an active pharmaceutical ingredient (API), that is typically included for formulation and/or administration to a patient.

Pharmaceutical compositions, dosage and routes of administration

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

Suitable pharmaceutical formulations can be determined by one skilled in the art depending on the route of administration and the required dose. See, for example, Remington's Pharmaceutical Sciences, 1435-712 (18th ed., Mack Publishing Co, Easton, PA, 1990). The formulations may affect the physical state, stability, in vivo release rate, and in vivo elimination rate of the administered agents. Depending on the route of administration, the appropriate dose may be calculated according to body weight, body surface area or organ size. It is customary for those skilled in the art to further refine the calculations necessary to determine the appropriate treatment dose without undue experimentation, especially in light of the dosage information and assays disclosed herein, as well as the pharmacokinetic data that can be obtained from clinical trials in animals or people.

The phrases pharmaceutically acceptable or pharmacologically acceptable refer to molecular substances and compositions that do not cause unwanted, allergic or other adverse reactions when administered to an animal or human. As used herein, the term pharmaceutically acceptable includes all possible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption retarding agents, etc. The use of such excipients for pharmaceutically active substances is widely known in the art. Unless any conventional media or agent is incompatible with the therapeutic compositions, their use in the therapeutic compositions is intended. Additional active ingredients may also be included in the compositions. In illustrative embodiments, the formulation may contain corn syrup powder, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tribasic calcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine , L-valine, dibasic potassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine , ascorbic acid, calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate , sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, copper(II) sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium trichloride, sodium molybdate, vitamin D3 and cyanocobalamin.

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

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

Pharmaceutically acceptable acid addition salts include salts of inorganic or organic acids. Examples of suitable acid addition 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, those formed with formic, acetic, citric, oxalic, tartaric or mandelic acids, with hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic carboxylic, sulfonic, sulfo- or phosphoric acids or N-substituted sulfamic acids, for example acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid , tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid acid, nicotinic acid or isonicotinic acid and with amino acids such as the 20 alpha amino acids involved in the synthesis of proteins in nature, for example glutamic acid or aspartic acid, as well as with phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 2- or 3-phosphoglycerate, glucose-6-phosphate, Ncyclohexylsulfamic acid (to form cyclamates) or with other acidic organic compounds such as ascorbic acid.

Pharmaceutical compositions containing the compounds disclosed herein can be prepared in conventional manner, for example, through conventional mixing, dissolving, granulating, dragee-making, trituration, emulsification, encapsulation, entrapment, or lyophilization processes. The appropriate formulation depends on the route of administration chosen.

Suitable compositions for oral administration can be readily formulated by combining the compound disclosed herein with pharmaceutically acceptable excipients, such as carriers well known in the art. Such excipients and carriers allow the compounds of the present invention to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral administration by the patient to be treated. Pharmaceutical preparations for oral use can be prepared by adding a solid excipient to a compound disclosed herein, optionally grinding the resulting mixture, and processing the mixture into granules after adding suitable excipients, if necessary, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulosic preparations. If necessary, substances can be added to improve the disintegration of tablets. Pharmaceutically acceptable ingredients are well known for various types of formulations and may include, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coating materials, preservatives, colorants, thickeners, excipients, antimicrobials, antioxidants and carriers for various types of formulations.

If a therapeutically effective amount of a compound disclosed herein is administered orally, the composition will typically be in the form of a solid formulation (e.g., tablet, capsule, pellet, powder or lozenge) or liquid formulation (e.g., aqueous suspension, solution, tincture or syrup).

When administered in tablet form, the composition may further comprise a functional solid and/or a solid carrier such as gelatin or an excipient. The tablet, capsule and powder may contain from about 1 to about 95% of the compound, and preferably from about 15 to about 90% of the compound.

When administered in the form of a liquid or suspension, a functional liquid and/or a liquid carrier such as water, a hydrocarbon carrier, or oils of animal or plant origin may be added. The liquid form of the composition may further contain saline, solutions of sugar alcohols, solutions of dextrose or other saccharides or glycols. When administered in liquid or suspension form, the composition may contain from about 0.5 to about 90% by weight of a compound disclosed herein, and preferably from about 1 to about 50% by weight of a compound disclosed herein. In one embodiment, the liquid carrier is provided to be non-aqueous or substantially non-aqueous. The composition, intended for administration in liquid form, may be supplied as a rapidly dissolving solid formulation or as a suspension for dissolution immediately prior to administration.

If a therapeutically effective amount of a compound disclosed herein is administered

- 14 045678 by intravenous, dermal or subcutaneous injection, the composition is in the form of a pyrogen-free aqueous solution suitable for parenteral administration. The preparation of such parenterally acceptable solutions having appropriate pH, isotonicity, stability, etc. is within the skill of one skilled in the art. A preferred composition for intravenous, dermal, or subcutaneous injection typically contains, in addition to the compound disclosed herein, an isotonic carrier medium. Such compositions can be prepared for administration as solutions of the free base or pharmacologically acceptable salts in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be obtained in glycerin, liquid polyethylene glycols and mixtures thereof, as well as in oils. Under normal conditions of storage and use, such preparations may not necessarily contain a preservative to prevent the growth of microorganisms.

Injectable compositions may include sterile aqueous solutions, suspensions or dispersions and sterile powders for the immediate preparation of sterile injectable solutions, suspensions or dispersions. In all embodiments, the form must be sterile and must be fluid to the extent that it can be easily syringed. It must be stable under the conditions of manufacture and storage and must resist the contaminating action of microorganisms such as bacteria and fungi through the optional inclusion of a preservative. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (eg, glycerin, propylene glycol and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. In one embodiment, the carrier is provided to be non-aqueous or substantially non-aqueous. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by maintaining the desired particle size of the compound in the dispersion embodiment, and by the use of surfactants. Prevention of exposure to microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thiomersal, etc. In many embodiments, it will be preferred to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of injectable compositions can be achieved by using absorption retarding agents in the compositions, for example aluminum monostearate and gelatin.

Sterile injection solutions are prepared by supplementing the active compounds in the required quantity in an appropriate diluent as needed with the various other ingredients listed above, followed by sterilization by filtration. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile carrier medium that contains the basic dispersion medium and the required other ingredients listed above. In the embodiment of sterile powders intended for the preparation of sterile injectable solutions, preferred production methods are vacuum drying and freeze drying techniques that provide a powder of the active ingredient plus any additional required ingredient from a pre-sterile filtered solution thereof.

Slow-release or sustained-release formulations may also be prepared to achieve controlled release of the active compound in contact with body fluids in the gastrointestinal tract and to provide a substantially constant and effective plasma level of the active compound. For example, release may be controlled by one or more of dissolution, diffusion, and ion exchange. Additionally, a slow-release approach may enhance absorption through saturable or limiting pathways in the gastrointestinal tract. For example, for this purpose, the compound may be incorporated into a polymer matrix of a biodegradable polymer, a water-soluble polymer, or a mixture of both and optionally suitable surfactants. In this context, embedding may mean the incorporation of microparticles into a matrix of polymers. Controlled release formulations are also prepared by encapsulating dispersed microparticles or emulsified microdroplets using known dispersion or emulsion coating technologies.

For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of a nebulized aerosol from pressurized packs or by nebulizer using a suitable propellant. In a pressurized aerosol embodiment, a dosage unit may be defined by providing a valve to deliver a metered amount. For use in an inhaler or insufflator, capsules and cartridges, for example gelatin, can be formulated containing a powder mixture of the compound and a suitable powder base such as lactose or starch.

The compounds disclosed herein may be formulated for parenteral administration by injection (eg, bolus injection or continuous infusion). Injectable formulations may be presented in unit dosage form (eg, ampoules or multi-dose containers) with an added preservative. The compositions may take forms such as suspensions, solutions or emulsions in oily or aqueous carrier vehicles, and may contain formulation aids such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical compositions intended for parenteral administration include aqueous solutions of the compounds in water-soluble form. In addition, suspensions of the compounds can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or carrier media include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compounds and provide highly concentrated solutions. Alternatively, the composition of the present invention may be in the form of a powder for dilution with a suitable carrier medium (eg, sterile pyrogen-free water) before use.

The compounds disclosed herein may also be formulated into compositions for rectal administration, such as suppositories or retention enemas (eg, containing traditional suppository bases). In addition to the formulations described previously, the compounds may also be formulated as a depot formulation. Such long-acting formulations can be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated using suitable polymeric or hydrophobic materials (eg, as an emulsion in a suitable oil) or ion exchange resins, or as sparingly soluble derivatives, eg, as a sparingly soluble salt.

In particular, the compound disclosed herein can be administered orally, buccally or sublingually in the form of tablets containing excipients such as starch or lactose, or in capsules or vaginal suppositories, either alone or mixed with excipients, or in in the form of tinctures or suspensions containing flavoring or coloring agents. Such liquid preparations can be prepared using pharmaceutically acceptable additives such as suspending agents. The compound can also be administered parenterally, for example intravenously, intramuscularly, subcutaneously or intracoronarily. For parenteral administration, the compound is best administered in the form of a sterile aqueous solution, which may contain other substances, such as salts or sugar alcohols such as mannitol or glucose, to render the solution isotonic with blood.

For veterinary use, the compound disclosed herein is administered in a suitable acceptable formulation in accordance with normal veterinary practice. The veterinarian can easily determine the dosage regimen and route of administration that is most appropriate for a particular animal.

In some embodiments, all of the necessary components for treating a KRAS disorder using a compound disclosed herein, either alone or in combination with another agent, or for an intervention traditionally performed to treat such disorder, may be packaged in a kit . Specifically, the present invention provides a kit for use in a therapeutic intervention for a disease, containing a packaged set of drugs that include a compound disclosed herein, as well as buffers and other components intended to obtain the delivered forms of these drugs, and/or devices for the delivery of such drugs, and/or any agents that are used in combination therapy with a compound disclosed herein and/or instructions for the treatment of a disease contained in the packaging of the drugs. The instructions may be recorded on any tangible medium, such as printed paper or computer-readable magnetic or optical media, or the instructions may be provided in the form of a link to a remote computer data source, such as a World Wide Web page accessible via the Internet.

A therapeutically effective amount means an amount effective to treat or prevent the development of or reduce the severity of existing symptoms in the subject being treated. Determination of effective amounts is within the skill of those skilled in the art, especially in light of the detailed disclosure presented herein. Generally, a therapeutically effective dose refers to that amount of a compound that produces the desired effect. For example, in one preferred embodiment, a therapeutically effective amount of a compound disclosed herein reduces KRAS activity by at least 5% compared to control, by at least 10%, by at least 15%, by 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 at least 65%, at least 70%, at least 75%, at least 80%, at least 85% or at least 90%.

The amount of compound administered may depend on the subject being treated,

- 16 045678 age, health status, sex and weight of the subject, type of simultaneous treatment (if used), severity of the disease, nature of the desired effect, method and frequency of treatment and the decision of the attending physician. Dosing frequency may also depend on pharmacodynamic effects on arterial oxygen pressure. However, the most preferred dosage may be tailored to the individual subject as will be understood and can be determined by one skilled in the art without undue experimentation. This usually involves adjusting the standard dose (eg, reducing the dose if the patient has low body weight).

Although individual needs vary, determination of optimal ranges of effective amounts of a compound is within the skill of those skilled in the art. For administration to humans for the definitive or prophylactic treatment of conditions and disorders identified herein, for example, typical doses of the compounds of the present invention may range from about 0.05 mg/kg/day to about 50 mg/kg/day, such as at least 0.05 mg/kg, at least 0.08 mg/kg, at least 0.1 mg/kg, at least 0.2 mg/kg, at least 0.3 mg/kg, at least 0.4 mg/kg or at least 0.5 mg/kg and preferably 50 mg/kg or less, 40 mg/kg or less, 30 mg/kg or less, 20 mg/kg or less or 10 mg/kg or less, which may be, for example, 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, dosages of the compounds may range from about 0.1 mg/kg/day to about 50 mg/kg/day, from about 0.05 mg/kg/day to about 10 mg/kg/day, from about 0.05 mg /kg/day to about 5 mg/kg/day, from about 0.05 mg/kg/day to about 3 mg/kg/day, from about 0.07 mg/kg/day to about 3 mg/kg/day , from about 0.09 mg/kg/day to about 3 mg/kg/day, from about 0.05 mg/kg/day to about 0.1 mg/kg/day, from about 0.1 mg/kg/day day to about 1 mg/kg/day, from about 1 mg/kg/day to about 10 mg/kg/day, from about 1 mg/kg/day to about 5 mg/kg/day, from about 1 mg/kg /day to about 3 mg/kg/day, from about 3 mg/day to about 500 mg/day, from about 5 mg/day to about 250 mg/day, from about 10 mg/day to about 100 mg/day, from about 3 mg/day to about 10 mg/day, or from about 100 mg/day to about 250 mg/day. Such doses may be administered in a single dose or they may be divided into several doses.

Methods of using G12C KRAS inhibitors.

The present invention provides a method of inhibiting RAS-mediated cell signaling, comprising contacting the cell with an effective amount of one or more compounds disclosed herein. Suppression of RAS-mediated signaling can be assessed and demonstrated by a wide range of pathways known in the art. Non-limiting examples include demonstrating (a) decreased GTPase activity of RAS; (b) decreasing the binding affinity of GTP or increasing the binding affinity of GDP; (c) increasing Koff GTP or decreasing Koff GDP; (d) reducing the levels of signal transduction molecules located downstream in the RAS pathway, for example, reducing the levels of pMEK, pERK or pAKT; and/or (e) reducing the extent of binding of the RAS complex to downstream signaling molecules, including but not limited to Raf. Kits and commercially available assays can be used to determine one or more of the above.

The disclosure also provides methods of using the compounds or pharmaceutical compositions of the present invention to treat disease states, including, without limitation, conditions associated with the G12C KRAS, HRAS, or NRAS mutation (eg, cancer).

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

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

The disclosed compounds inhibit anchorage protein-independent cell growth and therefore have the potential to inhibit tumor metastasis. Accordingly, in another embodiment, the present invention provides a method of inhibiting tumor metastasis, the method comprising administering an effective amount of a compound disclosed herein.

- 17 045678 mente.

G12C KRAS, HRAS, or NRAS mutations have also been identified in hematologic malignancies (eg, cancers that involve the blood, bone marrow, and/or lymph nodes). Accordingly, some embodiments are directed to administering the disclosed compounds (eg, in the form of a pharmaceutical composition) to a patient in need of treatment for hematological malignancies. Such cancers include, but are not limited to, leukemias and lymphomas. For example, the compounds disclosed herein can be used to treat diseases such as acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocyte lymphoma (SLL), chronic myelogenous leukemia (CML), acute monocytic leukemia (AMoL) and/or other types of leukemia. In other embodiments, the compounds are useful for treating lymphomas, such as all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphomas. In various embodiments, the compounds are useful for treating plasma cell malignancies such as multiple myeloma, mantle cell lymphoma, and Waldenström's macroglobulinemia.

Determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation can be made by assessing the nucleotide sequence encoding a KRAS, HRAS, or NRAS protein, by assessing the amino acid sequence of a KRAS, HRAS, or NRAS protein, or by assessing the characteristics of a putative mutant KRAS protein , HRAS or NRAS. The sequence of wild type human KRAS, HRAS or NRAS is known in the art (eg accession no. NP203524).

Methods for detecting a mutation in a KRAS, HRAS, or NRAS nucleotide sequence are known to those skilled in the art. Such methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) assays, polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) assays, real-time PCR assays, sequencing of PCR products, assays using mutant allele specific PCR amplification (MASA), direct sequencing, primer extension reactions, electrophoresis, oligonucleotide probe ligation, hybridization assays, TaqMan assays, SNP genotyping, high resolution melting assays ability and microarray analyses. In some embodiments, samples are assessed for G12C KRAS, HRAS, or NRAS mutations using real-time PCR. Real-time PCR uses fluorescent probes specific for the G12C KRAS, HRAS, or NRAS mutations. When a mutation is present, the probe binds and fluorescence is detected. In some embodiments, the G12C mutation of KRAS, HRAS, or NRAS is identified using a method of direct sequencing of specific regions (eg, exon 2 and/or exon 3) in the KRAS, HRAS, or NRAS gene. This technique will allow us to identify all possible mutations in the sequenced region.

Methods for detecting mutations in a KRAS, HRAS, or NRAS protein are known to those skilled in the art. Such methods include, but are not limited to, detection of mutant KRAS, HRAS, or NRAS using a binding agent (eg, antibody) specific for the mutant protein, protein electrophoresis and Western blotting, and direct peptide sequencing.

Methods for determining whether a tumor or cancer contains a G12C KRAS, HRAS, or NRAS mutation may use a number of samples. In some embodiments, the sample is collected from a subject having a tumor or cancer. In some embodiments, the sample is a fresh tumor/cancer sample. In some embodiments, the sample is a frozen tumor/cancer sample. In some embodiments, the sample is a formalin-fixed, paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

The present invention also provides a method of treating a hyperproliferative disorder in a mammal, which comprises administering to said mammal a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof. In some embodiments, the method relates to treating a subject who is suffering from cancer, such as acute myeloid leukemia, adolescent cancer, childhood adrenal cancer, AIDS-related cancers (eg, lymphoma and Kaposi's sarcoma), anal cancer , appendix cancer, astrocytomas, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumors, Burkitt's lymphoma, carcinoid tumor, atypical teratoid , embryonal tumors, embryonal cell tumor, primary lymphoma, cervical cancer, childhood cancers, chordoma, cardiac tumors, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative disorders, cancer

- 18 045678 colon, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, Ewing's sarcoma, extracranial embryonal cell tumor , extragonadal germinal cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastrointestinal cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumors (GIST), germinal cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer , heart cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet tumors, pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), cancer lung, lymphoma, metastatic squamous cell carcinoma of the neck with an occult primary disease, midline carcinoma, oral cancer, multiple endocrine neoplasia syndromes, multiple myeloma/plasma cell neoplasia, mycosis fungoides, myelodysplastic syndromes, myelodysplastic/myeloproliferative neoplasias, multiple myeloma, Merkel cell carcinoma , malignant mesothelioma, malignant fibrous histiocytoma of bone and osteosarcoma, nasal and sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, cancer pancreas, papillomatosis, paraganglioma, cancer of the paranasal sinuses and nasal cavity, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, gastric cancer (GIT), small cell lung cancer, small intestinal cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymus cancer, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, uncommon childhood cancers, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer. In some embodiments, the method relates to the treatment of a non-cancerous hyperproliferative disorder, such as benign cutaneous hyperplasia (eg, psoriasis), restenosis, or a prostate-related disorder (eg, benign prostatic hypertrophy (BPH)).

In some embodiments, the treatment methods are directed to the treatment of lung cancers, the methods comprising administering an effective amount of any of the above-described compounds (or a pharmaceutical composition containing the same) to a subject in need thereof. In some 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 types of lung cancer that can be treated with the disclosed compounds include, but are not limited to, glandular tumors, carcinoid tumors, and undifferentiated cancers.

The present invention further provides methods for modulating the activity of a G12C mutant KRAS, HRAS, or NRAS protein by contacting the protein with an effective amount of a compound of the present invention. Modulation can be the suppression or activation of protein activity. In some embodiments, the present invention provides methods for inhibiting protein activity by contacting a G12C mutant KRAS, HRAS, or NRAS protein with an effective amount of a compound of the present invention in solution. In some embodiments, the present invention provides methods for inhibiting the activity of a G12C mutant KRAS, HRAS, or NRAS protein by bringing a cell, tissue, or organ that expresses the protein of interest into contact. In some embodiments, the present invention provides methods for inhibiting protein activity in a subject, including, but not limited to, rodents and mammals (eg, humans), by administering to the subject an effective amount of a compound of the present invention. In some embodiments, the modulation percentage is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%. In some embodiments, the suppression percentage is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in a cell by contacting said cell with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in the cell. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in tissue by contacting said tissue with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in said tissue. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in an organism by contacting said organism with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in the organism. In some variants

- 19 045678 In each embodiment, the present invention provides methods for inhibiting G12C KRAS, HRAS or NRAS activity in an animal by contacting said animal with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS or NRAS activity in said animal. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in a mammal by contacting said mammal with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in said mammal. In some embodiments, the present invention provides methods for inhibiting G12C KRAS, HRAS, or NRAS activity in a human by contacting the individual with an amount of a compound of the present invention sufficient to inhibit G12C KRAS, HRAS, or NRAS activity in the individual. The present invention provides methods for treating a disease mediated by G12C KRAS, HRAS or NRAS activity in a subject in need of such treatment.

Combination therapy.

The present invention also provides methods for implementing combination therapy options in which an agent known to modulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes, is used in combination with a compound of the present invention or a pharmaceutically acceptable formulation thereof. salt. In one aspect, such therapy includes, without limitation, the combination of one or more compounds of the present invention with chemotherapeutic agents, therapeutic antibodies, and radiation therapy designed to provide a synergistic or additive therapeutic effect.

A variety of chemotherapeutic agents are currently known in the art and can be used in combination with the compounds of the present invention. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples include chemotherapeutic agents, cytotoxic agents and non-peptide small molecules such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), Venclexta™ (venetoclax) and Adriamycin™ (doxorubicin), as well as a variety of chemotherapy agents. Non-limiting examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide (Cytoxan™); alkylsulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa and uredopa; ethylene imines and methyl melamines, including altretamine, triethylene melamine, triethylene phosphoramide, triethylene thiophosphoramide and trimethylol melamine; nitrogen mustards, such as chlorambucil, chlornaphazine, chlorocyclophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembiquin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas such as carmustine, chlorzotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomycins, actinomycin, anthramycin, azaserine, bleomycins, cactinomycin, calicheycin, carabicin, carminomycin, carcinophilin, Casodex™, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin , esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, cinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprin, thioguanine; pyrimidine analogues such as ancytabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocytabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepithiostane, testolactone; inhibitors of adrenal hormone synthesis, such as aminoglutethimide, mitotane, trilostane; folic acid compensators such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquon; elfomitin; elliptinium acetate; ethoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamole; nitracrine; pentostatin; phenomet; pirarubicin; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK; razoxane; sisofiran; spirogermanium; tenuazonic acid; triaziquon; 2,2',2-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustin; mitobronitol; mitolactol; pipobromance; gacytosine; arabinoside (Ara-C); cyclophosphamide; thiotepa; taxanes, such as paclitaxel and docetaxel; retinoic acid; esperamycins; capecitabine and pharmaceutically acceptable salts, acids or derivatives of any of the above.

Also as suitable chemotherapeutic substances that improve cell health,

- 20 045678 includes antihormonal agents that act to regulate or inhibit the action of hormones on tumors, such as antiestrogens, including, for example, tamoxifen, (Nolvadex™), raloxifene, aromatase inhibitory 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone and toremifene (Fareston); as well as antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogues 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).

If necessary, the compounds or pharmaceutical composition of the present invention can be used in combination with commonly prescribed anti-cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, Avicin, Abagovomab, Acridine carboxamide, adecatumumab, 17-K-allylamino-17-demethoxygeldanamycin, alpharadine, alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, CD22 immunotoxins, antitumor, antitumorigenic herbs, apaziquone, atiprimod, azathioprine, belotecan , bendamustine, BIBW 2992, biricodar, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), calyculin, non-specific cell cycle antitumor agents, dichloroacetic acid, discodermolide, elsamitrucin, enocytabine, epothilone, eribulin, everolimus, exatecan, exisulind, ferruginol, forode zine, fosfestrol, ICE chemotherapy regimen, IT-101, imexone, imiquimod, indolocarbazole, irofulven, laniquidar, larotaxel, lenalidomide, lucanton, lurtothecan, mafosfamide, mitozolomide, nafoxidine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw extract, pixantrone, proteasome inhibitor, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar, tesetaxel, triplatinum tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine , ZD6126 or zosuquidar.

The present invention further relates to a method of using compounds or pharmaceutical compositions provided herein in combination with radiation therapy to suppress abnormal cell growth or treat a hyperproliferative disorder in a mammal. Techniques for administering radiation therapy are known in the art, and these techniques can be used in the combination therapy described herein. Administration of a compound of the present invention in a given combination therapy can be determined as described herein.

Radiation therapy can be administered through one of several modalities or a combination of modalities, including, but not limited to, external beam radiation therapy, internal radiation therapy, implant-delivered radiation therapy, stereotactic radiosurgery, systemic radiation therapy, radiation therapy, and permanent or temporary interstitial brachytherapy. As used herein, the term brachytherapy refers to radiation therapy delivered by spatially confined radioactive material introduced into the body at or near a tumor or other site of proliferative tissue due to disease. The term is intended, without limitation, to include exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re186, Re-188, Sm-153, Bi-212, P-32 and Lu radioactive isotopes) . Suitable radiation sources for use as a cell enhancer of the present invention include both solids and liquids. As a non-limiting example, the radiation source may be a radionuclide such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation or other rays that have a therapeutic effect. The radioactive material may also be a fluid prepared from any solution of radionuclide(s), for example a solution of I-125 or I-131, or the radioactive fluid may be prepared using a slurry of a suitable fluid containing small particles of solid radionuclides such as like Au-198, Y-90. Moreover, radionuclide(s) may be included in the gel or radioactive microspheres.

The compounds or pharmaceutical compositions of the present invention can be used in combination with a certain amount of one or more substances selected from antiangiogenic agents, signal transduction inhibitors, antiproliferative agents, glycolysis inhibitors or autophagy inhibitors.

Antiangiogenic agents such as MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors and COX-11 (cyclooxygenase 11) inhibitors can be used in combination with the compound of the present invention and the pharmaceutical compositions described herein. document. Antiangiogenic agents include, for example, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib and rofecoxib. Examples of applicable

- 21 045678 matrix metalloproteinase inhibitors are described in WO 96/33172, WO 96/27583, European patent application publication EP 0818442, European patent application publication EP 1004578, WO 98/07697, WO 98/03516, WO 98/34918, WO 98/34915, WO 98/33768, WO 98/30566, European patent application publication 606046, European patent application publication 931788, wO 90/05719, WO 99/52910, WO 99/52889, WO 99/29667, WO 199900767 5 , European Patent Application Publication EP 1786785, European Patent Application Publication No. EP 1181017, US Patent Application Publication No. US20090012085, US Patent Application Publication US5863949, US Patent Application Publication US5861510 and European Patent Application 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 MMP-1 inhibitory activity. More preferred are inhibitors that selectively inhibit MMP-2 and/or AMP-9 over other matrix metalloproteinases (i.e. MAP-1, MMP-3, MMP-4, MMP5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12 and MMP-13). Some specific examples of MMP inhibitors useful in the present invention are AG-3340, RO 32-3555 and RS 13-0830.

The compounds of the present invention can also be used in combination therapy with other anticancer agents, such as acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim, arglabin, trioxide arsenic, BAM 002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine oxphosphate, DA 3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT-diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphate, exemestane, exisulind, fadrozole, filgrastim , finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamycin, gimeracil/oteracil/tegafur combination, glycopin, goserelin, heptaplatin, human chorionic gonadotropin, human germline alpha-fetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alpha, natural interferon alpha, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-N1, interferon alpha-„ ₃ , interferon alphacon-1, interferon-alpha, natural interferon-beta, interferon-beta 1a, interferon beta-1b, interferon-gamma, natural interferon gamma-1a, interferon gamma-1b, interleukin-1 beta, yobenguane, irinotecan, irsogladin, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon, leuprorelin, levamisole+fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, mirimostim, non-complementary double-stranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim, nafar elin, naloxone+pentazocine, nartograstim , nedaplatin, nilutamide, noscapine, new erythropoiesis-stimulating protein, octreotide NSC 631570, oprelvequin, osaterone, oxaliplatin, paclitaxel, pamidronic acid, pegaspargase, peginterferon-alpha-2b, sodium pentosan polysulfate, pentostatin, picibanil, pyrarubicin, antithymocyte rabbit polyclonal antibodies , polyethylene glycol interferon-alpha-2a, sodium porfimer, raloxifene, raltitrexed, razburicase, rhenium Re 186 etidronate, RII retinamide, rituximab, romurtide, samarium (153 Sm) lexidronam, sargramostim, sisofiran, sobuzoxan, sonermin, strontium-89 chloride, suramin , tazonermine, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropin alpha, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor alpha, natural, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulysin, zinostatin stimalamer, or zoledronic acid; abarelix; AE 941 (Aetema), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diaziquon, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, halocitabine, immunogen gastrin 17, HLA-B7 for gene therapy (Vical), granulocyte macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2, iproxifen, LDI 200 (Milkhaus), leridistim, lintuzumab, monoclonal antibody CA 125 (Biomira), anticancer monoclonal antibodies (Japan Pharmaceutical Development), monoclonal antibodies HER-2 and Fc (Medarex), idiotypic monoclonal antibody 105AD7 (CRC Technology), idiotypic monoclonal antibody CEA (Trilex), monoclonal antibody LYM-1-iodine 131 (Techniclone), monoclonal antibody to polymorphic epithelial mucin, yttrium 90 labeled (Antisoma), marimastat, menogaryl, mitumomab, motexafine gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, protein P 30, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, sparphosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN, now Pfizer, Inc.), TA 077 (Tanabe), tetrathiomolybdate, taliblastin, thrombopoietin, ethylethiopurpurine tin, tirapazamine, cancer vaccine (Biomira), anti-melanoma vaccine (New York University), anti-melanoma vaccine (Sloan-Kettering Institute), melanoma oncolysate vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital) or valspodar.

The compounds of the present invention can additionally be used with VEGFR inhibitors. Other compounds described in the following patents and patent applications can be used in combination therapy: US 6258812, US 2003/0105091, WO 01/37820, US 6235764, WO 01/32651, US 6630500, US 6515004, US 6713485, US 5521184 , US 5770599, US 5747498, 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/Wo 99/ 45009, WO 00/59509, WO 99/61422, US 5990141, WO 00/12089 and WO 00/02871.

In some embodiments, the combination includes a composition of the present invention in combination with at least one antiangiogenic agent. Such agents include, without limitation, chemical compositions produced in vitro synthetically, antibodies, antigen-binding sites, radionuclides, as well as their combinations and conjugates. The agent may be an agonist, antagonist, allosteric modulator, toxin or, more generally, may act in an inhibitory or stimulatory manner on its target (eg, activation or inhibition of a receptor or enzyme) and thereby promote cell death or arrest cell growth.

Exemplary antiangiogenic agents include ERBITUX™ (IMC-C225), KDR (kinase domain receptor) inhibitory agents (e.g., antibodies and antigen binding sites that specifically bind to the receptor kinase domain), anti-VEGF agents (e.g., antibodies and antigen-binding sites that specifically bind to VEGF, or soluble VEGF receptors or a ligand-binding site thereof), such as AVASTIN™ or VEGF-TRAP™, and VEGF receptor antibody agents (e.g., antibodies or antigen-binding sites that specifically bind to agents that inhibit EGFR (e.g., antibodies or antigen-binding sites that specifically bind to it), such as Vectibix (panitumumab), IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-Ang1 and Ang2 anti-EGFR agents (e.g., antibodies or antigen-binding sites that specifically bind to it or its receptors, e.g., Tie2/Tek), as well as inhibitory agents that are antibodies to Tie2 kinase (e.g., antibodies or antigen-binding sites that specifically bind to it ). The pharmaceutical compositions of the present invention may also contain one or more agents (for example, antibodies, antigen-binding sites or soluble receptors) that specifically bind to and inhibit the activity of growth factors, such as hepatocyte growth factor (HGF, also known as scattering factor) antagonists and antibodies or antigen-binding sites that specifically bind to its c-met receptor.

Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., US Patent Application Publication No. 2003/0162712, US Patent No. 6,413,932), anti-TWEAK antibody agents (eg, specifically binding antibodies or antigen binding sites or soluble receptor antagonists TWEAK, see Wiley, US patent No. 6727225), ADAM disintegrin domain as an antagonist of integrin binding to its ligands (Fanslow et al., publication of US patent application No. 2002/0042368), specifically binding anti-ephreceptor antibodies and/or anti-ephrin antibodies or antigen binding sites (US Pat. Nos. 5,981,245, 5,728,813, 5,969,110, 6,596,852, 6,232,447, 6,057,124 and members thereof) and antagonists that are anti-PDGF-BB antibodies (e.g., specific binding antibodies or antigen-binding sites), as well as antibodies or antigen-binding sites that specifically bind to PDGF-BB ligands, and agents that are inhibitors of PDGFR kinase (eg, antibodies or antigen-binding sites that specifically bind thereto).

Additional antiangiogenic/antitumor agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptanib octasodium (Gilead Sciences, USA); alfastatin (BioActa, UK); M-PGA (Celgene, USA, US 5712291); illomastat (Arriva, USA, US 5892112); emaxanib (Pfizer, USA, US 5792783); vatalanib (Novartis, Switzerland); 2methoxyestradiol (EntreMed, now CASI Pharmaceuticals, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); monoclonal antibody alpha-0148 (Amgen, USA); CEP7055,(Cephalon, USA); monoclonal antibody to Vn (Crucell, Netherlands) DAC:antiangiogen (ConjuChem, Canada); angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP 970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); fibrinogen fragment-E (BioActa, UK); angiogenesis inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); metastatin (EntreMed, USA); angiogenesis inhibitor (Tripep, Sweden); maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IVAX, USA); benefin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142

- 23 045678 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN), (Genentech, USA); angiogenesis inhibitors (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); monoclonal antibody, second generation alpha5beta3 integrin (Applied Molecular Evolution, USA and MedImmune, USA); a product for gene therapy of retinopathy (Oxford BioMedica, UK); enzastaurine hydrochloride (USAN) (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Institute for Research on Malignant Tumors of Genoa, Italy); angiogenesis inhibitor (Alchemia, Australia); VEGF antagonist (Regeneron, USA); antiangiogenic drugs based on rBPI 21 and BPI (XOMA, USA); PI 88 (Progen, Australia); cilengitide (pINN) (Merck KGaA, Germany; Technical University of Munich, Germany, Scripps Clinic and Research Foundation, USA); cetuximab (INN) (Aventis, France); AVE 8062 (Ajinomoto, Japan); AS 1404 (Oncology Research Laboratory, New Zealand); SG 292 (Telios, USA); endostatin (Boston Children's Hospital, USA); ATN 161 (Attenuon, USA); angiostatin (Boston Children's Hospital, USA); 2-methoxyestradiol (Boston Children's Hospital, USA); ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA); AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); vatalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); tissue factor signaling pathway inhibitors (EntreMed, USA); pegaptanib (Pinn) (Gilead Sciences, USA); xanthorizole (Yensei University, South Korea); genetically engineered vaccine, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX 103 (University of California at San Diego, USA); PX 478 (ProlX, USA); metastatin (EntreMed, now CASI Pharmaceuticals, USA); Troponin I (Harvard University, USA); SU 6668 (SUGEN, now Pfizer, Inc., USA); OXI 4503 (OXiGENE, USA); o-guanidines (Dimensional Pharmaceuticals, USA); motuporamine C (University of British Columbia, Canada); CDP 791 (Celltech Group, UK); atiprimod (pINN) (GlaxoSmithKline, UK); E 7820 (Eisai, Japan); CYC 381 (Harvard University, USA); AE 941 (Aeterna, Canada); angiogenesis vaccine (EntreMed, now CASI Pharmaceuticals, USA); urokinase plasminogen activator inhibitor (Dendreon, USA); oglufanide (pINN) (Melmotte, USA); HIF-1 alpha inhibitors (Xenova, UK); CEP 5214 (Cephalon, USA); BAY RES 2622 (Bayer, Germany); angiocidin (InKine, USA); A6 (Angstrom, USA); KR 31372 (Korea Chemical Technology Research Institute, South Korea); GW 2286 (GlaxoSmithKline, UK); EHT 0101 (ExonHit, France); CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); intraocular drug delivery system, 2methoxyestradiol (EntreMed, USA); Anginex (Maastricht University, the Netherlands, and the University of Minnesota, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); tumor necrosis factor alpha inhibitors (National Institute on Aging, USA); SU 11248 (Pfizer, USA and SUGEN USA); ABT 518 (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children's Hospital, USA and EntreMed, USA); monoclonal antibody, KDR (ImClone Systems, USA); monoclonal antibody, alpha5beta1 (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK, and Johnson & Johnson, USA); GFB 116 (University of South Florida, USA, and Yale University, USA); CS 706 (Sankyo, Japan); prodrug combretastatin A4 (Arizona State University, USA); chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan, and TAP, USA); AG 13925 (Agouron, USA); tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne University, USA) CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); monoclonal antibody, vascular endothelial growth factor (Xenova, 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 inhibitors (InSight, Israel); KL 3106 (Kolon, South Korea); honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); VEGF receptor modulators (USP); VE-cadherin-2 antagonists (ImClone Systems, USA); vasostatin (National Institute of Health, USA); Flk-1 vaccine (ImClone Systems, USA); TZ 93 (Tsumura, Japan); tumstatin (Beth Israel Hospital, USA); shortened soluble FLT 1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligands (Regeneron, USA) and thrombospondin 1 inhibitor (Allegheny Foundation for Health, Education and Research, USA).

Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR), algal toxins that inhibit autophagy that inhibit protein phosphatases type 2A or type 1, cAMP analogues, and drugs agents that increase cAMP levels such as adenosine, LY204002, N6mercaptopurine riboside and vinblastine. In addition, antisense or siRNA that inhibits the expression of proteins, including but not limited to ATG5 (which are involved in autophagy), can also be used.

- 24 045678

Additional pharmaceutically active compounds/agents that can be used in the treatment of cancers and that can be used in combination with one or more compounds of the present invention include epoetin alfa, darbepoetin alfa, panitumumab, pegfilgrastim, palifermin, filgrastim, denosumab, ancestim, AMG 102, AMG 176, AMG 386, AMG 479, AMG 655,

AMG 745, AMG 951 and AMG 706 or pharmaceutically acceptable salts thereof.

In some embodiments, a composition provided herein is administered in conjunction with a chemotherapeutic agent. Suitable chemotherapeutic agents may include natural products such as vinca alkaloids (eg, vinblastine, vincristine and vinorelbine), paclitaxel, epidipodophyllotoxins (eg, etoposide and teniposide), antibiotics (eg, dactinomycin (actinomycin D), daunorubicin, doxorubicin and idarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin), mitomycin, enzymes (e.g. L-asparaginase, which systemically metabolizes L-asparagine and deprives it of cells that do not have the ability to synthesize their own asparagine), antiplatelet agents, antiproliferative/antimitotic alkylating agents such as nitrogen mustards (eg, mechlorethamine, cyclophosphamide and analogues, melphalan and chlorambucil), ethyleneimines and methylmelamines (eg, hexamethylmelamine and thiotepa), CDK inhibitors (eg, seliciclib, UCN-01, P1446A-05, PD0332991, dinaciclib, P27-00, AT-7519, RGB286638 and SCH727965), alkyl sulfonates (eg, busulfan), nitrosourea derivatives (eg, carmustine (BCNU) and analogs, and streptozocin), trazenes-dacarbazinine (DTIC), antiproliferative/antimitotic antimetabolites such as folic acid analogs (eg, methotrexate), pyrimidine analogues (eg, fluorouracil, floxuridine and cytarabine), purine analogues and related inhibitors (eg, mercaptopurine, thioguanine, pentostatin and 2-chlorodeoxyadenosine), aromatase inhibitors (eg, anastrozole, exemestane and letrozole) and coordination platinum complexes (eg, cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (eg, trichostatin, sodium butyrate, apicidan, suberoylanilide hydroamic acid, vorinostat, LBH 589, romidepsin, ACY-1215 and panobinostat), mTor inhibitors (eg, temsirolimus, everolimus, ridaforolimus and sirolimus), KSP(Eg5) inhibitors (eg, Array 520), DNA binding agents (eg, Zalypsis), PI3K delta inhibitor (eg, GS-1101 and TGR-1202) , PI3K delta and gamma inhibitor (eg, CAL-130), multikinase inhibitor (eg, TG02 and sorafenib), hormones (eg, estrogen), and hormone agonists such as luteinizing hormone releasing factor (LHRH) agonists (eg, goserelin, leuprolide and triptorelin), BAFF neutralizing antibody (eg, LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 antibodies (eg, CNTO328), telomerase inhibitors (eg, GRN 163L), aurorakinase inhibitors (eg, MLN8237), monoclonal cell surface antibodies (eg, anti-CD38 (HUMAX-CD38), anti-CS1 antibodies (eg, elotuzumab), HSP90 inhibitors (eg, 17 AAG and KOS 953), P13K/Akt inhibitors (eg, perifosine), Akt inhibitor (eg , GSK-2141795), PKC inhibitors (eg, enzastaurine), FTI species (eg, Zarnestra™), anti-CD138 antibodies (eg, BT062), Torc1/2 specific kinase inhibitor (eg, INK128), kinase inhibitor (eg, GS -1101), ER/UPR targeting agent (eg, MKC-3946), cFMS inhibitor (eg, ARRY-382), JAK1/2 inhibitor (eg, CYT387), PARP inhibitor (eg, olaparib and veliparib (ABT-888) ), BCL-2 antagonist. Other chemotherapeutic agents may include mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or any analog or derivative of the above.

The compounds of the present invention can also be used in combination with radiation therapy, hormonal therapy, surgery and immunotherapy, such therapies being well known to those skilled in the art.

In some embodiments, a pharmaceutical composition provided herein is co-administered with a steroid. Suitable steroids may include, but are not limited to, acetoxypregnenolone, alklometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chlorprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivasol, deflazacort, desonide, deoxymethasone, dexamethasone, diflorazone, dif lucortolone, difuprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluopinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, galcinonide, halobetasol , halomethasone, hydrocortisone, loteprednol etabonate, mazipredone, medrisone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednilidene, rimexolone, thixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexace tonide, as well as their salts and/or derivatives. In a specific embodiment, the compounds of the present invention can also be used in combination with additional pharmaceutically active agents that treat nausea. Examples of agents that can be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron and prochlorpemazine or pharmaceutically acceptable salts thereof.

- 25 045678

The compounds of the present invention can also be used in combination with an additional pharmaceutically active compound that disrupts or inhibits the RAS-RAFERK or PI3K-AKT-TOR signaling pathways. In other such combinations, the additional pharmaceutically active compound is an antagonist of PD-1 and PD-L1. The compounds or pharmaceutical compositions of the present disclosure may also be used in combination with an amount of one or more substances selected from EGFR inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, Mcl-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasomes and immune therapeutics, including monoclonal antibodies, immunomodulatory imides (IMiDs), anti-PD-1, PDL-1, CTLA4, LAG1 and OX40 antibodies, GITR agonists, CAR-T cells and BiTE.

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

Non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications, and all pharmaceutically acceptable salts and solvates of these EGFR inhibitors: European patent application EP 520722, published December 30, 1992; European patent application EP 566226, published October 20, 1993; publication of PCT international application WO 96/33980, published on October 31, 1996; US Patent No. 5,747,498, issued May 5, 1998; publication of PCT international application WO 96/30347, published on October 3, 1996; European patent application EP 787772, published 6 August 1997; publication of PCT international application WO 97/30034, published on August 21, 1997; publication of PCT international application WO 97/30044, published on August 21, 1997; publication of PCT international application WO 97/38994, published on October 23, 1997; publication of PCT international application WO 97/49688, published on December 31, 1997; European patent application EP 837063, published April 22, 1998; publication of PCT international application WO 98/02434, published on January 22, 1998; publication of PCT international application WO 97/38983, published on October 23, 1997; publication of PCT international application WO 95/19774, published on July 27, 1995; publication of PCT international application WO 95/19970, published on July 27, 1995; publication of PCT international application WO 97/13771, published April 17, 1997; publication of PCT international application WO 98/02437, published on January 22, 1998; publication of PCT international application WO 98/02438, published on January 22, 1998; publication of PCT international application WO 97/32881, published on September 12, 1997; German patent application DE 19629652, published January 29, 1998; publication of PCT international application WO 98/33798, published on August 6, 1998; publication of PCT international application WO 97/32880, published on September 12, 1997; publication of PCT international application WO 97/32880, published on September 12, 1997; European patent application EP 682027, published November 15, 1995; publication of PCT international application WO 97/02266, published on January 23, 1997; publication of PCT international application WO 97/27199, published on July 31, 1997; publication of PCT international application WO 98/07726, published on February 26, 1998; publication of PCT international application WO 97/34895, published September 25, 1997; publication of PCT international application WO 96/31510', published on October 10, 1996; publication of PCT international application WO 98/14449, published April 9, 1998; publication of PCT international application WO 98/14450, published on April 9, 1998; publication of PCT international application WO 98/14451, published on April 9, 1998; publication of PCT international application WO 95/09847, published April 13, 1995; publication of PCT international application WO 97/19065, published on May 29, 1997; publication of PCT international application WO 98/17662, published April 30, 1998; US Patent No. 5,789,427, issued August 4, 1998; US Patent No. 5650415, issued July 22, 1997; US Patent No. 5656643, issued August 12, 1997; publication of PCT international application WO 99/35146, published on July 15, 1999; publication of PCT international application WO 99/35132, published on July 15, 1999; publication of PCT international application WO 99/07701, published on February 18, 1999; and publication of PCT international application WO 92/20642, published November 26, 1992. Additional non-limiting examples of small molecule EGFR inhibitors include any of the

- 26 045678 EGFR inhibitors, described in Traxler, P., 1998, Exp. Opin. Then Patents 8(12): 1599-1625.

Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block activation of EGFR by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi, H., et al., 1993, Br. J Cancer 67:247–253; Teramoto, T., et al., 1996, Cancer 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S. M, et al., 1999, Cancer Res. 15:59(8): 1935-40 and Yang, X., et al., 1999, Cancer Res. 59:1236–1243. Thus, the EGFR inhibitor may be a monoclonal antibody Mab E7.6.3 (Yang, 1999 supra) or Mab C225 (ATCC Accession No. HB8508) or an antibody or antibody fragment having binding specificity thereto.

The KRAS ^g12C inhibitors of the present invention can be used in combination with MEK inhibitors. Specific MEK inhibitors that can be used in combinations with the present invention include PD-325901, trametinib, pimasertib, MEK162 [also known as binimetinib], TAK-733, GDC-0973 and AZD8330. A particular MEK inhibitor that can be used together with a KRAS ^g12C inhibitor in the combinations of the present invention is trametinib (trade name: Mekinist®, commercially available from Novartis Pharmaceuticals Corp.). Another specific MEK inhibitor is N-(((2R)-2,3-dihydroxypropyl)oxy)-3,4-difluoro-2-((2-fluoro-4iodophenyl)amino)benzamide, also known as AMG 1009089, 1009089 or PD-325901. Another specific MEK inhibitor that can be used in the combinations of the present invention includes cobimetinib. MEK inhibitors include, but are not limited to, CI-1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, and ARRY-438162.

PI3K inhibitors include, but are not limited to, wortmannin, 17-hydroxywortmannin analogues described in WO 06/044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1yl]methyl]thieno [3,2-d]pyrimidin-4-yl]morpholine (also known as GDC 0941 and described in PCT Publications Nos. WO 09/036082 and WO 09/055730), 2-methyl-2-[4-[3-methyl -2-oxo-8-(quinolin-3-yl)-2,3dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235 and described in PCT- Publication No. WO 06/122806), (S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4morpholothieno[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- he, available from Axon Medchem), PI 103 hydrochloride (3-[4-(4-morpholinylpyrido-[3',2':4,5]furo[3,2d]pyrimidin-2-yl]phenol hydrochloride, available from Axon Medchem), PIK 75 (N'-[(1E)-(6bromimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfonohydrazide hydrochloride, available from Axon Medchem ), PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[1,2-c]quinazolin-5-yl)nicotinamide, available from Axon Medchem), GDC-0941 bismesylate (2-( 1H-Indazol-4-yl)-6-(4methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine bismesylate, available from Axon Medchem), AS-252424 (5 -[l-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-meth-(Z)-ylidene]thiazolidine-2,4-dione, available from Axon Medchem) and TGX-221 ( 7-methyl-2-(4-morpholinyl)-9-[1(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-one, available from Axon Medchem), XL-765 and XL -147. Other PI3K inhibitors include demethoxyviridine, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, palomide 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG1 00-115, CAL263, PI-103 , GNE-477, CUDC-907 and AEZS-136.

AKT inhibitors include, but are not limited to, Akt-1-l (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 (eg, Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-I-imidazo[4,5c]pyridinyl compounds (eg WO 05011700); indole-3-carbinol and derivatives thereof (eg, US Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134(12 Suppl), 3493S-3498S); perifosine (eg, interferes with membrane localization of Akt; Dasmahapatra et al. (2004) Clin. Cancer Res. 10(15), 524252, 2004); phosphatidylinositol ester, lipid analogues (eg Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); and triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al. (2004) Cancer Res. 64, 4394-9).

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 in the FKBP12 enhancer; rapamycins and their derivatives, including: CCI779 (temsirolimus), RAD001 (everolimus; WO 9409010) and AP23573; rapalogues, for example, as disclosed in WO 98/02441 and WO 01/14387, for example AP23573, AP23464 or AP23841; 40-(2hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also called CC1779), 40-epi-(tetrazolyte)-rapamycin (also called ABT578), 32-deoxorapamycin, 16pentynyloxy-32(S )-dihydrorapamycin and other derivatives disclosed in WO 05005434; derivatives disclosed in US Patent No. 5258389, WO 94/090101, WO 92/05179, US Patent No. 5118677, US Patent No. 5118678, US Patent No. 5100883, US Patent No. 5151413, US Patent No. 5120842, WO 93/111130, W O 94/02136, WO 94/02485, WO 95/14023, WO 94/02136, WO 95/16691, WO 96/41807, WO 96/41807 and US patent No. 5256790; phosphorus-containing rapamycin derivatives (eg WO 05016252); 4H-1-Benzopyran-4-one derivatives (e.g. US provisional patent application

- 27 045678 No. 60/528340).

MCl-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. Myeloid leukemia cell protein 1 (MCL-1) is one of the key anti-apoptotic members of the B cell lymphoma 2 (BCL-2) protein family. Overexpression of MCL-1 has been closely associated with tumor progression as well as resistance not only to traditional chemotherapeutic agents, but also to targeted therapeutics, including BCL-2 inhibitors such as ABT-263.

KRAS ^g12C inhibitors can also be used in combination with SHP2 inhibitors in the present invention. SHP2 inhibitors that can be used in combinations of the present invention include, but are not limited to, SHP099 and RMC-4550 or RMC-4630 from Revolutions Medicines in Redwood City, California.

Proteasome inhibitors include, but are not limited to, Kyprolis® (carfilzomib),

Velcade®(bortezomib) and oprosomib.

Immune therapeutic agents include, but are not limited to, anti-PD-1 antibody agents, anti-PDL-1 antibody agents, anti-CTLA-4 antibody agents, anti-LAG1 antibody agents, and anti-PDL-1 antibody agents. to OX40.

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

Immunomodulatory drugs (IMiDs) are a class of immunomodulatory drugs (drugs that regulate immune responses) containing an imide group. The IMiD class includes thalidomide and its analogues (lenalidomide, pomalidomide and apremilast).

Inhibitors directed against PD-1, including but not limited to antibodies, include but are not limited to pembrolizumab (Keytruda®) and nivolumab (Opdivo®). Exemplary anti-PD-1 antibodies and methods of use thereof are described in Goldberg et al., Blood 110(1):186-192 (2007), Thompson et al., Clin. Cancer Res. 13(6): 1757-1761 (2007) and Korman et al., International Application No. PCT/JP2006/309606 (Publication No. WO 2006/121168 A1), each of which is expressly incorporated by reference herein. Includes: Yervoy™ (ipilimumab) or tremelimumab (CTLA-4), galiximab (B7.1), BMS-936558 (PD-1), MK-3475 (PD-1), AMP224 (B7DC), BMS-936559 (to B7-H1), MPDL3280A (to B7-H1), MEDI-570 (to ICOS), AMG557 (to B7H2), MGA271 (to B7H3), IMP321 (to LAG-3), BMS-663513 ( anti-CD137), PF-05082566 (anti-CD137), CDX-1127 (anti-CD27), anti-OX40 (Providence Health Services), huMAbOX40L (anti-OX40L), atacicept (anti-TACI), CP-870893 (anti-CD40), lucatumumab (anti-CD40), dacetuzumab (anti-CD40), muromonab-CD3 (anti-CD3), ipilumumab (anti-CTLA-4). Immune therapeutics also include genetically engineered T cells (eg, CAR-T cells) and bispecific antibodies (eg, BiTE).

GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (eg, bivalent anti-GITR antibodies), such as the GITR fusion protein described in US Patent No. 6111090box.c, European Patent No. 090505B1, US Patent No. 8586023, PCT Publications No. No. WO 2010/003118 and 2011/090754, or an anti-GITR antibody described, for example, in US Patent No. 7025962, European Patent No. 1947183B1, US Patent No. 7812135, US Patent No. 8388967, US Patent No. 8591886, European Patent No. EP 1 866339 , 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, US Patent No. 7618632 and PCT Publication No. WO 2011 /051726.

The compounds described herein may be used in combination with the agents disclosed herein or other suitable agents depending on the condition being treated. Therefore, in some embodiments, one or more compounds of the present invention will be administered in conjunction with other agents described above. When used in combination therapy, the compounds described herein are administered simultaneously or separately with a second agent. This combination administration may include simultaneous administration of two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compound described herein and any of the agents described above can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compound of the present invention and any of the agents described above can be administered simultaneously, with both agents present in separate formulations. In another alternative, administration of a compound of the present invention may be immediately followed by administration of any of the agents described above, or vice versa. In some embodiments of a separate administration protocol, a compound of the present invention and any of the agents described above are administered several minutes apart, or several hours apart, or several days apart.

Whereas one aspect of the present invention provides for the treatment of a disease/conditions

- 28 045678 using a combination of pharmaceutically active compounds that can be administered separately, the present invention further relates to combining separate pharmaceutical compositions in the form of a kit. The kit contains two separate pharmaceutical compositions: a compound of the present invention and a second pharmaceutical compound. The kit contains a container for containing individual compositions, such as a divided bottle or a divided foil pouch. Additional examples of containers include syringes, boxes, and flexible reservoirs. In some embodiments, the kit contains instructions for use of the individual components. The kit form is particularly preferred when the individual components are preferably administered in different dosage forms (eg, orally and parenterally), administered at different dosage intervals, or when dosing of the individual components of the combination will be directed by the health care professional who prescribes the treatment.

All patents and other publications set forth herein are incorporated herein by reference.

The methods presented below illustrate specific embodiments of the present invention. Such methods are intended to be illustrative and are not intended to limit the scope of the claims in any way.

Suitable methods of the present invention.

The following intermediates related to 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl -4-(2-pronoul)-1-piperazunyl)nudo[2,3d]pyrimidin-2(III)-one are illustrative examples of the present invention and are not intended to be construed as limiting the scope of the present invention.

The synthesis of compound 9 and corresponding intermediates is described in US Patent Application Serial No. 15/984855, filed May 21, 2018, which claims priority and claims benefit from US Provisional Application No. 62/509629, filed May 22, 2017, which are incorporated herein by reference in their entirety for all purposes.

6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-( 2propenoyl)-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 isolated by chiral chromatography.

(1) (COCI) ₂ ,THF,75°C (2)

Intermediate S

Step 1: 2,6-dichloro-5-fluoronicotinamide (intermediate S). To a mixture of 2,6-dichloro-5fluoro-nicotinic acid (4.0 g, 19.1 mmol, AstaTech Inc., Bristol, PA) in dichloromethane (48 ml) was added oxalyl chloride (2 M solution in DCM, 11.9 ml , 23.8 mmol) followed by the addition of a catalytic amount of DMF (0.05 ml). The reaction mixture 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% as NH3, 3.6 mL, 28.6 mmol) was slowly added using a syringe. The resulting mixture was stirred at 0°C for 30 minutes and then concentrated. The residue was diluted with 1:1 EtOAc/heptane and shaken for 5 min, then filtered. The filtered solids were removed and the remaining stock solution was partially concentrated to half volume and filtered. The filtered solids were washed with heptane and dried in a reduced pressure oven (45°C) overnight to obtain

- 29 045678

2,6-dichloro-5-fluoronicotinamide.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm 8.23 (d, J=7.9 Hz, 1H) 8.09 (br s, 1H) 7.93 (br s, 1H); mass/charge (ESI, +ve ion): 210.9 (M+H) ⁺ .

Step 2: 2,6-dichloro-5-fluoro-N((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. To an ice-cooled slurry of 2,6-dichloro-5-fluoronicotinamide (intermediate S, 5.0 g, 23.9 mmol) in THF (20 mL) was added slowly oxalyl chloride (2 M solution in DCM, 14.4 mL, 28 .8 mmol) using a syringe. The resulting mixture was heated at 75°C for 1 hour, then the heating was stopped and the reaction mixture was concentrated to half the volume. After cooling to 0°C, THF (20 ml) was added followed by dropwise addition of a solution of 2-isopropyl-4-methylpyridin-3-amine (intermediate R, 3.59 g, 23.92 mmol) in THF (10 ml) using a cannula. The resulting mixture was stirred at 0°C for 1 hour and then quenched using a 1:1 mixture of saline and saturated aqueous ammonium chloride. The mixture was extracted with EtOAc (3x) and the combined organic layers were dried over anhydrous sodium sulfate and concentrated to give 2,6dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. This material was used in the next step without further purification. Mass/charge (ESI, +ve ion): 385.1 (M+H) ⁺ .

Step 3: 7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine2,4(1H,3H)-dione. To an ice-cooled solution of 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin3-yl)carbamoyl)nicotinamide (9.2 g, 24.0 mmol) in THF (40 mL) was added slowly KHMDS (1 M solution in THF, 50.2 ml, 50.2 mmol) using a syringe. The ice bath was removed and the resulting mixture was stirred for 40 minutes at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride and extracted with EtOAc (3x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-50% 3:1 EtOAc-EtOH/heptane) to give 7chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3 -d]pyrimidine-2,4(1H,3H)-dione.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 12.27 (br s, 1H), 8.48-8.55 (m, 2H), 7.29 (d, J=4.8 Hz, 1H ), 2.87 (quin, J=6.6 Hz, 1H), 1.99-2.06 (m, 3H), 1.09 (d, J=6.6 Hz, 3H), 1.01 (d, J=6.6 Hz, 3H). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -126.90 (s, 1F); mass/charge (ESI, +ve ion): 349.1 (M+H) ⁺ .

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]pyrimidine2,4(1H,3H)-dione (4.7 g, 13 .5 mmol) and DIPEA (3.5 mL, 20.2 mmol) in acetonitrile (20 mL), phosphorus oxychloride (1.63 mL, 17.5 mmol) was added dropwise using a syringe. The resulting mixture was heated at 80°C for 1 hour and then cooled to room temperature and concentrated to give 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 next step without further purification. Mass/charge (ESI, +ve ion): 367.1 (M+H) ⁺ .

Step 5: (S)-tert-butyl-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2dihydropyrido[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(1 H)-one (13.5 mmol) in acetonitrile (20 ml) was added DIPEA (7.1 ml, 40.3 mmol) followed by (S)-4N-Boc-2-methylpiperazine (3.23 g, 16.1 mmol, Combi-Blocks, Inc., San Diego, California, USA). The resulting mixture was warmed to room temperature and stirred for 1 hour, then diluted with cold saturated aqueous sodium bicarbonate (200 ml) and EtOAc (300 ml). The mixture was stirred for an additional 5 minutes, the layers were separated, and the aqueous layer was extracted with additional 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 give (S)-tert-butyl-4-(7-chloro-6-fluoro-1-(2-isopropyl-4methylpyridin-3-yl )-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1carboxylate. Mass/charge (ESI, +ve ion): 531.2 (M+H) ⁺ .

Step 6: (3 S)-tert-butyl-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1 -(2-isopropyl-4-methylpyridin3-yl)-2-oxo-1 ,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. Mixture of (S)tert-butyl-4-(7-chloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3d]pyrimidine -4-yl)-3-methylpiperazine-1-carboxylate (4.3 g, 8.1 mmol), potassium trifluoro(2-fluoro-6hydroxyphenyl)borate (intermediate Q, 2.9 g, 10.5 mmol) , potassium acetate (3.2 g, 32.4 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(P), 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 hour. Allow the reaction mixture to cool to room temperature, quench it with semi-saturated aqueous sodium bicarbonate and extract with EtOAc (2x) and DCM (1x) . The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-60% 3:1 EtOAc-EtOH/heptane) to give (3S)tert-butyl-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)- 1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2- 30 045678 dihydropyrido[2,3-d]nirimidin-4-yl)-3-methylpiperazin-1-carboxylate.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm 10.19 (br s, 1H), 8.38 (d, J=5.0 Hz, 1H), 8.26 (dd, J=12.5 , 9.2 Hz,

1H), 7.23-7.28 (m, 1H), 7.18 (d, J=5.0 Hz, 1H), 6.72 (d, J=8.0 Hz, 1H), 6, 68 (t, J=8.9 Hz, 1H), 4.77-4.98 (m,

1H), 4.24 (br t, J=14.2 Hz, 1H), 3.93-4.08 (m, 1H), 3.84 (br d, J=12.9 Hz, 1H), 3.52-3.75 (m, 1H), 3.07-3.28 (m, 1H), 2.62-2.74 (m, 1H), 1.86-1.93 (m, 3H ), 1.43-1.48 (m, 9H), 1.35 (dd, J=10.8, 6.8 Hz, 3H), 1.26-1.32 (m, 1H), 1, 07 (dd, J=6.6, 1.7 Hz, 3H), 0.93 (dd, J=6.6, 2.1 Hz, 3H). ¹⁹ F NMR (376 MHz, DMSO-d ₆ ) δ: -115.65 (s, 1F), -128.62 (s, 1F); mass/charge (ESI, +ve ion): 607.3 (M+H)+.

Step 7: 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(4-methyl-2-(2-propanyl)-3-pyridinyl)-4-((28)-2methyl-4- (2-nropenoyl)-1-niperazinyl)pyrido[2,3-d]nirimidin-2(1H)-one. Trifluoroacetic acid (25 ml, 324 mmol) was added to a solution of (38)-tert-butyl-4-(6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-(2isonronyl-4-methylpyridin-3- yl)-2-oxo-1,2-dihydronirido[2,3-d]pyrimidin-4-yl)-3-methylniperazine 1-carboxylate (6.3 g, 10.4 mmol) in DCM (30 ml). The resulting mixture was stirred at room temperature for 1 hour and then concentrated. The residue was dissolved in DCM (30 ml), cooled to 0°C and treated sequentially with DIPEA (7.3 ml, 41.7 mmol) and a solution of acryloyl chloride (0.849 ml, 10.4 mmol) in DCM (3 ml; added drop by drop using a syringe). The reaction mixture was stirred at 0°C for 10 min, then quenched with semi-saturated aqueous sodium bicarbonate and extracted with DCM (2x). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-100% 3:1 EtOAc-EtOH/heptane) to give 6-fluoro-7-(2-fluoro-6hydroxyphenyl)-1-(4-methyl-2-(2- propanyl)-3-pyridinyl)-4-((2S)-2-methyl-4-(2-propenoyl)-1πipiperazinyl)pyrido[2,3-d]pyrimidin-2(1H)-one.

^1H NMR (400 MHz, DMSO-d6) δ ppm 10.20 (s, 1H), 8.39 (d, J=4.8 Hz, 1H), 8.24-8.34 (m, 1H) , 7.23-7.32 (m, 1H), 7.19 (d, J=5.0 Hz, 1H), 6.87 (td, J=16.3, 11.0 Hz, 1H), 6.74 (d, J=8.6 Hz, 1H), 6.69 (t, J=8.6 Hz, 1H), 6.21 (br d, J=16.2 Hz, 1H), 5 .74-5.80 (m, 1H), 4.91 (br s, 1H), 4.23-4.45 (m, 2H), 3.97-4.21 (m, 1H), 3. 44-3.79 (m, 2H), 3.11-3.31 (m, 1H), 2.67-2.77 (m, 1H), 1.91 (s, 3H), 1.35 ( d, J=6.8 Hz, 3H), 1.08 (d, J=6.6 Hz, 3H), 0.94 (d, J=6.8 Hz, 3H). ¹⁹ F NMR (376 MHz, DMSO-d6) δ ppm -115.64 (s, 1F), -128.63 (s, 1F); mass/charge (ESI, +ve ion): 561.2 (M+H)+.

The present invention provides the following steps, where the separation of the rac-dione in steps 4 and 5 provides successful separation of atropisomers:

Stage 2

Stage 1

Stage 3 α

CI o

Rac-dione Yield 41% (2 stages) 4

CI with reverse

2-MeTHF, 0°C

NaOtBu (2.5 eq.)

i. (+)-DBTA (2 eq.)

ii. NH4OH

2-MeTHE/hsptan (7:5, 12 vol.) from 75°C to room temperature.

Pipayulin

Yield 58%

Stage 5 ii. Na2HPO4 (aq) MTBE

C.I.

DCM, DMF(KaT.) ---------- _but

Stage 4

Yield 39% (2 stages)

Stage 6 i. POCL, DIPEA toluene

MeO

Stage A2

TFA, DCM

Stage 8

Vos.

Des-boc Yield 94%

Stage 7

Cl ₂ Pd(dppD (2.0%) KOAc, dioxane

VVgz, DCM

Stage 9

Stage 10 sg

i. EUN/water AcOH

EtjN, NMP о°с

Biaryl Yield 80%

BUT

KF _3B

Boronite Yield 86% ii. Pin mill

Connection 9

Yield 80%

Stage A2

KF citric acid 1 ------------(BUT) ₂ V

Stage A1

i. i-BuLi, diisopropidamine ii. (EtO^B iii. Hydrochloric acid

BUT

Boronic acid

- 31 045678

Description of an illustrative method of the present invention. Stage 1.

F 1. (COCI) ₂ F NO /H NMSCHON . H ₂ N ^X ^C1 CH ₂ C1 ₂ CI O / ⁼ Ν o )=N Cl ₁ Cl ₁ Acid Amide CAS 82671 -06-5 CAS 113237-20-0 Material CAS No. MW (g/mol) Equivalent volumes Moli Theoretical quantity 2,6-Dichloro-5-fluoro-3pyridinecarboxylic acid 82671-06-5 209.99 1.0 eq. 119.1 25 kg DCM 74-09-2 84.93 16.51 eq. 2354, 200 kg 9 DMF 68-12-2 73.09 0.068 eq. 8.1 592 g (627 ml) Oxalyl chloride 79-37-8 126.93 1.25 eq. 148.9 18.9 kg Ammonium hydroxide 1336-21-6 35.05 5 eq. 595.5 40.2 l Water 7732-18-5 18.02 n. d. n. d. 261 l

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 and 20°C. Additional dichloromethane (33 kg) was added as a wash and the reaction mixture was stirred for 2 hours. The reaction mixture was cooled, then quenched with ammonium hydroxide (40.2 L; 595.5 mol) while maintaining the internal temperature at 0 ± 10°C. The resulting slurry was stirred for 90 minutes, then the product was collected by filtration. The filtered solids were washed using deionized water (3X 87 L) and dried to obtain 2,6-dichloro-5-fluoronicotinamide (compound 2).

Stage 2.

i. (coci) ₂ ,

CAS 113237-20-0 Aniline

CAS 1698293-93-4

Material CAS No. MW (g/mol) Equivalents/ volumes Moli Theoretical quantity Amide (2,6-dichloro-5fluoronicotinamide) 113237- 20-0 209.99 1.0 eq. 77.8 16.27 kg Oxalyl chloride 79-37-8 126.93 1.2 eq. 93.8 11.9 kg (7.9 l) Dichloromethane 75-09-2 84.93 n. d. n. d. 730.7 kg (551.5 l) DCM aniline solution, 169829 3-93-4 150.22 1.1 eq. 85.9 12.9 kg (weight containing representing Xia aniline) about myself 2-isopropyl-4- methylpyridine- 3-amine

In reactor A, oxalyl chloride (11.9 kg; 93 .8 mol) while maintaining the temperature at < 25°C for 75 min. The resulting solution was then heated to 40°C ± 3°C and held for 3 hours. The solution was distilled using vacuum 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-3amine (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 at 20 to 25°, displacing with additional dichloromethane until the solution was dry by KF analysis (limit < 0.05%). The solution was brought to a volume of approximately 23 L using dichloromethane. The dried aniline solution was then added to reactor A while maintaining the internal temperature at 0 ± 5°C throughout the addition procedure. The mixture was then heated to 23°C and held for 1 hour. The solution was finally filtered into a clean reactor to give 2,6-dichloro-5-fluoro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl) nicotinamide (compound 3) as a solution in DCM and used directly in the next step. Stage 3.

Gae-Dion 4

Material No. CAS MW (g/mol) Equivalents/volumes Like and Theoretical quantity Urea, solution in DCM 2,6-dichloro-5-fluoro-A{[4-methyl-2-(propan2-yl)pyridin-3yl]carbamoyl}pyridinium n-3-carboxamide n. d. 385.22 1.0 eq. 38.9 208.3 kg (15 kg contained weight) 2- Methyltetrahydrofur and 96-47- 9 86.13 n. d. n. d. 308 kg (358 l) Sodium tert-butoxide 865- 48-5 96.11 2.0 eq. 97.8 9.4 kg Ammonium chloride 12125- 02-9 53.49 n. d. 430 23.0 kg Hydrochloric acid 7467- 01-0 36.46 n. d. 41 1.6 kg Magnesium sulfate 7487- 88-9 120.37 n. d. 195 23.5 kg Sodium chloride 7647- 14-5 58.44 n. d. 282 16.5 kg Heptane 142- 82-5 100.21 n. d. n. d. 94L 10% citric acid 75 kg

In a solution of 2,6-dichloro-5-fluoro-N-{[4-methyl-2-(nropan-2-yl)pyridin-3-yl]carbamoyl}pyridin-3carboxamide in dichloromethane (urea (compound 3)) ( 15 kg contained weight; 38.9 mol) solvent exchanged to 2-MeTHF using vacuum distillation while maintaining internal temperature at 20-25°C. The reactor volume was adjusted to 40 L and then additional 2-MeTHF (105.4 kg) was charged. Sodium tert-butoxide (9.4 kg; 97.8 mol) was added while maintaining 510°C. The contents were heated to 23°C and stirred for 3 hours. The contents were then cooled to 05C and ammonium chloride (23.0 kg; 430 mol) was added as a solution in 60 L of deionized water. The mixture was heated to 20 C, and deionized water (15 l) was added, and further kept in

- 33 045678 within 30 minutes. Stirring was stopped and the layers were separated. The aqueous layer was removed and deionized water (81.7 L) was added to the organic layer. A mixture of conc. HCl (1.5 kg) and water (9 L), then slowly added to the reactor until the measured pH value reached 4-5. The layers were separated and the aqueous layer was back-extracted with 2-MeTHF (42.2 kg). The two organic layers were combined and washed with 10% citric acid (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), repeating as necessary until the target aqueous solution pH was >7.0. The organic layer was washed again with brine (54.7 kg) and then dried over magnesium sulfate (5 kg). The mixture was filtered to remove magnesium sulfate, washing the filter layer with 2-MeTHF (49.2 kg). The combined filtrate and washings were distilled using vacuum to a volume of 40 L. The concentrated solution was heated to 55°C and heptane (10-12 kg) was added slowly until the cloud point was reached. The solution was cooled to 23°C over 2 hours, then heptane (27.3 kg) was added over 2 hours. The product slurry was kept for 3 hours at 20-25°C, 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 the solid 7-chloro-6-fluoro-1-(2-isopropyl-4methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2,4(1H,3H)- dione (rac-dione (compound 4)).

Stage 4.

Cocrystal of M-dione with DBTA

Material CAS No. MW (g/mol) Equivalents/volumes Moli Theoretical quantity Rac-dione n. d. 348.76 1.0 (+)-2,3-Dibenzoyl-D- wine acid 17026-42-5 358.30 2.0 2-Methyltetrahydrofuran 96-47-9 86.13 7.0 Heptane 142-82-5 100.21 2.0 Heptane 142-82-5 100.21 h,o 2-Methyltetrahydrofuran 96-47-9 86.13 4.0 Heptane 142-82-5 100.21 2.0

(+)-2,3-Dibenzoyl-D-tartaric acid (2.0 equiv) was added to a vessel containing a stirred suspension of compound 4 (1.0 eq.) in 2-methyltetrahydrofuran (7.0 L/kg) under atmosphere. nitrogen. 2-MeTHF is chiral, but is used as a racemic mixture. Different enantiomers of 2-MeTHF are randomly included in the cocrystal. The resulting suspension was heated to 75°C and kept at 75°C until complete dissolution was observed (no more than 30 min). The resulting solution was finally filtered at 75°C into a second vessel. The final filtered solution was charged with n-heptane (2.0 L/kg) at a rate that maintained the internal temperature above 65°C. The solution was then cooled to 60°C, seeded with crystals (0.01 kg/kg) and left to settle for 30 minutes. The resulting suspension was cooled to 20°C for 4 h, and then samples were taken for chiral purity analysis by HPLC. Nheptane (3.0 L/kg) was loaded into the suspension, and then it was kept for 4 hours at 20°C in a nitrogen atmosphere. The suspension was filtered and the isolated solids were washed twice with (2:1) n-heptane:2methyltetrahydrofuran (3.0 L/kg). The material was dried using nitrogen and vacuum to obtain the M-dione:DBTA:Me-THF complex (compound 4a).

- 34 045678

Stage 5.

M-duon/DBTA/2-MeTHF 4a cocrystal

Material CAS No. MW (g/mol) Equivalents/volumes Like and Theoretical quantity Cocrystal M- dione/DBTA/Me-THF n. d. 1228.08 1.0 74.2 46.9 kg (25.9 kg including corrections for Mdion) Methyl Shresh Butyl Ether 1634-04-4 88.15 45.0 17593 2100 l Disodium hydrogen phosphate 7558-79-4 141.96 2.0 148.4 21.1 kg Purified water grade Where necessary USP quantity Magnesium sulfate 7487-88-9 120.37 n. d. n. d. 25 kg Heptane 142-82-5 100.20 60.0 19322 2835 l

In vessel A, a suspension of disodium hydrogen phosphate (21.1 kg, 2.0 eq) in deionized water (296.8 L, 6.3 L/kg) was stirred until dissolution was observed (>30 min). In vessel B, a suspension of M-dione complex:DBTA: Containing Me-THF complex (composition 4a) [46.9 kg (25.9 kg, corrected for M-dione, 1.0 eq.)] in methyl tert -butyl ether (517.8 L, 11.0 L/kg) was stirred for 15-30 minutes. The resulting solution from vessel A was added to vessel B and the mixture was then stirred for over 3 hours. Stirring was stopped and the two-phase mixture was allowed to separate for over 30 minutes. The lower aqueous phase was removed and then back-extracted with methyl tert-butyl ether (77.7 L, 1.7 L/kg). The organic phases were combined in vessel B and dried over magnesium sulfate (24.8 kg, 0.529 kg/kg). The resulting suspension from Vessel B was stirred for over three hours and then filtered into Vessel C. Vessel B was charged with methyl tert-butyl ether wash (46.9 L, 1.0 L/kg) and then filtered into vessel C. The contents of vessel C were cooled to 10°C and then distilled under vacuum while heating slowly to 35°C. Distillation was continued until 320-350 kg (6.8-7.5 kg/kg) of methyl tert-butyl ether was collected. After cooling the contents of vessel C to 20°C, n-heptane (278.7 L, 5.9 L/kg) was charged over one hour and then distilled under vacuum while heating slowly to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg/kg) of a mixture of methyl tert-butyl ether and n-heptane was collected. After cooling the contents of vessel C to 20°C, n-heptane (278.7 L, 5.9 L/kg) was charged a second time within one hour and then distilled under vacuum while heating slowly to 35°C. Distillation was continued until 190-200 kg (4.1-4.3 kg/kg) of a mixture of methyl tert-butyl ether and n-heptane was collected. After cooling the contents of vessel C to 20°C, n-heptane (195.9 L, 4.2 L/kg) was charged a third time within one hour, and then a sample was taken for solvent composition analysis by GC. The suspension of vessel C continued to stir for more than one hour. The suspension was filtered and then washed with n-heptane wash (68.6 L, 1.5 L/kg) from vessel C. The isolated solids were dried at 50°C and the sample was submitted for testing against starting material. Compound 5M was obtained, which was 7-chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3yl]pyrido[2,3-d]pyrimidine-2, 4(1H,3H)-dione (M-dione).

The first generation method described above was successfully scaled up to 200+ kg of rac-dione starting material (compound 5). In this method, seeding crystallization with a thermodynamically stable crystalline form of the rac-dione (which exhibits low solubility) can lead to

- 35 045678 for the defective batch. Based on subsequent studies, the present inventors have discovered that increasing DBTA equivalents and decreasing seed temperature by adjusting the heptane loading schedule improves process reliability. The improved method is robust to the presence of the thermodynamically stable crystalline form of the rac-dione and facilitates successful separation of atropisomers. Subsequent batches will include an improved process for large-scale production.

Stage 6.

Vos

Material CAS No. MW (g/mol) Equivalents/volumes Like and Theoretical quantity M-dion And. d. 348.76 1 eq. 9.8 3.7 kg Toluene 108-88-3 92.14 And. d. 375 34.6 kg (40 l) Phosphoryl chloride 10025-87-3 153.33 1.2 eq. I,7 1.8 kg (1.1 l) Ν,Ν- Diisopropylethylamine 7087-68-5 129.24 3.0 eq. 29.4 3.8 kg (5.1 l) (s)-1 -Sun-3 - methylpiperazine 147081-29-6 200.28 1.1 eq. 10.8 2.214 kg Bicarbonate of soda 144-55-8 84.01 And. d. And. d. 973 g Dichloromethane 75-09-2 84.93 And. d. 871 74 kg (55.6 l) Sodium chloride 7647-14-5 58.44 And. d. 103 6.0 kg Ethyl acetate 141-78-6 88.11 And. d. 288 25.4 kg (28.2 l)

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 (M-dione) (3.7 kg; 9.8 mol) was combined in reactor (A) with 10.5 kg of toluene and distilled to remove water to an oil, while maintaining the set temperature at 45° C. Toluene (21 kg) was added to the residue and the mixture was stirred for 30 minutes at 40-45°C. The contents were cooled to 22°C, then phosphoryl chloride (1.8 kg; 11.7 mol) was added. The mixture was cooled to 0-5°C before adding N,N-diisopropylethylamine (2.5 kg; 19.34 mol), while maintaining the temperature at <5°C. The solution was kept for 3 hours at 22°C. 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 loaded into reactor (A), while maintaining the temperature at < 25°C. The reaction mixture was kept for 15 min at 22°C, then quenched using sodium bicarbonate (973 g) in water (12.9 L), while maintaining the temperature at < 25 C. The mixture was stirred for 30 min, then added DCM (36.8 kg) while stirring continued for 1 hour. The layers were separated and the lower organic layer was poured into reactor (C). The aqueous layer in reactor (A) was back-extracted with DCM (18.4 kg) and the combined organic layers were washed with brine (6.0 kg NaCl; 16.5 kg deionized water). The organic layer was distilled at atmospheric pressure while maintaining the internal temperature at 45 to 55°C. DCM was replaced during distillation to azeotropically dry the solution. After distillation, the volume of the solution was brought to 19 L using DCM. The solution was cooled to 30°C and finally filtered. The filtrate was combined with ethyl acetate (8.5 kg) and then distilled at atmospheric pressure to collect 11-13 kg in a receiving vessel. The solution was seeded with 30 g of the original product and kept for 1 hour at 25-30°C, then further distilled at atmospheric pressure at an internal temperature of 45-55°C until 8.2 kg of distillate was collected. The suspension was cooled to 22°C and kept overnight, then further cooled to 0-5°C. The product was collected by filtration and washed twice with ethyl acetate (4.2 kg each time). The filter cake was dried under nitrogen and vacuum to give tert-butyl-(3S)-4-{7-chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2yl)pyridin- 3-yl]-2-oxo-1,2-dihydropyrido[2,3-b]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate (compound 6, pipezoline).

- 36 045678

Stage 7.

Cl ₂ Pd(dppf) (2.0%) KO Ac, dioxane

KF ₃ B

F

Boronat

Material CAS No. MW (g/mol) Equivalents/volumes Moli Theoretical quantity [1.1’- Bis(diphenylphosphino)ferrocene]dichloropalladium(P) 72287- 26-4 731.714 0.020 1.01 0.74 kg Dichloromethane 75-09-2 84.93 n. d. 400 kg 1,4-Dioxane 123-91-1 88.1052 5.0 n. d. 168 kg Ethylenediaminetetraacetic acid disodium salt dihydrate 6381-92- 6 336.207 1.0 45.2 15.2 kg Heptane 142-82-5 100.21 200 kg Nitrogen If necessary PTO quantity Pipazoline n. d. 531.0 1.0 45.2 24.0 kg Potassium acetate 127-08-2 98.1417 5.0 225.99 22.2 kg Trifluoro(2-fluoro-6- Potassium hydroxyphenyl borate n. d. 233.03 1.20 54.24 12.6 kg 2-Propanol 67-63-0 66.10 n. d. 850 kg Si-thiol n. d. n. d. n. d. 13.2 kg Sodium hydroxide 1310-73- 2 40.00 6.5 kg USP Grade Purified Water If necessary PTO quantity

Degassed dioxane (74.2 kg), tert-butyl (38)-4-{7-chloro-6-fluoro-(1M)-1[4-methyl-2-(propan-2-yl)pyridine was added to the reactor -3-yl]-2-oxo-1,2-dihydropyrido[2,3-pyrimidin-4-yl}-3methylpiperazine-1-carboxylate (compound 6, pipazoline) (24.0 kg, 45.2 mol ), potassium acetate (22.2 kg, 45.2 mol) and (dppf)PdCl ₂ (0.74 kg, 1.01 mol). An inert atmosphere was created in the reactor using nitrogen gas. The solution was purged with nitrogen gas until the oxygen content was <500 mg/L. The reaction mixture was heated to 87.5°C. A solution of potassium trifluoro(2-fluoro-6hydroxyphenyl)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 reaction mixture, maintaining the internal temperature at 82.5°C ± 7.5°C. The reaction mixture was brought to 87.5°С ± 1.5°С 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), maintaining the internal temperature at 85 °C ± 5 °C. The reaction mixture was cooled to 20°C ± 3°C over >2 hours and then stirred for >16 hours. The reaction mixture was filtered and the crude solids were washed with water (3x120 kg). The solids were washed with a mixture of heptane (28.8 kg) and 2-propanol (33.1 kg) and then dried at <50°C for >10 hours. The clean reactor was charged with crude solids and dichloromethane (240 kg). The contents were stirred at 20°C

-37045678 ± 5°C for more than 30 min. Si-thiol (144 kg) and dichloromethane (14.9 kg) were added to the reactor. The reaction mixture was stirred at 20°C ± 5°C for 18 hours. The reaction mixture was filtered and washed with dichloromethane (84 kg). The solution was distilled and the solvent was replaced with 2-propanol. The reaction mixture was heated to 60°C ± 3°C and heptane (108 kg) was charged while maintaining the temperature of the reaction mixture at 60°C ± 3°C. The reaction mixture was stirred for 45 minutes and then cooled and stirred at 20°C ± 5°C for 2.5 hours. The reaction mixture was filtered and washed with 50% v/v. heptane/2-propanol (61.9 kg). The isolated solids were dried at <50°C for >12 h to give tert-butyl-(3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[ 4-methyl-2-(propan-2yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazin-1-carboxylate (compound 7, biaryl).

Stage 8.

Biaryl 7 Des-BOC 8

General note. All equivalents and volumes are relative to biaryl 7.

Material CAS No. MW (g/mol) Equivalents/volumes Moli Theoretical quantity Biaryl 7 n. d. 606.67 1.0 eq. 5.27 2.75 kg T.F.A. 76-05-1 114.02 11 eq. 49.7 5.67 kg DCM 74-09-2 84.93 5 rev. n. d. 13.71 l Methanol 67-56-1 32.04 5 rev. n. d. 13.71 l Water 7732-18- 5 18.02 20 rev. n. d. 54.8 l Carbonate potassium 584-08-7 138.20 18 eq. 94.91 11.24 kg DCM 74-09-2 84.93 1 rev. n. d. 2.75 l Water 7732-18- 5 18.02 10 rev. n. d. 27.5 l Water 7732-18- 5 18.02 10 rev. n. d. 27.5 l

Tert-butyl-(3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2(propan-2-yl)pyridine was added to the reactor -3-yl]-2-oxo-1,2-dihydropyrido[2,3-pyrimidin-4-yl}-3-methylpiperazine-1carboxylate (compound 7, biaryl) (2.75 kg, 5.27 mol), DCM ( 13.7 L) and TFA (5.67 kg, 49.7 mol). The reaction mixture was stirred for 8-16 hours at 20 ± 5°C. Potassium carbonate (11.24 kg), water (54.8 L) and methanol (13.7 L) were added to the second reactor to form a homogeneous solution. The reaction mixture was added to the potassium carbonate solution over 2 hours. The mixture was stirred at 20 ± 5°C for an additional 12 hours. The resulting slurry was filtered and washed with water (2x27.5 L). The wet filter cake was dried for 24 hours to obtain 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2methylpiperazin-1 -yl]-(1M)-1-[4-methyl -2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-b]pyrimidin-2(1H)one (compound 8, DESBOC).

- 38 045678

Stage 9.

Des-BOC 8 iii. Heating cycle i'Pr iv. Filtration, water washing

ABOUT

JJ 1.3 eq. ί.

about. NMP, 2 hours

y. Extinguishing water Να ₂ ΗΡΟ ₄

Ch ¹

Crude compound 9

General note. All equivalents and volumes are relative to Des-BOC

Material CAS No. MW (g/mol) Equivalents/ volumes mmol Weight Volume Des-BOS n. d. 506.56 1.0 eq. 308.4 156.25g Acryloyl chlorideK 1 814-68-6 90.51 1.3 eq. 401.0 36.29g - - N.M.P. N- methylpyrrolidinone N2 872-50-4 99.13 4 rev. n. d. - 625 ml Water 7732-18- 5 18.02 20 rev. n. d. 3125g 3125 ml _Na2HPO4N3 7558-79- 4 141.96 4 eq. 1233.6 175.12g - Water 7732-18- 5 18.02 20 rev. n. d. 3125g 3.125 ml

6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(28)-2-methylpiperazin-1-yl]-(1M)-1-[4-methyl-2-(propan-2yl) Pyridin-3-yl]pyrido[2,3-b]pyrimidin-2(1H)-one (compound 8, DESBOC) (156.25 g) was combined with Nmethylpyrrolidinone (625 ml) and stirred at ambient temperature. Acryloyl chloride (36.29 g, 401.0 mmol) was added to the resulting solution while maintaining the internal temperature at <30°C. The contents were stirred for 2 hours at 25°C. In a separate reactor, a solution of sodium hydrogen phosphate (175.1 g; 1234 mmol) in deionized water (3.1 L) was prepared. The crude product solution was then transferred to a reactor containing sodium hydrogen phosphate solution for >2 hours at 25°C. The slurry was heated to 45°C in the middle of the addition procedure and after completion of the addition it was kept for 2 hours at the same temperature. The mixture was cooled to 25°C and held for 4 hours before collecting solids by vacuum filtration. The solids were 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-[(28)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[ 2,3-b]pyrimidin-2(1H)-one (crude compound 9).N4

Stage 10.

i. Ethanol/water 4:1 (9.4 vol. 1.5 eq. acetic acid, heating

gRg

Crude compound 9 ii. Final filtration iii. Adding water (15.5 vol) iv. Filtration, washing with ethanol/water o m iPr—l

Connection 9

General note. All equivalents and volumes are relative to the crude drug substance.

-39045678

Material CAS No. MW (g/mol) Equivalents/volumes mmol Weight Obye m Unrefined n. d. 560.60 1.0 eq. 253.9 142.33 g connection 9 Ethanol (200 proof) 64-17-5 - 7.5 rev. - - 1067 ml Water class USP - 18.02 1.9 rev. - - 270 ml Vinegar 64-19-7 60.05 1.5 eq. 380.8 22.87 g 21.82 acid ml Water class WFI - 18.02 15.5 rev. - - 2200 ml Ethanol (for rinsing) 64-17-5 - 2.5 rev. - - 356 ml Water class WFI (for flushing) - - 5.0 rev. - - 712 ml Seed in form 560.60 0 0.3-0.7 g connections 9N5

6-Fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-4-[(2S)- 2Methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (crude 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 finally filtered into a clean reactor. The solution was cooled to 45°C and then water (1067 ml) was added while maintaining the internal temperature at >40°C. The solution was seeded with starting compound 9, and the resulting mixture was kept for 30 min. Water (1138 ml) was then added over 2 hours. The mixture was cooled to 25°C and held for 8 hours, after which 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 give 6-fluoro-7-(2-fluoro-6hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl] -4-[(2S)-2-methyl-4-(prop-2enoyl)piperazin-1-yl]pyrido[2,3-d]pyrimidin-2(1H)-one (compound 9).

Reaction scheme and loading table for step A1.

i. n-BuLi, diisopropylamine iLBCEtOL_____________ iii. HCl (aq)

3-Fluoroanisole (2-Fluoro-6-methoxyphenyl)boronic acid

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Material CAS No. MW (g/mol) Equivalents/volumes mole Weight (G) Volume (l) Z-Fluoroanisole 456-49-5 126.13 1.0 1.19 150 0.136 n-Butyllithium (2.5 M in hexane) 109-72-8 64.06 1.5 1.78 And. d. 0.712 Diisopropyl min 108-18-9 101.19 1.4 1.66 168 0.233 Triethyl borate 150-46-9 145.99 2.0 2.38 347.5 0.405 Tetrahydrofran 109-99-9 72.11 12 rev. n. d. n. d. 1.8 Hydrochloric acid (2 and.) 7647-0Ι- Ο 36.46 10 rev. And. d. And. d. 1.5 Methyl tert-butyl ether 1634-04- 4 88.15 12 rev. n. d. n. d. 1.8 Heptane 142-82-5 100.20 10.5 rev. And. d. And. d. 1.575

Reactor A was charged with THF (6 vol) and diisopropylamine (1.4 eq). The resulting solution was cooled to -70°C and n-BuLi (2.5 M in hexane, 1.5 eq.) was added slowly. After addition was complete, a solution of 3-fluoroanisole (1.0 eq) in THF (6 vol) was slowly added and kept at -70°C for 5 minutes. B(EtO) ₃ (2.0 eq) was added slowly and kept at -70°C for 10 min. The reaction mixture was quenched with 2 N. NS1. The quenched reaction mixture was extracted with MTBE (3x4 vol). The combined organic phases were concentrated to 1.5-3 total volumes. Heptane was added dropwise (7-9 vol.) and the mixture was cooled to 0-10°C and stirred for 3 hours. The mixture was filtered and washed with heptane (1.5 vol.). The solid was dried under nitrogen at <30°C to give (2-fluoro-6-methoxyphenyl)boronic acid.

Reaction scheme and loading table for step A2.

(2-Fluoro-6-methoxyphenyl)boronic (2-Fluoro-6-hydroxyphenyl)boronic acid acid

Material No. CAS MW (g/mol) Equivalents/volumes mole Weight (G) Volume (l) (2-Fluoro-6- methoxyphenyl)boron 78495 -63-3 169.95 1.0 0.118 20 n. d. alkaline acid Boron tribromide 10294 -33-4 250.52 1.5 0.177 44.2 0.01 7 Dichloromethane 75- 09-2 84.93 4 rev. n. d. n. d. 0.08 0 Water 7732- 18-5 18.02 13 rev. n. d. n. d. 0.26 Methyl tert-butyl ether 1634- 04-4 88.15 13 rev. n. d. n. d. 0.26 Heptane 142- 82-5 100.20 10 rev. n. d. n. d. 0.20

Reactor A was charged with dichloromethane (4 vol.) and 2-fluoro-6-methoxy-4-methylphenylboronic acid (1 eq.). The reaction mixture was cooled to -30°C, and 1.5 BBg ₃ (1.5 eq.) was added dropwise. After addition was complete, the mixture was heated to 25 °C and stirred for 2 hours. The reaction mixture was quenched in ice-cold (0-5 °C) water (10 vol.). MTBE (10 vol) was added and the mixture was heated to 25°C and stirred

-41 045678 for 1-2 hours or until all solids are dissolved. The aqueous phase was separated and extracted with MTBE (3 vol). The combined organic extracts were washed with water (3 vol) and then concentrated to 1 total volume. Heptane (10 vol) was added to the mixture and stirred for 2 hours. The resulting product was isolated by filtration and dried at <30°C to give (2fluoro-6-hydroxyphenyl)boronic acid.

Reaction scheme and loading table for step A3.

But KF ^Η °)=\citric acid _p /\ (H0) ₂ v-^^ --------► ^KF ' ^B- W F acetonitrile, water

Boronic acid Boronate

Material CAS No. MW (g/mol) Equivalents/volumes mole Weight (kg) Volume (l) (2-Fluoro-6hydroxyphenyl)boronic acid 1256345 -60-4 155.92 1.0 89.79 14.00 n. d. Citric acid monohydrate 5949- 29-1 210.14 1.64 147.26 30.94 And. d. Acetonitrile 75-05-8 41.05 21 rev. n. d. 220.1 294 Potassium fluoride 7789- 23-3 58.10 4.00 359.16 20.87 And. d. USP grade water 7732- 18-5 18.02 2.0 rev. n. d. 28.00 28.00 Celite n. d. n. d. n. d. n. d. 7.00 n. d. 2-Propanol 67-63-0 60.10 25 rev. n. d. 275 350

Stage A3.

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 minutes. A separate reactor (reactor B) was charged with (2-fluoro-6hydroxyphenyl)boronic acid (14.00 kg, 89.79 mol), followed by acetonitrile (206.1 kg) and citric acid (30.94 kg, 147.26 mol) at 25 C. The contents of reactor A were added to reactor B at 25°C and stirred at this temperature for 10 hours. The reaction mixture was filtered through a pad of celite (7.0 kg) and washed with acetonitrile (42 kg). The filtrate was combined with isopropanol (56 kg) and then distilled in vacuo at <35°C, replacing the distilled volume in the reactor with isopropanol, and the procedure was repeated as necessary until the solvent change from acetonitrile to isopropanol was completed. The slurry was cooled to 15°C and kept for 1 hour before being filtered and washed with 28 kg of isopropanol. The filter cake was dried using vacuum and nitrogen and packaged to obtain Compound A3.

Separation of the M-dione representing compound 5.

Chromatographic separation of the M-dione intermediate.

Numerous chiral chromatographic techniques and methods were used to isolate Mdione from compound 4. The techniques and stationary phases are well known in the art and are described in Table 1. 1.

- 42 045678

Table 1

Methodology Stationary phase Mobile phase Output ^l SFC Chiralpak® AD 40% methanol/60% CO2 * -95% HPLC Chiralpak® AD 90/10/0.1 ethanol/methanol/triethylamine -94% HPLC Chiralpak® IG 60/40/0.1 ethanol/methanol/ triethylamine -92% Pseudo-moving layer (SMB) Chiralpak® IC Acetonitrile - 96%

^L Yield is defined as the % of available M-dione that was recovered at a required purity of >98%.

* This separation was carried out several times. For each batch of material, the mobile phase could be slightly modified to accommodate batch variations. Additional mobile phases used for purification included:

1) 25/75 methanol/CO ₂ ,

2) 30/70 methanol/CO ₂ , and

3) 50/50 methanol/ _CO2 .

SFC, HPLC and SMB techniques are well known in the art, and Chiralpak® stationary phases are commercially available from commercial sources such as Fisher Scientific and Daicel Corporation.

However, it is necessary to develop a more efficient method for isolating the M-dione (compound 5)

Classic division.

The present invention is directed to the development of a useful classical separation method for the M/P-dione racemate (compound 4).

A total of 100 cocrystal screening experiments were performed and three potential dione cocrystals were identified. Based on the highest M/Pdione area ratio in the residual solid and the lowest area ratio in the supernatant, (+)-2,3-dibenzoyl-O-tartaric acid (DBTA) was selected as the chiral reagent for the separation.

Based on the results of 100 cocrystal screening experiments and 20 additional solvent screening procedures, 2-MeTHP/n-heptane was found to provide better separation performance than other solvent systems. Based on the results regarding the solubility of the M-dione cocrystal and the P-dione cocrystal in various ratios of 2-MeTHF and n-heptane, 2-MeTHF/n-heptane (1.4:1, v/v) was selected as the optimal solvent composition for the separation. about.).

In order to identify any possible conversion of the dione or M/P-dione racemate form during the chiral resolution crystallization process, the solubility in 2-MeTHP/n-heptane (1.4:1, v/v) of the M-dione cocrystal, P-dione cocrystal, mixture of M+P-dione cocrystals (1:1, w/w), dione racemate, and DBTA were determined at different temperatures. No shape change was observed for the M-dione cocrystal and the P-dione cocrystal at different temperatures for 7 days. However, the type C dione racemate was obtained after stirring a mixture of M+P-dione cocrystals (1:1, w/w) at different temperatures for 7 days. The presence of type D dione racemate (20 and 30°C) or type C dione racemate (40, 50, 60 and 65°C) was observed after stirring the dione racemate at corresponding temperatures for 7 days. A solubility of -100 mg/mL was observed at all temperatures for DBTA.

To further optimize the separation method, a ternary phase diagram was generated for the M/P-dione cocrystal based on the equilibrium solubility results, and the eutectic point was not obtained, likely due to the fact that the type C racemate could crystallize in the presence of both the M-dione cocrystal and and R-dion. Another ternary phase diagram for the M/P dione was constructed based on the equilibrium solubility results, and the eutectic point was not obtained, probably because the racemate of the type C or type D dione could crystallize in the presence of both the M dione and P -dione.

Thus, a chiral reagent (DBTA) and a solvent system ((2MeTHF/n-heptane (1.4:1, v/v))) have been identified for the resolution of the dione racemate. A small-scale crystallization method using a resolution reagent and solvent system can provide 39% yield and 99% purity for M-dione.In addition, polymorphism of the dione racemate was observed and investigated during screening experiments.

2. Screening experiment.

2.1. Screening of cocrystals.

-43045678

A total of 100 cocrystal screening experiments were conducted using 20 acids and 5 solvent systems (results are summarized in Table 2-1). In general, the dione racemate and acid in a 1:1 molar ratio were mixed and stirred at room temperature for 3 days before isolation for XRPD. XRPD results identified three potential acids that could form cocrystals with the dione racemate, including (1S)-(-)-camphanic acid (Figure 2-2), (+)-2,3-dibenzoyl-D-tartaric acid ( Fig. 2-3) and D-(+)-malic acid (Fig. 2-4). Based on the XRPD results, four new free base crystal forms were also obtained, which were assigned to the dione racemate of types B-E.

As shown in table. 2-2, the supernatant and residual solid for three potential cocrystals were further tested by HPLC. The M-dione/P-dione ratio was measured and summarized in Table. 2-2. As a result, the cocrystal with DBTA exhibited an M-dione/P-dione area ratio of 0.11 for the supernatant and 4.4 for the residual solid, indicating that M-dione and P-dione exhibited a high degree of separation after cocrystal formation with DBTA. Thus, DBTA was chosen as a chiral reagent to further optimize the separation.

Table 2-1

Summary of data regarding cocrystal screening experiments

Solvent Acid Acetone n ₂ o/ac N (1:1, v/v) EtOAc 2MeTHE/nheptane (1:1, MTBE/nheptane (1:1, v/v) —— ....... v/v) Absent Type B ⁺ Type C ⁺ Type C ⁺ Type D ⁺ Type E ⁺ L-Aspartic acid Type B Type C Type C+acid Type D Type E (R)-1,4-Benzodioxane-2carboxylic acid Type B Type C TypeC Type D Type E+acid (18)-(-)-Camphanovaya acid Type B Type C Cocrystal type A* Cocrystal type B* Cocrystal type A* (-)-Camphoric acid Type B Type C+acid Type C+acid Type D Type E+acid (+)-2,3-Dibenzoyl-D- wine acid Type B Type C Amorphous Cocrystal l type A ^# Cocrystal l type A ^# D-Glutamic acid Type B TypeC Type C+acid Type D Type E B-(+)-Malic acid Type B TypeC Cocrystal type A ^$ Type D Type E (R)-(-)-Almond acid Type B Type C Type C Type D Type E+acid (-)-Methyloxyacetic acid Type B TypeC TypeC Type D Type E (S)-(+)-a- Methoxyphenylacetic acid Type B Type C Type C Type D Type E U)-(+)-a-Methoxy-atrifluoromethylphenylacetic acid Type B Type C Type C Type D Type E (R)-(-)-5-Okco-2- tetrahydrofurancarbono alkaline acid Type B Type C Type C Type D Type E (R)-(+)-N-(l- Phenylethyl)succinamic I'm acid Type B Type C Type C Type D Type E+acid (8)-(+)-2- Type B Type C TypeC Type D Type E

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Phenylpropionic acid L-Pyroglutamic acid Type B Type C Type C Type D Type E+acid O-(-)-Quinic acid Type B+sour Type C Type C+acid Type D+acid Type E+acid L-(+)-Tartaric acid Type B Type C Type C Type D Type E L-Ascorbic acid Type B+sour TypeC Type C+acid Type D Type E N,N-Bhc[(R)-(-)-1phenylethyl]phthalamic acid Type B Type C Type C Type D Type E+acid (8)-Phenyluccinic acid Type B TypeC Type C+acid Type D Type E+acid

⁺ Denotes the crystalline form of the free rac-dione (no cocrystals are formed), *: cocrystal with (1S)-(-)-camphanic acid, #: cocrystal with (+)-2,3-dibenzoyl-D-tartaric acid, $: cocrystal with D-(+)-malic acid.

Table 2-2

Summary of HPLC results for three cocrystals

Peak area Sample Supernatant Solid P-dion (area) M- dione (area) M/R R-dion (area) M- dione (area) M/R Cocrystal with (IS)-(-)- camphanic acid type A 9021.4 8274.2 0.9 6418.6 6360.4 1.0 (810465-06-SZ) Cocrystal with (IS)-(-)camphanic acid type B 4673.2 4303.4 0.9 4768.3 4736.9 1.0 (810465-06-D3) Cocrystal with DBTA type A (810465-06-D5) 17673.1 1858.6 0.11 1180.2 5249.8 4.4 Cocrystal with O-(+)-malic 11382.6 10696, 0.9 6443.3 6366.7 1.0 acid type A 5 (810465-06-С7)

2.2. Solvent screening.

To select a suitable solvent for further separation of M-dione and P-dione, HPLC M/P-dione area ratios were obtained for an additional 20 solvent/solvent mixtures. As indicated in table. 2-3, 2-MeTHF showed the highest degree of separation with an M-dione/P-dione area ratio of 0.7 in the supernatant and 4.1 in the residual solid. However, 2-MeTHF/n-heptane (1:1, v/v) showed higher resolution in the cocrystal screening (Table 2-2), and therefore 2-MeTHF/n-heptane was selected for further optimization. The M/P-dione area ratios from HPLC results were obtained for different ratios of 2-MeTHF/n-heptane at different acid/base ratios. The results shown in table. 2-4 demonstrated that a higher acid/FB ratio (2:1 or 1.5:1) in 2-MeTHF/n-heptane (8:1 or 4:1, v/v) is desirable for improving the M/P-dione ratio in isolated solids.

Determination of the solubility of the M-cocrystal and P-cocrystal in various ratios of 2MeTHF/n-heptane was also carried out at 5 and 25°C, the results of which are summarized in table. 2-5. The M-cocrystal was provided by the customer, and the P-cocrystal was prepared through inverse antisolvent and antisolvent (see Section 4.3 for experimental details). The results regarding solubility are given in table. 2-5 demonstrated that a 2-MeTHF/n-heptane volume ratio of 1.5:1 could provide the highest degree of separation at room temperature. Additional separation experiments were carried out by the client, resulting in the ratio

- 45,045,678 volumes of 1.4:1 demonstrated the highest degree of separation. Therefore, a volume ratio of 2-MeTHP/H-heptane of 1.4:1 was selected as the solvent system for separation.

Table 2-3

Solvent screening for the cocrystal of the dione racemate with DBTA (ratio of M________________________dione/P-dione area values)________________________

Solvent Supernatant Solid Will dissolve spruce Supernatant Solid 2-MeTHF 0.7 4.1 DMSO/heptane (1:1, v/v) n. d. n. d. MTBE 0.04 3.5 DMF/heptane (1:1, v/v) 0.9 1.0 MeOH 0.9 1.0 Toluene/n- 0.8 1.0 heptane (1:1, v/v) IPA 1.0 1.0 Acetic acid n. d. n. d. EtOH/nheptane (1:1, v/v) 0.9 1.0 Muravyina I'm acid n. d. n. d. MIBK 0.9 1.0 DCM n. d. n. d. MEK 1.0 1.0 Cumene 0.9 1.0 IP Ac 0.9 1.0 1 -Butanol 0.9 1.0 THF n. d. n. d. n- Propanol 0.9 1.0 NMP n. d. n. d. 1,3Dimethyl-2imidazolide inone 0.9 1.0

N.D.: a clear solution was obtained, but no solid substance was isolated.

Table 2-4

Screening results for acid/base ratio and 2-MeTHP/n-heptane ratio for dione racemate cocrystal with DBTA (M-dione/P-dione area ratio)

\Ratio \kisyaota/osn ovation 2-MeTHNH/nheptane \ (v/v) \ 2:1 1.5:1 1:1.5 1:2 For ma L S For ma L S For ma L S For ma L S 8:1 SA 0.4 13.4 SA 0.4 8.9 SA 0.5 7.0 SA 0.6 2.6 4:1 SA o, s 8.7 SA o, s 6.2 SA 0.4 6.2 SA 0.4 5.7 2:1 SA 0.2 6.0 SA 0.2 5.6 SA 0.2 4.7 SA 0.2 5.2 1:2 SA o, s 1.2 SA 0.1 1.3 SA 0.1 1.4 SA 0.1 1.7 1:4 D.A. 1.4 1.0 SA 0.4 1.0 SA + D.A. 0.1 1.0 SA 0.1 1.0 1:8 SA 0.9 1.0 D.A. 0.5 1.0 D.A. o, s 1.0 D.A. o, s 1.0

-46045678

+ D.A.

L: supernatant, S: solid, CA: type A cocrystal, DA: type A dione racemate.

Table 2-5

Screening of 2-MeTHF/n-heptane ratio for M/P-dione cocrystals

Temperature (°C) 2-MeTShn- heptane (v/v) M-dione cocrystal P-dione cocrystal Solubility (mg/ml) XRPD Soluble (mg/ml) XRPD 5 1:1 11.3 Type A* 38.5 Type A ^# 1.5:1 17.3 Type A* 60.8 Type A ^# 2:1 22.2 Type A* 66.9 Type A ^# 3:1 28.6 Type A* 86.6 Type A ^# 4:1 30.8 Type A* 82.9 Type A ^# 6:1 47.1 Type A* 92.1 n. d. 8:1 55.1 Type A* 90.6 n. d. 25 1:1 13.4 Type A* 52.9 Type A ^# 1.5:1 20.3 Type A* 80.8 Type A ^# 2:1 28.1 Type A* 81.7 Type A ^# 3:1 39.0 Type A* 87.0 n. d. 4:1 43.0 Type A* 86.5 n. d. 6:1 53.2 Type A* 81.9 n. d. 8:1 65.0 Type A* 89.2 n. d.

*: M-cocrystal type A, #: P-cocrystal type A.

2.3. Solubility of dione cocrystal with DBTA, dione racemate and DBTA.

A 7-day equilibrium solubility test of M-dione cocrystal, P-dione cocrystal, M+P-dione cocrystal mixture (1:1, w/w) and dione racemate was carried out at different temperatures (20, 30, 40, 50, 60, 65, 75 and 80°C) in 2-MeTHF/n-heptane (1.4:1, v/v). A color change was observed at 75 and 80°C after 5 days, indicating degradation, so no solubility data was obtained.

When mixing the M-cocrystal and P-cocrystal at different temperatures for 7 days, no change in shape was observed (Fig. 2-5 and Fig. 2-6). The type C dione racemate was prepared after stirring a mixture of M-dione cocrystals and P-dione cocrystals (1:1, w/w) at different temperatures for 7 days (Figures 2-7). Type D dione racemate (20 and 30°C) or type C dione racemate (40, 50, 60 and 65°C) were observed after stirring the dione racemate at various temperatures for 7 days (Fig. 28 and Fig. 2-9) .

A five-day equilibrium solubility test of DBTA was carried out at different temperatures (20, 30, 40, 50, 60 and 65°C) in 2-MeTHF/n-heptane (1.4:1, v/v). Solubility of ~100 mg/mL was observed at all temperatures. No significant difference was observed with temperature changes (Table 2-7).

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Table 2-6

Solubility of dione cocrystal with DBTA, mixture of M/P-dione cocrystals, dione racemate in 2MeTHF/n-heptane (1.4:1, v/v)

Sample ID Material Temperature (°C) Solubility (mg/ml) Crystal form M-dion P-dion 1-01-A1 Cocrystal M-dione 20 13.1 - M-type cocrystal A 1-01-A2 thirty 15.8 - M-type cocrystal A 1-01-AZ 40 18.4 - M-type cocrystal A 1-01-A4 50 17.2 - M-type cocrystal A 1-01-A5 60 34.6 - M-type cocrystal A 1-01-Ab 65 35.4 - M-type cocrystal A 1-01-B1 Rdion cocrystal 20 - 39.4 P-type cocrystal A 1-01-B2 thirty - 56.5 P-type cocrystal A 1-01-VZ 40 - 55.8 P-type cocrystal A 1-01-B4 50 - 79.9 P-type cocrystal A 1-01-B5 60 - 113.9 P-type cocrystal A 1-01-Vb 65 - 110.0 P-type cocrystal A 1-01-S1 Mixture cocrystals of M+P-dione 20 7.3 10.4 Dione type racemate WITH 1-01-S2 thirty 9.2 16.0 Dione type racemate

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WITH 1-01-SZ 40 9.8 12.2 Dione type racemate WITH 1-01-С4 50 12.1 21.9 Dione type racemate WITH 1-01-С5 60 18.7 26.7 Dione type racemate WITH 1-01-С6 65 13.4* 18.0* Dione type racemate WITH 1-01-D1 Dione racemate 20 18.0 15.2 Dione type racemate D 1-01-D2 thirty 20.1 17.1 Dione type racemate D 1-01-D3 40 I.5 9.9 Dione type racemate WITH 1-01-D4 50 14.2 I,8 Dione type racemate WITH 1-01-D5 60 13.7 I,7 Dione type racemate WITH 1-01-D6 65 15.3 13.1 Dione type racemate WITH

Table 2-7

Solubility of DBTA in 2-MeTHF/n-heptane (1.4:1, v/v)

Sample ID Temperature (°C) Solubility (mg/ml) 1-15-A1 20 99.1 1-15-A2 thirty 100.3 1-15-AZ 40 98.9 1-15-A4 50 88.0 1-15-A5 60 105.0 1-15-A6 65 96.1

2.4. Triple phase diagram.

2.4.1. M/P-dione cocrystals.

The M-dione cocrystal and the P-dione cocrystal were weighed, and the corresponding mass values are indicated in the table. 2-8, and stirred in 2-MeTHF/n-heptane (1.4:1, v/v) at room temperature for 72 hours. The ternary phase diagram of the M/P-dione cocrystal was generated based on the 72-hour equilibrium solubility data, and the eutectic point was not obtained (Fig. 210).

Table 2-8

Summary of solubility data for M/P-dione cocrystals

No. Weight of Msocris talla (mg) Weight of Rsocris thallus (mg) De (%) Supernatant Solid [M], mg/m l [P], mg/m l De (%) [M]/[ R] m, mg P, mg De (%) 1 50.7 0 100.0 22.9 0.0 100.0 n. d. 27.8 oh oh 100.0 2 0 99.7 -100.0 0.0 81.2 -100.0 n. d. oh oh 18.5 -100.0 3 21.1 83.4 -59.6 5.1 59.4 -84.2 11.68 0 16.0 24.0 -20.0 4 41 83.5 -34.1 5.9 45.0 -76.9 7,658 35.1 38.5 -4.6 5 84 85 -0.6 I.5 23.7 -34.9 2,070 72.5 61.3 8.4 6 167.7 83.1 33.7 12.2 20.8 -26.2 1.711 155.5 62.3 42.8 7 49.7 50.4 -0.7 17.5 40.8 -40.0 2,331 32.2 9.6 54.1 8 49.7 25 33.1 19.9 21.8 -4.6 1.095 29.8 3.2 80.7 9 49.2 12.2 60.3 21.3 9.6 37.9 0.451 27.9 2.6 82.9

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2.4.2. M/P-dione.

M-dione and P-dione were weighed, and the corresponding mass values are indicated in the table. 2-9, and stirred in 2-MeTHF/n-heptane (1.4:1, v/v) at room temperature for 5 days. The ternary phase diagram was constructed from 5-day equilibrium solubility data in 1.0 ml 2-MeTHF/n-heptαnα (1.4:1, v/v) at room temperature. Type A M-dione, Type A P-dione, Type C and Type D dione racemate were observed in the residual solid of the solubility testing samples. No eutectic points were obtained on the phase diagram (Fig. 2-11).

Table 2-9

Generalized data regarding the ternary phase diagram for the M/P dione

No. Weight Mdion a (mg) Weight R- DIO per (mg) De (%) Supernatant solid XRP D [M m mg/ ml [P], mg/ml De (%) [M]/[R] M, mg P, mg De (%) 1 99.3 0 100.0 80, 1 0 100.0 n. d. 19.2 oh oh 100.0 A 2 0 100, -100.0 0 77.1 -100.0 N. d. oh oh 23.5 -100.0 A 6 3 20.2 119, 3 -71.0 7.7 50.3 -73.6 0.2 12.5 69.0 -69.2 A+R D 4 89.8 90.3 -o, s 16, 2 17.5 -3.9 0.9 73.6 72.8 0.6 R.D. 5 121.1 40.7 49.7 53, 0 1.6 94.2 33.4 68.1 39.1 27.0 A+R WITH 6 90.2 19.5 64.4 52, 6 1.4 94.8 37.6 37.6 18.1 35.0 A+R WITH 7 41.6 39.9 2.1 9.8 9.0 4.4 1.1 31.8 30.9 1.4 R.C. 8 39.2 120, 3 -50.8 7.6 48.1 -72.8 0.2 31.6 72.2 -39.1 A+R D 9 119.1 20.5 70.6 54, 1 1.6 94.4 34.5 65.0 18.9 54.9 A+R WITH

RC: type C dione racemate; RD: type D dione racemate; A: M- or P-dione type A (powder X-ray diffraction patterns for M-dione type A and P-dione type A were the same and did not differ).

Characteristics of the solid state of crystalline forms.

A total of five crystalline forms of the dione racemate and two cocrystal forms were obtained. All these forms were characterized by XRPD, TGA, DSC, PLM and 1H NMR and summarized in Table. 2-10. Solid state characterization data indicated that type A and type D dione racemates were identified as 2-MeTHF solvates, type B as acetone solvate, type C as anhydrate, and type E as MTBE solvate.

It was found that both the type A M-dione cocrystal and the type A P-dione cocrystal were present as solvates with 2-MeTHF. All data regarding the characteristics are presented in Fig. 3-1 - fig. 3-22.

Table 2-10

Summary of data regarding crystal forms

ID sample Crystal form Endotherm (peak, °C) TGA (wt.%) W NMR (wt.%) 5-05-A Type A dione racemate 110.2, 248.6, 213.4* 2.5 (150 °C) 2.4 (2-MeTHF) 1-10-A1 Type B dione racemate 113.4, 126.0, 250.9 9.8 (150 °C) 6.2 (acetone) 1-01-D5 Type C dione racemate 251.9 3.0 (150 °C) ND ^& 1-01-D1 Type D dione racemate 120.5, 253.3 15.4 (150 °C) 12.0 (2-MeTHF)

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1-10-A4 Type E dione racemate 151.6, 158.6, 248.7 14.7(160°C) 7.5 (MTBE) 5-17-A Type A M-dione cocrystal 109.6, 119.2 6.6 (125°C) 10.6 (2-MeTHF) 5-16-A Type A P-dione cocrystal 88.3, 112.3, 132.8 9.2 (140°C) 10.6 (2-MeTHF)

*: exothermic peak; &: not detected.

3.1.1. Competitive suspension of dione racemate forms.

Types B-E dione racemate was successfully re-prepared by suspending type A dione 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) at room temperature, respectively.

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

All free base forms were converted to the type C dione racemate by competitive suspension in 2-MeHTF/n-heptane (1.4:1, v/v) at target temperatures, suggesting that the type C dione racemate is the most thermodynamically stable form in 2-MeHTF/Hheptane (1.4:1, v/v) in the range from 20 to 65°C.

Table 2-11

Results regarding competitive suspension

Original form ID experiment Solvent Temperature (°C) Solid form Dione racemate types A-E 1-16-B1 2-MeTHE/n- heptane (1.4:1, v/v) 20 Type C dione racemate 1-16-B2 thirty Type C dione racemate 1-16-VZ 40 Type C dione racemate 1-16-B4 50 Type C dione racemate 1-16-B5 60 Type C dione racemate 1-16-B6 65 Type C dione racemate

3.2. Preparation of P-dione cocrystal.

3.2.1. Small scale.

g P-dione and 1 g DBTA were dissolved in 18 ml 2-MeTHF at 65°C to obtain an almost clear solution. 18 ml of heptane was added to this solution after 1 hour. The solution was cooled to 20°C over 4 hours and kept overnight. The solution was evaporated by blowing air at room temperature for approximately 1 hour to give a yellowish oily paste. Another 54 ml of heptane was added to the mixture with stirring for 2 hours. The suspension was filtered. The solid sample was designated 810465-16-A.

3.2.2. Large scale.

g P-dione and 5 g DBTA were dissolved in 100 ml 2-MeTHF at 65°C. The solution was filtered using a PTFE filter with a pore size of 0.45 μm to obtain a clear solution. The clear solution was added dropwise to a 400 mL heptane suspension containing ~1 g of seed (810465-16-A) obtained from the first run. The suspension was kept with stirring at room temperature for 5 hours before isolation. Approximately 10 g of P-dione cocrystal (810465-20-A) was obtained in 66% yield.

4. Devices and methods.

4.1. XRPD.

PANalytical X-ray powder diffractometers in reflection mode were used for XRPD analysis. The applied XRPD parameters are given in table. 4-1.

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Table 4-1

XRPD Test Options

Options PANalytical PANalytical PANalytical Model Empyrean X'Pert ³ X'Pert ³ The length of the waves! x-ray 1uchey Si, ka, Kal (A): 1.540598, Ka2 (A): 1.544426, intensity ratio Ka2/Kal: 0.50 Si, ka, Kal (A): 1.540598, Ka2 (A): 1.544426, intensity ratio Ka2/Kal: 0.50 Si, ka, Kal (A): 1.540598, Ka2 (A): 1.544426, intensity ratio Ka2/Kal: 0.50 X-ray tube parameters 45 kV, 40 mA 45 kV, 40 mA 45 kV, 40 mA Del divergences Installed automatically 1/8° Fixed at 1/8° ’mode zhanization Continuous Continuous Continuous Range Options zhanization ;°2ТН) Time step zhanirovaniya (s) 3°-40° PANalytical 17.8 3°-40° PANalytical 46.7 3°-40° PANalytical 18.9 ’step size (°2TN 0.0167 0.0263 0.0131 Test time 5 min 30 s 5 min 04 s 4 min 15s

4.2. TGA and DSC.

Data regarding TGA were collected using TA discovery 550, Q500 and Q5000 TGA from TA Instruments. DSC was performed using Q500, Q5000 and Discovery 2500 DSC from TA Instruments. Details regarding the parameters used are given in Table. 4-2.

Table 4-2

Test procedure parameters via TGA and DSC Options TGA DSC Way Linear change Linear change Sample cuvette Aluminum, open Aluminum, corrugated Temperature Room temperature 35°C 25°С - 300°С Heating rate 10°C/min 10°C/min Purge gas n ₂ n ₂ 4.3. HPLC.

An Agilent 1100/1260 HPLC instrument was used for solubility testing, with details regarding the methods given in Table. 4-3.

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Table 4-3

HPLC Solubility Testing Method

HPLC Agilent 1100 with DAD detector Column CHIRALPAK IC-3, 4.6x100 mm, 3 µm Mobile phase A: n-heptane B: MeOH/EtOH (1:1, v/v) Isocratic elution A:B=75:25, 60:40 Duration of analysis 10.0 min Break time 0.0 min Flow rate 1.0 ml/min Injected volume 5 µl Detector wavelength UV at 215 nm

Column temperature 40°C Sampler temperature Room temperature Diluent EtOH

Table 4-4

Dissolution Test Method (DBTA) by HPLC

HPLC Agilent 1260 with DAD detector Column Agilent ZORBAX 300SB-C3, 150 x 4.6 mm, 3.5 µm Mobile phase A: 0.05% TFA in _H2O B: 0.05% TFA in ACN Isostatic elution A:B=65:35 Duration of analysis 5.0 min Break time 0.0 min Flow rate 0.6 ml/min Injected volume 5 µl Detector wavelength UV at 215 nm Column temperature 40°C Sampler temperature Room temperature Diluent EtOH

4.4. 1H NMR.

1H NMR spectrum was obtained using a Bruker 400M NMR spectrometer using DMSO-d ₆ as solvent.

4.5. PLM.

Polarized light microscopic images were obtained on a Nikon DS-Fi2 upright microscope at room temperature.

Additional screening of compound 5 with 1,3-diphenyl-3-oxopropanesulfonic acid 11b

Due to the low basicity of the pyridine moiety in compound 5 and limited hits in terms of formation of crystalline salts using a standard screening kit, it was selected to screen racemic compound 4 with 1,3-diphenyl-3oxopropanesulfonic acid 11b on a 0.06 mmol scale.__________________________

O NO.j 1.0 S o S/Ae O BUT,.and, O S about o BUT„d,O S O 11-RAC 11a 11b (+)-enantiomer (-)-enantiomer

Resolving power of racemic compound 5 on a gram scale. A 250 mL round bottom flask was charged with 2.0 g of racemic compound 4 (5.7 mmol, 1.0 eq) in 200 mL EtOH:AcOH (90:10 v/v). After dissolving the material, 832 mg of sulfonic acid 11b (2.9

- 53 045678 mmol, 0.5 eq.). The clear solution was left to stir for 15 hours at a stirring speed of 800 rpm. A white precipitate formed, which was isolated from the original solution. The isolated salt was suspended in CH2Cl2, which was treated with concentrated aqueous NaHCO3 using a separatory funnel. The organic layer was isolated and the main aqueous layer was extracted with CH2Cl2 (2x). The organic layers were combined and dried over Na2SO3. Evaporation of the solvent provided 415 mg (M)-5 (ee 96%) (see Fig. 6-1).

The clear initial solution was evaporated to dryness. The yellow oily material was dissolved in CH ₂ Cl ₂ and treated with concentrated aqueous NaHCO ₃ using a separatory funnel. The organic layer was isolated and the main aqueous layer was extracted with CH ₂ Cl ₂ (2x). The organic layers were combined and dried over Na2SO3. Evaporation of the solvent provided 1579 mg of 5 (23% ee for the (P)-atropoisomer; Figure 6-2).

Screening of polymorphs of M-dione cocrystal with DBTA.

5. Characteristics of the crystalline forms of the cocrystal of M-dione with DBTA.

Polymorph screening experiments for M-dione were performed under 100 conditions using slurry conversion, slow evaporation, slow cooling, antisolvent addition, vapor diffusion, temperature cycling, and wet grinding methods. As a result of screening, a total of 17 crystalline forms (types A-Q) were obtained. The relationship between the shapes is shown in Fig. 4-1. Detailed characteristic data are presented in table. 5-1, and overlays of X-ray powder diffraction patterns are shown in FIG. 5-1. The solid state characterization results indicated that type G was a hydrate, whereas types were solvates.

5.1. Devices and methods.

5.1.1. XRPD.

XRPD was performed using Panalytical X'Pert ³ Powder XRPD on a Si-based zero background holder. Position 29 was calibrated against a reference standard Si-based disk from Panalytical. The applied parameters are listed in table. 5-a.

Table 5-a ____________XRPD Test Parameters____________

Options Reflection mode X-ray wavelength Si, ka, Καί (A): 1.540598, Ka2 (A): 1.544426, intensity ratio Ka2/Ka1: 0.50 X-ray tube parameters 45 kV, 40 mA Divergence slot Fixed at 1/8° Scan mode Continuous Scan range (°2TH) 3-40 Scan step time [s] 18.87 Step size (°2TN) 0.0131 Test time 4 min 15 s

5.1.2. TGA/DSC.

TGA data were collected using a TA Discovery 550 TGA from TA Instrument. DSC was performed using a TA Q2000 DSC from TA Instrument. DSC calibration was performed using an indium reference standard and TGA calibration was performed using a nickel reference standard. Details regarding the parameters used are given in Table. 5-b.

Table 5-b

Test procedure parameters via TGA and DSC

Options TGA DSC Way Linear change Linear change Sample cuvette Platinum, open Aluminum, corrugated Temperature Room temperature - required temperature Heating rate 10°C/min Purge gas ν ₂

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5.2. Screening of polymorphs.

Type A (3-05-A) solubility was assessed at room temperature. Approximately 2 mg of solid was added to a 3 ml glass vial. Then the solvents from the table were gradually added to the vials. 5-s (50/50/200/700 µl) until solids dissolve or total volume reaches 2 ml. The results summarized in table. 5-c was used as a guide for solvent selection during polymorph screening.

Polymorph screening experiments were carried out using various crystallization or solidification methods. The methods used and the crystal types identified are summarized in Table. 5-s.

Table 5-c

Approximate solubility of the starting material (6010013-05-A) at room temperature

Solvent Solubility (mg/ml) Solvent Solubility (mg/ml) MTBE S<3.1 Acetone S>52.0 H ₂ O 2.4<S<8.0 DMF S>40.0 n-Geptane 3.0<S<10.0 Anisole S>40.0 Toluene 2.7<S<9.0 Acetic acid S>40.0 Hexanes 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 CHC13 S>50.0 1,4-Dioxane 25.0<S<50.0 EtOAc S>72.0 n-ViON 23.0<S<46.0 DMSO S>72.0 MIBK 38.0<S<76.0 MeOH S>78.0 ViOAc S>28.0 EtOH S>68.0 IP Ac S>32.0 — —

Table 5-d

Summary of data regarding polymorph screening experiments

Way Number experiments Crystal type Suspension at room temperature temperature/5°С 37 Types A-G, Type J, Type N, Type N Slow evaporation 16 Type A, Type C, Type D, Type J, Type K, Type L, Type N, Type O Slow cooling 9 Type C, Type J, Type L, Type O Adding Antisolvent 9 Type A, Type C, Type H and Type I Diffusion at the liquid-vapor interface 5 Type L, Type M, Type Q Diffusion at the solid-vapor interface 6 Types A and M Cyclic change temperature 7 Type A, Type G, Type O Wet grinding 10 Type A Total 99 Types A-Q

5.2.1. Suspend at room temperature.

Experiments with suspensions were carried out at room temperature in various solvent systems. Approximately 20 mg of type A (3-05-A) was suspended in 0.2 ml of solvent in a 3 ml glass vial. After stirring the suspension using a magnetic stirrer for 13 days at room temperature, the remaining solids were isolated for analysis by XRPD. The results summarized in table. 5th, indicate that types A-D and type J were received.

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Table 5-e

Obobg Attribute valuable data. Experiment ID flax experiments with suspensions Solvent (vol:vol) o at room temperature Solid form ) ature 3-07-A1 MTBE Type B 3-07-A2 n ₂ o Type A 3-07-AZ n-Geptane Low crystallinity 3-07-A4 Toluene Low crystallinity 3-07-A5 Hexanes Type A 3-07-A6* IPA Type A 3-07-A7* 2-MeTHF Type B 3-07-A8* 1,4-Dioxane Type J 3-07-A9 n-ViON Low crystallinity 3-07-A10* MIBK Type A 3-07-A11 ViOAc Type C 3-07-A12 IP Ac Type D 3-07-A13* Acetone Amorphous 3-07-A14* DMF Type P 3-07-A15 Anisole Type E 3-07-A16 THF/n-heptane (1:9) Low crystallinity 3-07-A17 2-MeTHP/n-heptane (1:9) Type A 3-07-A18 1PA/H2O(1:9) Type A 3-07-A19 1RAc/H2O (1:9) Type F 3-07-A20 n-ViOH/H2O (1:9) Type A 3-07-A21 n-ViON/MTBE (1:9) Type A 3-07-A22 SNS13/MTBE (1:9) Type A 3-07-A23* MeOH/H2O (937:63, aw=0.2) Amorphous 3-07-A24* MeOH/H2O (844:156, aw=0.4) Type N 3-07-A25* MeOH/H2O (693:304, aw=0.6) Type G 3-07-A26 MeOH/H2O (569:431, aw=0.8) Type G

*: solid obtained by slow evaporation at room temperature.

5.2.2. Slow evaporation.

Slow evaporation experiments were carried out under 16 conditions. Briefly, 20 mg of type 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 filtrates were used in the following steps. Visually clear solutions were covered with Parafilm® with 5-10 small diameter holes and evaporated at room temperature. Solids were isolated for analysis by XRPD. The results summarized in table. 5-f demonstrated that type A, type C, type D, type J, type K, type L, type N, type O were obtained.

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Table 5-f

Summary of data regarding slow evaporation experiments

5.2.3. Slow cooling.

Slow cooling experiments were carried out in 9 solvent systems. Approximately 20 mg of type 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, equilibrated for two hours and filtered using a PTFE membrane (0.20 μm pore size). The filtrates were slowly cooled to 5°C at a rate of 0.1°C/min. The results summarized in table. 5-g indicate that type C, type G, type J, type L and type O were observed.

Table 5-g

Summary of data regarding slow cooling experiments

*: solids obtained by evaporation at room temperature.

5.2.4. Adding antisolvent.

A total of 9 experiments involving the addition of antisolvent were performed. Approximately 20 mg of starting material (3-05-A) was dissolved in 0.2-1.4 ml of solvent to obtain a clear solution. The solution was stirred using a magnetic stirrer, followed by stepwise addition of 0.2 ml of antisolvent until a precipitate appeared or a total volume of antisolvent reached 15.0 ml. The resulting precipitate was isolated for XRPD analysis. The results shown in table. 5-h showed that type A, type C, type H and type I were obtained.

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Table 5-h

Summary of data regarding antisolvent addition experiments

Ш experiment Solvent Anti-solvent Solid form 3-10-A1* H2O Acetone Type Η 3-10-A2 THF Amorphous W-S-AZ* DMSO Type I 3-10-A4* MTBE EtOH Type A 3-10-A5* SNA13 Type A 3-10-A6 EtOAc Type A 3-10-A7* n-Geptane Acetone Type C 3-10-A8* 2-MeTHF Type A 3-10-A9* IPAc TypeC

*: solids obtained by evaporation at room temperature.

5.2.5. Diffusion at the liquid-vapor interface.

Five experiments on diffusion at the liquid-vapor interface were carried out. Approximately 20 mg of starting material (3-05-A) was dissolved in an appropriate solvent to obtain a clear solution in a 3 ml vial. This solution was then placed in a 20 ml vial with 3 ml of volatile solvents. The 20 ml bottle was hermetically sealed with a lid and kept at room temperature, providing sufficient time for organic vapors to interact with the solution. Precipitates were isolated for XRPD analysis. The results summarized in table. 5-i showed that type L, type M and type Q were obtained.

Table 5-i

Summary of data regarding diffusion experiments at the liquid-vapor interface

Experiment ID Solvent Anti-solvent Solid form 3-11-A1 mshk n-Geptane Type Q 3-11-A2 EtOAc IPA Type M Z-I-AZ THF MTBE Type L 3-11-A4 2-MeTHF n-Geptane Type L 3-11-A5* DMF Toluene Gel

*: Solids were obtained by evaporation at room temperature.

5.2.6. Diffusion at the solid-vapor interface.

Solid-vapor interface diffusion experiments were performed using 6 different solvents. Approximately 10 mg of starting material (3-05-A) was weighed into a 3 ml vial, which was placed in a 20 ml vial with 2 ml of volatile solvent. The 20 ml vial was hermetically sealed with a lid and kept at room temperature for 7 days, allowing the solvent vapor to interact with the sample. Solids were tested by XRPD and the results are summarized in Table. 5-j demonstrated that type A and type M formation occurred.

Table 5-i

Summary of data regarding diffusion experiments at the solid-vapor interface

Experiment ID Solvent Solid form 3-12-A1 EtOH Type M 3-12-A2 MTBE Type A 3-12-AZ H2O Type A 3-12-A4 Acetone Amorphous 3-12-A5 2-MeTHF Type A 3-12-A6 IPAc Type A

5.2.7. Cyclic temperature change.

Temperature cycling experiments were carried out in 7 solvent systems. Approximately 20 mg of starting material (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, equilibrated for one hour and filtered using a PTFE membrane (0.20 μm pore size). The filtrates were slowly cooled to 5°C at a rate of 0.2°C/min and then heated to 50°C at a rate

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1°C/min. The cycle was repeated one more time and then cooling was provided to 5°C at a rate of 0.2°C/min. Samples were stored at 5°C before solids were isolated and analyzed using XRPD. The results summarized in table. 5-g indicate that type A, type G and type O were observed.

Table 5-k

Summary of Temperature Cycling Experiments

*: solids obtained by evaporation at room temperature.

5.2.8. Suspension at 5°C.

Experiments with suspensions were carried out at a temperature of 5°C in various solvent systems. Approximately 20 mg of starting material (3-05-A) was suspended in 0.2 ml of solvent in a 3 ml glass vial. After stirring the suspension using a magnetic stirrer for 7 days at 5°C, the remaining solids were isolated for analysis by XRPD. The results summarized in table. 5-1 indicate that type A, type C - type E and type J were obtained.

Table 5-1

Summary of data regarding suspension experiments at 5°C

Experiment ID Solvent (vol:vol) Solid form 3-14-A1* ViOAc TypeC 3-14-A2* IP Ac Type D 3-14-AZ* 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* SNA13 Low crystallinity 3-14-A9* EtOAc Low crystallinity 3-14-A10* MeOH TypeC 3-14-A11* THF Type J

*: solids obtained by evaporation at room temperature.

5.2.9. Wet grinding.

Wet grinding experiments were carried out under five conditions. Briefly, 10 mg of type A (3-05A) was placed in a mortar and ground in ∼20 μL of solvent for 5 min. Solids were isolated for analysis by XRPD. The results summarized in table. 5-m indicate that type A was received.

- 59 045678

Table 5-m

Summary of data regarding wet grinding experiments

Experiment ID Solvent (vol:vol) Solid form 3-15-A1 MTBE Amorphous 3-15-A2 H ₂ O Amorphous 3-15-AZ n-Geptane Amorphous 3-15-A4 Toluene Amorphous 3-15-A5 Hexanes Amorphous 3-15-A6 IPA Amorphous 3-15-A7 2-MeTHF Type A (low crystallinity) 3-15-A8 1,4-Dioxane Type A 3-15-A9 n-ViON Amorphous 3-15-A10 MIBK Amorphous

Table 5-1

Characterization of crystal forms of M-dione cocrystal with DBTA

Crystal Form (Batch No.) Conditions receiving Weight loss during TGA (%) Endotherm, according to DSC (peak, °C) Form ID Type A (3-05-A) Traditional separation from DBTA (2-MeTHF) 7.31 to 125°С 109.4 120.0 Solvate with 2- MeTHF Type B (3-07-A1) Suspension at room temperature (MTBE) 7.21 to 125°C 115.7 Solvate with MTBE TypeS (3-08-A5) Slow evaporation (EtOAc) 7.99 to 125°С 92.7 116.4 Solvate with EtOAc Type D (3-07-A12) Suspension at room temperature (IP Ac) 7.51 to 130°C 75.4, 110.5, 148.0, 116.6, 265.9(exotherm) Solvate with IP Ac Type E (3-07-A15) Suspension at room temperature (anisole) 8.63 to 125°С 103.8 119.0 Solvate with anisole Type F (3-07-A19) Suspension at room temperature in 1RAc/H ₂ O (vol:vol 1:9) 6.2 to 130°C 86.5, 107.9 Solvate with IP Ac Type G (3-07-A26) Suspension at room temperature (MeOH/H ₂ O, aw=0.8) 6.44 to 100°C 86.0 127.2 133.1 Hydrate

-60045678

TypeN (3-10-A1) Anti-solvent (acetone/H ₂ O) 3.58 to 130°C 107.6 Acetone solvate Type I (W-S-AZ) Anti-solvent (DMSO/H ₂ O) 7.26 to 150 °C. 128.9 Solvate with DMSO Type J (3-08-A2) Slow evaporation (THF) 7.27% at 125°C 115.7 THF solvate Type K (3-08-A14) Slow evaporation (acetone) 5.71 to 150°C 96.7, 119.8 147.4 157.4°C (exotherm) Acetone solvate Type L (3-11-A4) Diffusion to liquid-vapor boundary (2-MeTNT/nheptane) 7.94 to 130 °C 126.2 Solvate with 2- MeTHF Type M (3-11-A2) Diffusion to liquid-vapor interface (EtOAc/IPA) 3.73 to 150°C 122.6 Solvate with IPA ΤηπΝ (3-08-A8) Slow evaporation (EtOH) 4.08 to 150°C 85.4, 126.5 150.9 Solvate with EtOH Type O (3-08-A11) Slow evaporation (MIBK) 2.03 to 130°С 106.2, 151.2 Solvate with MIBK Type P (3-07-A14) Slow evaporation (DMF) 8.33 to 130°C 89.9 Solvate with DMF Type Q (3-11-A1) Diffusion to liquid-vapor boundary (MSH/n-heptane) 6.00 to 120°С 92.9, 148.9’ 170.0 Solvate with MIBK

5.3. Type A.

Type A (3-05-A) was provided by the client. The XRPD result shown in FIG. 5-4, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-5, a 7.3% weight loss up to 125°C and two endotherms at 109.4 and 120.0°C (peak) were observed. As shown in FIG. 5-6, the presence of 2MeTHF was detected in the spectrum of ^! H NMR. Based on the results, type A was considered to be a 2-MeTHF solvate.

5.4. Type B.

Sample type B (3-07-A1) was obtained from a suspension of type A in MTBE at room temperature. The X-ray powder diffraction pattern shown in FIG. 5-7, indicated a crystalline form. As shown by the TGA and DSC data in FIG. 5-10, a 7.2% weight loss up to 125°C and an endotherm at 115.7°C (peak) were observed. As shown in FIG. 5-9, the presence of MTBE was detected in the spectrum ^! H NMR. Based on the results, type B was likely a solvate of MTBE.

5.5. Type C.

Sample type C (3-08-A5) was prepared by slow evaporation in EtOAc at room temperature. The X-ray powder diffraction pattern shown in FIG. 5-10, indicated a crystalline form. The TGA and DSC data shown in FIG. 5-11 showed an 8.0% weight loss up to 125°C and two endotherms at 92.7°C and 116.4°C (peak). As shown in FIG. 5-12, the presence of EtOAc was detected

-61 045678 is present in the 1H NMR spectrum. Based on the results, type C was likely a solvate of EtOAc.

5.6. Type D

Sample type D (3-07-A12) was prepared from a suspension of type A in IPAc at room temperature. The XRPD result shown in FIG. 5-13, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-14, a 7.5% weight loss up to 130°C and endotherms at 75.4°C, 110.5°C, 148.0°C and 116.6°C (peak) were observed, as well as an ectotherm at 265.9°C. As shown in FIG. 5-15, the presence of IPAc was detected in the 1H NMR spectrum. Based on the results, type D was likely a solvate of IPAc.

5.7. Type E.

Sample type E (3-07-A15) was prepared from a suspension of type A in anisole at room temperature. The XRPD result shown in FIG. 5-16, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-17, a weight loss of 8.6% up to 125°C and two endotherms were observed at 103.8°C and 119.0°C (peak). As shown in FIG. 5-18, the presence of IPAc was detected in the 1H NMR spectrum. Based on the results, type E was likely an anisole solvate.

5.8. Type F

Sample type F (3-07-A19) was prepared from a suspension of type A in IPAcH ₂ O (v:v 1:9) at room temperature. The XRPD result shown in FIG. 5-19, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-20, a 6.2% weight loss up to 130°C and two endotherms were observed at 86.5°C and 107.9°C (peak). As shown in FIG. 5-18, the presence of IPAc was detected in the 1H NMR spectrum. Based on the results, type F was likely a solvate of IPAc.

5.9. Type G

Sample type G (3-07-A26) was prepared from a suspension of type A in MeOH ₂ O (aw=0.8) at room temperature. The XRPD result shown in FIG. 5-22, indicated a crystalline state. As shown by the TGA and DSC data in FIG. 5-23, a 6.4% weight loss up to 100°C and endotherms at 86.0°C, 127.2°C and 133.1°C (peak) were observed. As shown in FIG. 5-24, no signal attributable to MeOH or MeTHF was observed in the 1H NMR spectrum of the solution. Based on the results, type G was likely a hydrate.

5.10. Type H.

Type H sample (3-10-A1) was prepared by adding antisolvent using acetone/H ₂ O. The XRPD result shown in FIG. 5-25, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-26, a 3.6% weight loss up to 130°C and an endotherm at 107.6°C (peak) were observed. As shown in FIG. 5-27, the presence of acetone was detected in the 1H NMR spectrum. Based on the results, type H was likely an acetone solvate.

5.11. Type I

Type I sample (3-10-A3) was prepared by antisolvent addition using DMSO/H2O. The XRPD result shown in FIG. 5-28, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-29, a 7.3% weight loss up to 150°C and an endotherm at 128.9°C (peak) were observed. As shown in FIG. 5-30, the presence of DMSO was detected in the 1H NMR spectrum. Based on the results, type I was likely a DMSO solvate.

5.12. Type J.

Sample type J (3-08-A2) was obtained by slow evaporation in THF. The XRPD result shown in FIG. 5-31, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-32, a 7.3% weight loss up to 125°C and an endotherm at 115.7°C (peak) were observed. As shown in FIG. 5-33, the presence of THF was detected in the ¹ H NMR spectrum. Based on the results, type J was likely a THF solvate.

5.13. Type K.

Sample type K (3-08-A14) was obtained by slow evaporation in acetone. The XRPD result shown in FIG. 5-34, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-35, a 5.7% weight loss up to 150°C and endotherms at 96.7°C, 119.8°C and 147.4°C (peak) and an ectotherm at 157.4°C were observed. As shown in FIG. 5-36, the presence of acetone was detected in the 1H NMR spectrum. Based on the results, Type K was likely an acetone solvate.

5.14. Type L

Type L sample (3-11-A4) was prepared by liquid-vapor diffusion in 2-MeTHF/nheptane. The XRPD result shown in FIG. 5-37 indicated a crystalline form with a preferred orientation. As shown by the TGA and DSC data in FIG. 5-38, a 7.9% weight loss up to 130°C and an endotherm at 126.2°C (peak) were observed. As shown in FIG. 5-39, the presence of acetone was detected in the 1H NMR spectrum, while no signal attributable to n-heptane was observed. Based on the results, type L was likely a solvate of 2-MeTHF.

5.15. Type M

Type M sample (3-11-A2) was prepared by liquid-vapor diffusion in EtOAc/IPA. The XRPD result shown in FIG. 5-40, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-41, a 3.7% weight loss up to 150°C and an endotherm at 122.6°C (peak) were observed. As shown in FIG. 5-42, the presence of IPA was detected in the 1H NMR spectrum, while no signal attributable to EtOAc was observed. Based on the results, Type M was likely an IPA solvate.

5.16. Type N.

Sample type N (3-08-A8) was obtained by slow evaporation in EtOH. The XRPD result shown in FIG. 5-43, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-44, a 4.1% weight loss up to 150°C and endotherms at 85.4°C, 126.5°C and 150.9°C (peak) were observed. As shown in FIG. 5-45, the presence of EtOH was detected in the 1H NMR spectrum. Based on the results, type M was likely a solvate with EtOH.

5.17. Type O

Type O sample (3-08-A11) was obtained by slow evaporation in MIBK. The XRPD result shown in FIG. 5-46, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-47, a 2.0% weight loss up to 130°C and two endotherms were observed at 106.2°C and 151.2°C (peak). As shown in FIG. 5-48, the presence of MIBK was detected in the 1H NMR spectrum. Based on the results, type O was likely a solvate of MIBK.

5.18. Type P

Sample type P (3-07-A14) was obtained by slow evaporation in DMF. The XRPD result shown in FIG. 5-49, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-50, a weight loss of 8.3% up to 130°C and an endotherm at 89.9°C (peak) were observed. As shown in FIG. 5-51, the presence of DMF was detected in the 1H NMR spectrum. Based on the results, type P was likely a solvate of DMF.

5.19. Type Q.

Type Q sample (3-11-A1) was prepared by liquid-vapor diffusion in MIBK/nheptane. The XRPD result shown in FIG. 5-52, indicated the crystalline form. As shown by the TGA and DSC data in FIG. 5-53, a 6.0% weight loss up to 120°C and endotherms at 92.9°C, 148.9°C and 170.0°C (peak) were observed. As shown in FIG. 5-54, the presence of MIBK was detected in the 1H NMR spectrum, while no signal attributable to n-heptane was observed. Based on the results, type Q was likely a solvate of MIBK.

6. Crystal data and experimental part for composition 4a.

Experimental part. Single colorless plate-like crystals (composition 4a) were used as received. A suitable crystal (0.28 x 0.18 x 0.09) mm ³ was selected and mounted on a nylon loop with paratonic oil on a Bruker APEX-II CCD diffractometer. The crystal was kept at T=173(2) K during data collection. Using Olex2 (Dolomanov et al., 2009), the structure was determined using the XT structure determination program (Sheldrick, 2015) using an internal phasing-based determination method. The model was refined using version XL (Sheldrick, 2008) using least squares minimization.

Data regarding the crystal. C ₆₅ H ₇₂ Cl ₂ F ₂ N ₈ O ₁₅ , M _r =1314.20, triclinic, P1 (No. 1), a=11.5683(10) A, b=11.6705(10) A, c= 13.9593(12) A, α=68.1780(10)°, β=69.4150(10)°, γ=87.7760(10)°, V=1628.7(2) A ³ , T =173(2) K, Z=1, Z'=1, μ(ΜοΚ _α )=0.178, 26758 reflections were measured, 11949 unique (R; _nt =0.0528), which were used for all calculations. The final wR ₂ was 0.2465 (all data) and R1 was 0.0835 (I > 2(1)).

- 63 045678

Table 6-2

Fractional atomic coordinates (x 10 ⁴ ) and equivalent isotropic displacement parameters (A ² x 10 ³ ) for composition 4A. U _eq is defined as 1/3 of the trace of the orthogonalized Ui,

Atom X U Z v _eq O1S 5607(5) 3451(5) 6493(5) 44.9(14) O2C 6723(6) 4580(6) 6921(5) 49.9(15) OZS 3704(5) 3216(6) 5826(5) 45.6(14) O4S 2800(6) 3841(6) 4560(6) 55.9(16) O5S 3369(5) 5630(6) 6567(5) 50.2(15) O6S 3636(5) 3990(6) 7917(5) 50.8(15) O7S 5703(6) 2984(7) 4257(5) 55.4(16) O8S 6595(5) 4867(6) 3832(5) 49.1(15) S1S 5032(7) 4527(8) 6031(7) 41(2) С2С 4615(7) 4267(8) 5203(7) 39.1(19) SZS 3928(8) 4667(8) 6968(7) 42(2) С4С 5705(8) 3954(9) 4361(7) 42(2) S5S 6393(8) 3601(8) 6961(7) 42(2) С6С 6820(9) 2403(8) 7493(8) 49(2) С7С 7721(15) 2378(12) 7941(13) 98(3) С8С 8179(15) 1311(13) 8410(13) 94(3) S9S 7697(17) 223(14) 8497(14) 107(5) С10С 6708(16) 185(12) 8178(13) 98(3) THX 6265(15) 1303(12) 7665(13) 94(3)

- 64 045678

CI2C 2953(8) 3028(9) 5341(7) 44(2) C13C 2359(9) 1762(9) 5860(8) 54(2) CMC 1223(9) 1501(10) 5822(8) 56(2) C15C 671(11) 300(12) 6312(10) 75(3) C16C 1261(14) -658(13) 6805(13) 98(5) C17C 2446(14) -407(13) 6720(13) 97(4) C18C 2954(13) 800(12) 6325(12) 87(4) CUB 6935(2) 1601(2) 11106(2) 66.0(7) FIB 4643(5) 1859(6) 10647(5) 66.7(16) O1B 8509(6) 7968(6) 7303(6) 58.0(17) O2B 4575(6) 6378(7) 8234(6) 66.1(19) NIB 7896(6) 6005(6) 8607(6) 40.4(16) N2B 7345(6) 3931(7) 9798(6) 42.2(17) N3B 6567(7) 7139(7) 7745(6) 49.2(18) N4B 11242(6) 5811(7) 8040(6) 47.4(18) C1B 7710(8) 7096(9) 7832(8) 48(2) C2B 7030(8) 4974(8) 9152(6) 40(2) C3B 6537(8) 2952(9) 10275(7) 47(2) C4B 5411(8) 2909(8) 10151(7) 47(2) C5B 5070(8) 3977(10) 9543(7) 49(2) C6B 5884(8) 5054(9) 9031(7) 44(2) C7B 5584(8) 6210(9) 8328(8) 50(2) C8B 9103(7) 5996(7) 8717(7) 41(2) C9B 9205(8) 6331(8) 9550(7) 43(2) C10B 10370(9) 6393(9) 9588(7) 49(2) CUB 11350(8) 6131(8) 8840(7) 44(2) C12B 10113(7) 5734(7) 7981(7) 38.9(19) C13B 8078(9) 6634(9) 10360(8) 55(2) C14B 10040(9) 5292(9) 7127(8) 54(2) C15B 10546(10) 4041(10) 7234(10) 65(3) C16B 10688(11) 6301(12) 5939(9) 73(3) CHA 11316(3) -588(3) 4003(3) 82.7(9) F1A 13799(6) -251(6) 4033(5) 77.9(18)

- 65 045678

O1A 12158(6) 6025(6) 1115(6) 57.3(17) O2A 15430(6) 4363(7) 1981(6) 63.4(18) NIA 11949(6) 3944(6) 2039(6) 38.9(16) N2A 11735(7) 1804(7) 3004(6) 48.6(19) N3A 13776(6) 5170(7) 1580(6) 45.5(18) N4A 8608(6) 4153(7) 2689(6) 46.4(18) CIA 12607(8) 5110(9) 1551(7) 47(2) C2A 12449(7) 2876(8) 2555(7) 40(2) C3A 12216(10) 797(10) 3464(8) 56(2) C4A 13389(10) 813(9) 3546(8) 56(3) C5A 14083(9) 1886(10) 3114(8) 54(2) C6A 13638(8) 2995(8) 2564(7) 44(2) C7A 14365(8) 4199(9) 2043(8) 48(2) C8A 10709(7) 3834(8) 2039(6) 37.4(19) C9A 10540(8) 3371(9) 1305(8) 47(2) C10A 9326(8) 3313(8) 1314(7) 47(2) C11A 8421(8) 3689(8) 2008(7) 45(2) C12A 9733(7) 4227(7) 2738(7) 39.4(19) C13A 11568(8) 2995(10) 522(8) 55(2) C14A 9890(8) 4703(9) 3549(7) 50(2) C15A 9619(11) 6056(11) 3255(11) 71(3) C16A 9045(10) 3875(11) 4734(8) 64(3) 03 s 355(10) 1070(10) 9718(9) 116(3) C.H.S. -300(40) -220(40) 11630(40) 142(14) CUT -1050(40) -700(50) 11410(40) 190(20) C12S 160(30) -310(30) 10410(20) 180(9) C13S 1360(40) -510(50) 10600(40) 151(11) C13T 1220(40) -820(40) 9990(40) 151(11) CMS 2240(20) 280(20) 9240(20) 155(8) C15S 1510(20) 1150(20) 8834(18) 139(7) 02S 6461(8) -721(8) 5995(7) 83(2) C6S 5780(20) -1920(20) 5275(19) 145(7) C7S 6000(20) -720(20) 5170(20) 155(8) C8S 7200(30) -180(30) 4150(30) 241(15) C9S 7990(20) 670(20) 4390(20) 153(7) CIOS 7490(30) 340(30) 5490(30) 202(11) 01S 4966(9) 7468(9) 1097(8) 99(3) CIS 5930(20) 8160(20) 2110(20) 176(9) C2S 5018(15) 7306(16) 2147(14) 105(5) C3S 3770(20) 7280(20) 2920(20) 146(7) C4S 3200(30) 8270(30) 2170(30) 207(12) C5S 4180(20) 8430(20) 990(20) 162(8)

- 66 045678

Table 6-3

Anisotropic displacement parameters (x 10 ⁴ ) for composition 4A

The exponential exponent of the anisotropic displacement coefficient has the following form:

-2l ² [th ² a* ² x Tsts+ ... +2hka* x b* χ υ _ί2 ]

Atom Up U22 and ₃₃ and ₂₃ Ui ₃ U12 O1S 37(3) 57(4) 52(3) -27(3) -25(3) 9(3) O2C 47(4) 54(4) 52(4) -21(3) -21(3) 4(3) OZS 33(3) 63(4) 39(3) -21(3) -10(3) -3(3) O4S 52(4) 68(4) 53(4) -20(4) -26(3) 1(3) O5S 34(3) 56(4) 53(4) -19(3) -11(3) 9(3) O6S 44(3) 71(4) 35(4) -21(3) -11(3) 3(3) O7S 46(4) 71(5) 56(4) -34(4) -15(3) 3(3) O8S 33(3) 56(4) 55(4) -22(3) -11(3) 7(3) S1S 31(4) 48(5) 47(5) -20(4) -14(4) 1(4) С2С 35(5) 42(5) 41(5) -16(4) -14(4) 2(4) SZS 37(5) 52(5) 39(5) -21(4) -14(4) 2(4) С4С 36(5) 51(5) 45(5) -17(4) -22(4) 6(4) S5S 40(5) 48(5) 35(4) -16(4) -11(4) 5(4) С6С 58(6) 45(5) 51(5) -22(4) -26(5) Yu(4) С7С 141(9) 68(6) 117(8) -37(6) -85(8) 23(6) С8С 124(9) 77(6) 120(8) -41(6) -86(7) 41(6) S9S 155(14) 87(10) 133(13) -53(9) -107(12) 68(10) С10С 141(9) 68(6) 117(8) -37(6) -85(8) 23(6) THX 124(9) 77(6) 120(8) -41(6) -86(7) 41(6) С12С 35(5) 65(6) 38(5) -26(5) -15(4) 3(4)

-67045678

C13C 50(6) 59(6) 56(6) -23(5) -23(5) 6(5) CMC 47(6) 71(7) 58(6) -31(5) -22(5) 1(5) C15C 54(6) 101(10) 73(8) -34(7) -22(6) -21(6) C16C 96(10) 81(9) 114(11) -3(8) -67(9) -27(8) C17C 98(10) 73(8) 110(11) -10(8) -51(9) -2(7) C18C 88(9) 77(8) 102(10) -20(7) -56(8) -9(7) CUB 67.9(16) 57.4(14) 64.2(16) -10.1(12) -27.3(13) -6.4(12) FIB 67(4) 76(4) 55(3) -19(3) -22(3) -23(3) O1B 41(4) 55(4) 71(4) -12(3) -23(3) -6(3) O2B 36(4) 89(5) 78(5) -31(4) -28(3) 11(3) NIB 23(3) 42(4) 51(4) -12(3) -13(3) -6(3) N2B 33(4) 54(5) 35(4) -14(3) -10(3) -5(3) N3B 40(4) 58(5) 48(4) -12(4) -23(4) 2(4) N4B 35(4) 47(4) 59(5) -17(4) -18(4) 7(3) C1B 38(5) 55(6) 58(6) -27(5) -22(4) 9(5) C2B 37(5) 53(5) 27(4) -15(4) -9(4) 1(4) C3B 46(5) 55(6) 40(5) -19(4) -14(4) 6(4) C4B 49(5) 54(6) 36(5) -16(4) -14(4) -12(4) C5B 29(5) 83(7) 35(5) -25(5) -9(4) -6(5) C6B 32(5) 64(6) 37(5) -21(4) -10(4) -8(4) C7B 38(5) 67(6) 50(5) -23(5) -19(4) 3(4) C8B 34(5) 43(5) 48(5) -15(4) -19(4) 2(4) C9B 39(5) 51(5) 42(5) -22(4) -12(4) 1(4) C10B 49(6) 59(6) 42(5) -20(4) -20(4) 10(4) CUB 42(5) 50(5) 46(5) -17(4) -23(4) 4(4) C12B 28(4) 42(5) 47(5) -17(4) -14(4) 2(3) C13B 44(5) 62(6) 52(6) -25(5) -8(4) 7(5) C14B 40(5) 62(6) 61(6) -26(5) -18(5) 0(4) C15B 52(6) 72(7) 89(8) -52(6) -22(6) 7(5) C16B 66(7) 99(9) 50(6) -23(6) -21(5) -10(6) CHA 84(2) 60.8(16) 85(2) -4.6(15) -34.1(16) -3.2(14) F1A 89(5) 73(4) 76(4) -21(3) -45(4) 31(3) O1A 45(4) 60(4) 64(4) -17(4) -23(3) 1(3)

- 68 045678

O2A 42(4) 87(5) 76(5) -38(4) -33(3) 8(3) N1A 26(3) 53(4) 37(4) -16(3) -12(3) 1(3) N2A 41(4) 59(5) 37(4) -13(4) -9(3) 0(4) N3A 31(4) 58(4) 51(4) -27(4) -12(3) -4(3) N4A 36(4) 59(5) 40(4) -16(4) -12(3) 1(3) C1A 44(5) 57(6) 41(5) -21(4) -16(4) 2(5) C2A 30(4) 65(6) 33(4) -26(4) -13(4) 12(4) SZA 61(6) 65(6) 48(6) -20(5) -28(5) 6(5) C4A 57(6) 55(6) 60(6) -29(5) -19(5) 18(5) C5A 54(6) 79(7) 48(5) -36(5) -27(5) 18(5) C6A 34(5) 59(6) 45(5) -26(4) -15(4) 13(4) C7A 35(5) 72(6) 47(5) -34(5) -13(4) 5(5) C8A 32(4) 46(5) 30(4) -10(4) -12(4) 6(4) C9A 33(5) 61(6) 48(5) -18(4) -16(4) 5(4) S10A 38(5) 59(6) 46(5) -17(4) -18(4) -5(4) SNA 30(5) 59(6) 38(5) -14(4) -8(4) -1(4) S12A 31(4) 47(5) 34(4) -9(4) -Yu(4) 3(4) C13A 39(5) 91(7) 41(5) -34(5) -Yu(4) 12(5) S14A 36(5) 73(6) 46(5) -31(5) -12(4) 5(4) S15A 64(7) 81(8) 78(8) -44(6) -22(6) 6(6) S16A 58(6) 96(8) 39(5) -21(5) -25(5) 16(6) O3S 113(8) 120(8) 98(7) -25(6) -37(6) 19(7)

Atom Atom Length/A O1S S1S 1.439(10) O1S S5S 1,341(10) O2C S5S 1,194(10) OZS С2С 1.427(10) ozs С12С 1,341(10) O4S С12С 1.215(11) O5S sss 1.313(10) O6S sss 1.193(10) O7S С4С 1,194(10)

Table 6-3

Bond lengths, expressed in A, for composition 4A

Atom Atom Length/A O8S С4С 1.306(10) S1S С2С 1.525(11) S1S SZS 1.528(12) С2С С4С 1.532(12) S5S С6С 1,478(13) С6С С7С 1,386(16) С6С THX 1,358(16) С7С С8С 1,358(17) С8С S9S 1.35(2)

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C9C C10C 1,373(19) sus cue 1,411(17) C12C C13C 1.454(14) C13C C14C 1,384(13) C13C C18C 1.366(16) C14C C15C 1.375(15) C15C C16C 1.375(18) C16C C17C 1,368(19) C17C C18C 1.375(18) CUB C3B 1.732(10) FIB C4B 1.339(10) 01B C1B 1.212(11) O2B C7B 1.221(11) NIB C1B 1,398(12) NIB C2B 1,381(10) NIB C8B 1.458(10) N2B C2B 1,344(11) N2B C3B 1.305(11) N3B C1B 1.367(11) N3B C7B 1.385(12) N4B CUB 1.346(11) N4B C12B 1.346(10) C2B C6B 1.391(12) C3B C4B 1.378(13) C4B C5B 1.357(13) C5B C6B 1,389(12) C6B C7B 1.456(14) C8B C9B 1,400(12) C8B C12B 1.369(12) C9B C10B 1.372(12) C9B C13B 1.526(13) C10B CUB 1.359(13) C12B C14B 1,492(13)

C14B C15B 1.526(14) C14B C16B 1,562(15) CHA C3A 1.714(11) F1A C4A 1.336(11) O1A CIA 1.216(11) O2A C7A 1.223(11) NIA CIA 1.386(11) NIA C2A 1.405(11) NIA C8A 1.446(10) N2A C2A 1.331(11) N2A C3A 1.321(12) N3A CIA 1.373(12) N3A C7A 1.368(12) N4A CHA 1.335(11) N4A C12A 1.335(11) C2A C6A 1,394(12) C3A C4A 1,402(14) C4A C5A 1.324(14) C5A C6A 1.419(13) C6A C7A 1.453(13) C8A C9A 1.393(12) C8A C12A 1,398(12) C9A C10A 1.405(12) C9A C13A 1.485(13) C10A C11A 1.339(13) C12A C14A 1.497(12) C14A C15A 1.527(16) C14A C16A 1,541(14) 03 s C12S 1.51(3) 03 s C15S 1.44(2) C.H.S. C12S 1.64(5) CUT C12S 1.51(3) C12S CBS 1.49(5)

-70045678

- 71 045678

Table 6-4

Bond angle values for composition 4A

Atom Atom Atom Angle/ ⁰ S5S O1S S1S 116.5(6) С12С ozs С2С I7.1(7) O1S S1S С2С 105.3(6) O1S S1S SZS 108.6(7) sss S1S С2С 111.5(7) ozs С2С S1S 106.6(6) ozs С2С С4С 108.9(6) S1S С2С С4С 110.9(7) O5S SZS S1S 109.9(7) O6S SZS O5S 126.1(8) O6S sss S1S 124.0(8) O7S С4С O8S 127.3(8) O7S С4С С2С 122.3(8) O8S С4С С2С 110.4(7) O1S S5S С6С 110.8(8) O2C S5S O1S 124.1(8) O2C S5S С6С 125.1(8) С7С С6С S5S 119.6(9) С11С С6С S5S 121.9(9) С11С С6С С7С 118.1(10) С8С С7С С6С 122.4(12) S9S С8С С7С 119.1(13) С10С S9S С8С 120.6(12) S9S С10С THX 119.5(13) С6С THX С10С 119.7(12) OZS С12С С13С 112.4(8) O4S С12С OZS 122.7(8) O4S С12С С13С 124.9(8) С14С С13С С12С 120.6(9) С18С С13С С12С 120.4(9) С18С С13С С14С 118.8(10) С15С С14С С13С 120.1(10)

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C16C С15С С14С 120.5(10) C15C С16С С17С 118.7(12) C16C С17С С18С 120.4(14) C13C С18С С17С 120.5(12) C1B N1B С8В 115.5(7) C2B N1B S1B 121.7(7) C2B N1B С8В 122.6(7) SZV N2B C2B 116.6(7) C1B N3B S7V 126.9(8) С12В N4B SIV 118.8(8) 01B S1B N1B 121.2(8) 01B S1B N3B 122.4(9) N3B S1B N1B 116.3(8) N1B C2B С6В 119.7(8) N2B C2B N1B 117.3(7) N2B C2B С6В 123.1(8) N2B SZV START 116.9(7) N2B SZV С4В 125.0(8) С4В SZV S11V 118.0(7) F1B С4В SZV 121.5(8) F1B С4В S5V 120.2(8) S5V С4В SZV 118.3(8) С4В S5V S6V 119.2(8) C2B С6В S7V 120.9(8) S5V С6В C2B 117.7(8) S5V S6V S7V 121.2(8) 02B S7V N3B 121.3(9) O2B S7V С6В 124.9(9) N3B S7V С6В 113.8(7) S9V С8В N1B 118.0(7) С12В С8В N1B 120.2(7) С12В С8В S9V 121.7(7) С8В S9V С13В 121.4(8)

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suv C9B C8B 116.6(8) suv C9B C13B 122.0(8) CUB C10B C9B 119.9(8) N4B CUB C10B 123.0(8) N4B C12B C8B 119.9(8) N4B C12B C14B 116.2(7) C8B C12B C14B 123.8(7) C12B C14B C15B 113.0(8) C12B C14B C16B 110.5(8) C15B C14B C16B 111.7(9) CIA NIA C2A 121.5(7) CIA NIA C8A 118.9(7) C2A NIA C8A 119.5(7) C3A N2A C2A 116.6(8) C7A N3A CIA 126.9(8) C12A N4A C11A 120.6(7) 01A CIA NIA 120.7(8) O1A CIA N3A 122.4(8) N3A CIA NIA 116.9(8) N2A C2A NIA 116.6(7) N2A C2A C6A 124.3(8) C6A C2A NIA 119.1(8) N2A C3A CHA 116.6(7) N2A C3A C4A 123.7(9) C4A C3A CHA 119.7(8) F1A C4A C3A 119.7(9) C5A C4A F1A 121.1(9) C5A C4A C3A 119.2(9) C4A C5A C6A 119.7(9) C2A C6A C5A 116.4(9) C2A C6A C7A 121.0(8) C5A C6A C7A 122.6(8) O2A C7A N3A 120.9(9)

- 74 045678

O2A C7A C6A 124.4(9) N3A C7A C6A 114.7(8) C9A C8A NIA 118.0(7) C9A C8A C12A 122.2(7) CI2A C8A NIA 119.7(7) C8A C9A C10A 115.9(8) C8A C9A C.B.A. 123.3(8) C10A C9A C.B.A. 120.7(8) SNA C10A C9A 119.5(8) C10A CHA N4A 123.5(8) N4A C12A C8A 118.2(8) N4A C12A CMA 118.8(7) C8A C12A CMA 123.0(7) C12A C14A C15A 110.2(8) C12A C14A C16A 109.8(8) C15A C14A C16A 111.6(9) C15S 03 s C12S 101.0(16) 03 s C12S C.H.S. 98(2) sit C12S 03 s 114(3) CBS C12S 03 s 101(3) CBS C12S CUT 119(4) CBT C12S 03 s 108(3) CBT C12S C.H.S. 124(4) C12S CBS CMS 94(3) C12S CBT CMS 106(3) C15S CMS CBS 105(2) C15S CMS CBT 98(2) CMS C15S 03 s 109.6(18) C7S 02S CIOS 109.8(17) 02S C7S C8S 98(2) C6S C7S 02S 110(2) C6S C7S C8S 105(2) C7S C8S C9S 108(3) CIOS C9S C8S 104(3) C9S CIOS 02S 110(2) C5S 01S C2S 99.3(14) 01S C2S CIS 115.3(16) 01S C2S C3S 109.3(14) CIS C2S C3S 113.3(18) C2S C3S C4S 102.4(19) C3S C4S C5S 100(2) 01S C5S C4S 110(2)

- 75 045678

Table 6-5

Fractional coordinates of hydrogen atoms (x 10 ⁴ ) and parameters of the equivalent isotropic displacement (A ² x 10 ³ ) for composition 4A. Ue, defined as 1/3 of the trace orthogonalized, Uij

Atom X U Z u _eq H5S 2738 5676 7079 75 H8S 7257 4612 3523 74 H1S 5642 5286 5648 49 H2S 4246 5000 4813 47 H7S 8030 3133 7918 117 N8SA 8829 1329 8674 FROM H9S 8046 -520 8781 129 H10S 6325 -589 8302 117 NIS 5582 1286 7440 FROM H14S 824 2153 5456 67 H15S -125 131 6310 90 H16S 854 -1479 7197 118 H17S 2922 -1070 6936 117 Н18С 3724 966 6375 104 NZV 6442 7830 7268 59 Н5В 4284 3988 9467 59 Н10В 10491 6619 10136 58 NIV 12147 6175 8883 53 H13A 7637 7233 9947 82 Н13В 8351 6990 10789 82

- 76 045678

H13C 7521 5873 10864 82 H14B 9141 5177 7246 64 H15A 11415 4105 7168 98 H15B 10492 3806 6645 98 H15D 10055 3410 7957 98 H16A 10303 7074 5885 109 H16B 10596 6013 5392 109 H16D 11573 6448 5797 109 NZA 14191 5912 1268 55 H5A 14875 1914 3171 65 H10A 9147 3010 831 57 NIA 7601 3622 2016 54 H13D 12178 3715 8 83 H13E 11236 2674 104 83 H13F 11970 2347 938 83 H14A 10773 4660 3501 60 H15E 9817 6386 3737 106 H15F 8738 6107 3360 106 H15G 10127 6543 2480 106 H16E 8174 3966 4816 96 H16F 9227 4126 5267 96 H16G 9198 3006 4874 96 Hl ID 403 121 11721 212 HUE -600 -1054 12212 212 Hl IF -968 318 11703 212 HUG -1060 -259 11882 290 HUH -1114 -1599 11825 290 Hill -1750 -509 11154 290 H12S 128 -753 9931 215 H12A -557 -686 10345 215 H13G 1497 -1393 10872 182 H13H 1451 -115 11089 182 H13I 1087 -1267 9548 182

-77045678

H13J 1458 -1412 10592 182 H14D 3019 696 9168 186 H14E 2460 -284 8839 186 H14F 2628 537 9673 186 H14G 2896 104 8645 186 N15N 1338 997 8243 167 H15I 1947 1996 8512 167 H6SA 6575 -2283 5108 218 H6SB 5393 -1922 4756 218 H6SC 5233 -2417 6034 218 H7S 5282 -222 5136 186 H8SA 7003 307 3490 290 H8SB 7680 -861 4007 290 H9SA 8880 527 4152 184 H9SB 7923 1561 4003 184 H10D 7155 1055 5667 243 H10E 8144 66 5812 243 Hl SA 6626 7716 2249 264 Hl SB 5541 8496 2685 264 H1SC 6226 8839 1383 264 H2S 5277 6457 2441 126 H3SA 3789 7506 3528 175 H3SB 3293 6455 3231 175 H4SA 3168 9048 2302 249 H4SB 2364 7965 2275 249 H5SA 3745 8410 500 195 H5SB 4689 9246 647 195

Table 6-4

Hydrogen Bonding Information for Composition 4A

D H A d(D-H)/A d(H-A)/A d(D-A)/A D-H-A/deg. O5C H5C N4B ¹ 0.84 1.82 2.656(9) 170.9 O8C H8C N4A 0.84 1.79 2.624(9) 171.6 N3B H3B O2S ² 0.88 1.94 2,805(12) 168.1 C14B H14B NIB 1.00 2.44 2.937(12) 110.1 N3A H3A O1S ³ 0.88 1.95 2,798(12) 160.7

¹ - 1+x,+y,+z; ² +x,1+y,+z; ³ 1+x,+y,+z

- 78 045678

The foregoing is only illustrative of the present invention and is not intended to limit the present invention to the disclosed uses. It is intended that variations and modifications that are standard to those skilled in the art are within the scope and spirit of the present invention, as defined in the appended claims. All references, patents, applications and publications mentioned herein are hereby incorporated by reference in their entirety to the extent described herein.

Claims (54)

1. Composition for preparing a compound of formula 5M having the structure wherein the composition contains a compound of formula 4 and a compound of formula B 2. The composition of claim 1, wherein the compound of formula 4 is a compound of formula 5M - 79 045678 3. The composition according to claim 1, wherein the compound of formula 4 is a compound of formula 5P o 4. Composition according to any one of claims 1 to 3, wherein the compound of formula B is a compound of formula B1 Ph Vo co ₂ no U-( o NO ₂ C 0^ Ph Bl. 5. Composition according to any one of claims 1 to 3, wherein the compound of formula B is a compound of formula B2 Ph Vo co ₂ no )—{ o no _g s Ph B2. 6. The composition according to any one of claims 1 to 5, wherein the composition comprises a 2 to 1 ratio of the compound of formula 4 to the compound of formula B. 7. The composition according to any one of claims 1 to 5, where the composition additionally contains 2methyltetrahydrofuran. 8. The composition according to claim 7, wherein the ratio of 2-methyltetrahydrofuran and the compound of formula B is 2 to 1. 9. The composition according to claim 1, where the composition is 10. The composition according to claim 9, where the composition is 11. The composition according to claim 9, where the composition is 12. The composition according to any one of claims 1-11, where the composition is in a crystalline state. 13. The composition according to any one of claims 1 to 11, where the composition is a co-crystal. 14. Method for preparing the composition of formula 4a comprising mixing a compound of formula 4 - 80 045678 with compound of formula B1 in the presence of 2-methyltetrahydrofuran to form a composition of formula 4a. 15. The method of claim 14, wherein 1.0 molar equivalents of the compound of formula 4 are mixed with 2.0 molar equivalents of the compound of formula B1. 16. The method according to claim 14 or 15, wherein the compound of formula 4 and the compound of formula B1 are mixed in a solvent, wherein the solvent contains 2-methyltetrahydrofuran and heptane. 17. Method for preparing the compound of formula 5M comprising mixing a composition of formula 4a with a base to obtain the compound of formula 5M. 18. The method according to claim 17, where the base is Na ₂ HPO ₄ . 19. The method according to claim 17, where the base is NaHCO ₃ . 20. The method according to any one of claims 17 to 19, wherein 1.0 molar equivalents of the composition of formula 4a are mixed with 2.0 molar equivalents of base. 21. The method according to any one of claims 17 to 20, wherein the composition of formula 4a is mixed with a base in a solvent, wherein the solvent contains methyl tert-butyl ether and water. 22. Method for preparing the compound of formula 9 or a pharmaceutically acceptable salt thereof, comprising mixing 6-fluoro-7-(2-fluoro-6hydroxyphenyl)-4-[(2S)-2-methylpiperazin-1-yl]-1-[4-methyl-2-(propan- 2-yl)pyridin-3-yl]pyrido[2,3d]pyrimidin-2(1H)-one with acryloyl chloride to form 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4methyl-2 -(propan-2-yl)pyridin-3-yl]-4-[(2S)-2-methyl-4-(prop-2-enoyl)piperazin-1-yl]pyrido[2,3d]pyrimidin-2 (1H)-one, where 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperazin-1-yl]-1-[4methyl-2-(propan-2 -yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one is stereoisomerically enriched in 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S) -2-methylpiperazin-1-yl]-(1M)-1-[4-methyl-2-(propan2 -yl)pyridin-3 -yl]pyrido[2,3-d]pyrimidin-2 (III)-one . 23. The method according to claim 22, where 1.0 molar equivalent of 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2methylpiperazin-1-yl]-1-[4-methyl -2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one is mixed with 1.3 molar equivalents of acryloyl chloride. 24. The method according to claim 22 or 23, where 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2-methylpiperazin-1-yl]1-[4-methyl-2 -(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one and acryloyl chloride are mixed in a solvent, where the solvent contains N-methylpyrrolidone. - 81 045678 25. The method according to any one of claims 22-24, where 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2methylpiperazin-1-yl]-1-[4-methyl- 2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one is essentially stereoisomerically pure 6-fluoro-7-(2-fluoro-6- Hydroxyphenyl)-4-[(2S)-2methylpiperazin-1 -yl]-(1M)-1 -[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d ]pyrimidin-2(1H)-one. 26. The method according to any one of claims 22-25, where the production of 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2methylpiperazin-1-yl]-1-[4-methyl -2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one involves mixing tert-butyl (3S)-4-{6-fluoro-7- (2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2-(propan-2yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4 -yl}-3-methylpiperazin-1-carboxylate and trifluoroacetic acid to form 6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-[(2S)-2methylpiperazin-1-yl]-1-[4 -methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2(1H)-one, where tert-butyl (3S)-4-{6-fluoro- 7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo1,2-dihydropyrido[2,3-d]pyrimidine -4-yl}-3-methylpiperazine-1-carboxylate is stereoisomerically enriched in tert-butyl(3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-(1M)-1-[ 4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazin-1 carboxylate. 27. The method according to claim 26, where tert-butyl (3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2(propan-2-yl) pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1carboxylate and trifluoroacetic acid are mixed in a solvent, where the solvent contains dichloromethane. 28. The method according to claim 26 or 27, where tert-butyl (3S)-4-{6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2(propane-2- yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1carboxylate is essentially stereoisomerically pure tert-butyl (3S)- 4-{6-fluoro-7-(2-fluoro-6hydroxyphenyl)-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1, 2-dihydropyrido[2,3d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate. 29. The method according to any one of claims 26-28, where the production of tert-butyl (3S)-4-{6-fluoro-7-(2-fluoro-6hydroxyphenyl)-1-[4-methyl-2-(propane- 2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin4-yl}-3-methylpiperazine-1-carboxylate involves mixing tert-butyl (3S)-4- {7-chloro-6-fluoro-1-[4methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4- yl}-3methylpiperazine-1-carboxylate, potassium trifluoro(2-fluoro-6-hydroxyphenyl)borate, [1,1'bis(diphenylphosphino)ferrocene]dichloropalladium(P) and potassium acetate to form tert-butyl (3S)-4 {6-fluoro-7-(2-fluoro-6-hydroxyphenyl)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]-2-oxo-1,2dihydropyrido[2, 3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate, where tert-butyl (3S)-4-{7chloro-6-fluoro-1-[4-methyl-2-(propane-2- yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4yl}-3-methylpiperazine-1-carboxylate is a stereoisomerically enriched tert-butyl (3S)-4-{ 7-chloro6-fluoro-( 1M)-1 -[4-methyl-2-(propan-2-yl)pyridin-3 -yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidine -4yl}-3-methylpiperazine-1-carboxylate. 30. The method according to claim 29, where 1.0 molar equivalent of tert-butyl (3S)-4-{7-chloro-6-fluoro-1-[4-methyl-2(propan-2-yl)pyridin-3 -yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1carboxylate is mixed with 1.2 molar equivalents of trifluoro(2-fluoro-6-hydroxyphenyl) borate potassium 31. The method according to claim 29 or 30, where 1.0 molar equivalent of tert-butyl (3S)-4-{7-chloro-6-fluoro-1-[4methyl-2-(propan-2-yl)pyridin- 3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3methylpiperazine-1-carboxylate is mixed with 0.01 molar equivalent of [1,1'bis(diphenylphosphino)ferrocene ]dichloropalladium(P). 32. Method according to any one of claims 29-31, where 1.0 molar equivalent of tert-butyl (3S)-4-{7-chloro-6-fluoro1-[4-methyl-2-(propan-2-yl) Pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3methylpiperazine-1-carboxylate is mixed with 5.0 molar equivalents of potassium acetate. 33. Method according to any one of claims 29-32, where tert-butyl (3S)-4-{7-chloro-6-fluoro-1-[4-methyl-2-(propan-2yl)pyridin-3-yl ]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate, potassium trifluoro(2-fluoro-6-hydroxyphenyl)borate, [1,1 '-bis(diphenylphosphino)ferrocene]dichloropalladium(II) and potassium acetate are mixed in a solvent, where the solvent contains dioxane. 34. Method according to any one of claims 29-33, where tert-butyl (3S)-4-{7-chloro-6-fluoro-1-[4-methyl-2-(propan-2yl)pyridin-3-yl ]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1-carboxylate is essentially stereoisomerically pure tert-butyl (3S)-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-1carboxylate. 35. The method according to any one of claims 29-34, where the production of tert-butyl (3S)-4-{7-chloro-6-fluoro-1-[4-methyl-2(propan-2-yl)pyridin-3 -yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl}-3-methylpiperazine-1carboxylate involves mixing 4,7-dichloro-6-fluoro-1-(2-isopropyl -4-methylpyridin-3yl)pyrido[2,3-d]pyrimidin-2(1H)-one with tert-butyl (3S)-3-methylpiperazine-1-carboxylate and N,Ndiisopropylethylamine to form tert-butyl (3S) -4-{7-chloro-6-fluoro-1-[4-methyl-2-(propan-2yl)pyridin-3-yl]-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidine -4-yl}-3-methylpiperazine-1-carboxylate, - 82 045678 where 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3^]pyrimidin-2(III)-one is stereoisomerically enriched 4,7 -dichloro-6-fluoro-(1M)-1 -(2-isopropyl-4-methylpyridin-3 yl)pyrido[2,3^]pyrimidine-2(III)-om. 36. The method according to claim 35, where one molar equivalent of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)- it is mixed with 1.1 molar equivalent of tert-butyl (3S)-3-methylpiperazine-1-carboxylate. 37. The method according to claim 35 or 36, where one molar equivalent of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H ) - it is mixed with 1.0 molar equivalent of N,Ndiisopropylethylamine. 38. Method according to any one of claims 35-37, where 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3yl)pyrido[2,3-d]pyrimidin-2(1H) -one, tert-butyl (3S)-3-methylpiperazine-1-carboxylate and N,Ndiisopropylethylamine are mixed in a solvent, where the solvent contains toluene. 39. Method according to any one of claims 35-38, where 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3yl)pyrido[2,3-d]pyrimidin-2(1H) -one is essentially stereoisomerically pure 4,7-dichloro-6fluoro-(1M)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)- He. 40. The method according to any one of claims 35-39, where the production of 4,7-dichloro-6-fluoro-1-(2-isopropyl-4methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H )-one involves mixing 7-chloro-6-fluoro-1-[4-methyl2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidine-2,4(1H,3H )-dione with POCl ₃ and N,N-diisopropylethylamine to give 4,7-dichloro-6-fluoro-1-(2-isopropyl-4-methylpyridin-3yl)pyrido[2,3-d]pyrimidine-2(1H )-one, where 7-chloro-6-fluoro-1-[4-methyl-2-(propan-2-yl)pyridin-3yl]pyrido[2,3-d]pyrimidine-2,4(1H,3H )-dione is stereoisomerically enriched with 7-chloro-6-fluoro(1M)-1 - [4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin-2 ,4(1 H,3N)-dione. 41. The method according to claim 40, where one molar equivalent of 7-chloro-6-fluoro-1-[4-methyl-2-(propan-2yl)pyridin-3-yl]pyrido[2,3-d]pyrimidin- 2,4(1H,3H)-diones are mixed with 1.2 molar equivalents of POCl3. 42. The method according to claim 40 or 41, where one molar equivalent of 7-chloro-6-fluoro-1-[4-methyl-2-(propan-2yl)pyridin-3-yl]pyrido[2,3-d] Pyrimidine-2,4(1H,3H)-dione is mixed with 2.0 molar equivalents of N,N-diisopropylethylamine. 43. Method according to any one of claims 40-42, where 7-chloro-6-fluoro-1-[4-methyl-2-(propan-2-yl)pyridin-3yl]pyrido[2,3-d]pyrimidine -2,4(1H,3H)-dione, _POC13 and N,N-diisopropylethylamine are mixed in a solvent, where the solvent contains toluene. 44. Method according to any one of claims 40-43, where 7-chloro-6-fluoro-1-[4-methyl-2-(propan-2-yl)pyridin-3yl]pyrido[2,3-d]pyrimidine -2,4(1H,3H)-dione is essentially stereoisomerically pure 7chloro-6-fluoro-(1M)-1-[4-methyl-2-(propan-2-yl)pyridin-3-yl]pyrido [2,3-d]pyrimidine-2,4(1H,3H)-dione. 45. Composition for preparing a compound of formula 5M having the structure wherein the composition contains a compound of formula 4 and a compound of formula 11 46. The composition of claim 45, wherein the compound of formula 4 is a compound of formula 5M 47. The composition according to claim 45, wherein the compound of formula 4 is a compound of formula 5P - 83 045678 48. The composition according to any one of claims 45-47, where the compound of formula 11 is a compound of formula 11b hoJLo _L s o 11b. 49. A composition according to any one of claims 45 to 47, wherein the compound of formula 11 is a compound of formula 11a 50. The composition according to claim 45, where the composition is 51. The composition according to claim 45, where the composition is 52. The composition according to claim 45, where the composition is 53. The composition according to claim 45, where the composition is 54. The composition according to any one of claims 45 to 53, wherein the composition comprises a 1 to 1 ratio of the compound of formula 4 to the compound of formula 11.