Pyridopyrimidine derivatives useful as KRAS G12C and KRAS G12D inhibitors in the treatment of cancer

Abstract

Provided herein are KRAS G12C and KRAS G12D inhibitors, compositions of the same, and methods of using the same, which are useful for treating a number of disorders, including pancreatic cancer, colorectal cancer, and lung cancer. TIFF2022552873000118.tif44170

Description

[Technical field]

[0001] The present invention relates to compounds that inhibit the KRAS G12C and KRAS G12D proteins; methods of using the compounds to treat diseases or conditions, such as cancer; and pharmaceutical compositions containing the compounds. [Background technology]

[0002] 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 about 13% of NSCLC patients, and some studies have shown that KRAS mutations are a negative prognostic factor for NSCLC patients. Recently, V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations have been found to confer resistance to epidermal growth factor receptor (EGFR)-targeted therapy in colorectal cancer; therefore, the mutation status of KRAS can provide important information before prescribing TKI therapy. Summary of the Invention [Problem to be solved by the invention]

[0003] Taken together, there is a need for new medical treatments for patients with pancreatic cancer, lung adenocarcinoma or colorectal cancer, particularly those diagnosed with such cancers characterized by KRAS mutations and those who have progressed after chemotherapy.

[0004] The compounds disclosed herein may be in the form of pharma- ceutically acceptable salts.The compounds provided herein may be formulated into pharmaceutical compositions comprising the compounds disclosed herein and pharma- ceutically acceptable excipients.

[0005] Also provided is a method of inhibiting KRAS G12C in a cell, comprising contacting the cell with a compound or composition disclosed herein. Also provided is a method of inhibiting KRAS G12D in a cell, comprising contacting the cell with a compound or composition disclosed herein. KRAS G12D mutation arises from a single nucleotide change (c.35G>A), resulting in an amino acid substitution of glycine (G) at position 12 with aspartic acid (D). KRAS G12D is present in 3.99% of AACR GENIE cases, including colorectal adenocarcinoma, exocrine pancreatic neoplasm, non-small cell lung cancer, uterine neoplasm, and ovarian neoplasm, which have the highest prevalence. Also provided is a method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a compound or composition disclosed herein. In some embodiments, the cancer is lung cancer, pancreatic cancer, or colorectal cancer. [Means for solving the problem]

[0006] One aspect of the present invention provides the various compounds, stereoisomers, atropisomers, pharma- ceutically acceptable salts, pharma- ceutically acceptable salts of stereoisomers, and pharma- ceutically acceptable salts of atropisomers, as described in the embodiments below.

[0007] Another aspect of the invention provides a pharmaceutical composition comprising a compound of any of the embodiments, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier or diluent.

[0008] Another aspect of the invention provides a method of treating cancer. Such methods include: administering to a patient in need thereof a therapeutically effective amount of a compound of any of the embodiments or a pharma- ceutically acceptable salt thereof. In some such methods, the cancer is a hematological malignancy. In some such methods, the cancer is selected from the group consisting of breast cancer, colorectal cancer, skin cancer, melanoma, ovarian cancer, renal cancer, lung cancer, non-small cell lung cancer, lymphoma, non-Hodgkin's lymphoma, myeloma, multiple myeloma, leukemia, and acute myeloid leukemia. In some other such methods, the cancer is multiple myeloma. In some other such methods, the cancer is acute myeloid leukemia. In some other such methods, the cancer is non-Hodgkin's lymphoma. In another aspect, the method further includes administering to a patient in need thereof a therapeutically effective amount of an additional pharma- ceutically active compound. For example, in some such methods, the additional pharma- ceutically active compound is pembrolizumab. In other cases, the additional pharma- ceutically active compound is nivolumab. In yet other such methods, the additional pharma- ceutically active compound is trametinib. In yet other such methods, the additional pharma- ceutically active compound is RMC-4630. In yet other such methods, the additional pharma- ceutically active compound is AMG 404.

[0009] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Methods and materials are described herein for use in this disclosure; other suitable methods and materials known in the art can also be used. Materials, methods and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0010] Other features and advantages of the present disclosure will be apparent from the following detailed description and claims. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] definition Abbreviation: The following abbreviations may be used herein:

[0012] [Table 1]

[0013] [Table 2]

[0014] In connection with the description of the present invention (particularly in connection with the claims), the use of the terms "a", "an", "the" and similar referents should be construed to encompass both the singular and the plural unless otherwise indicated. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated herein as if it were individually recited herein. The use of any examples or exemplary language provided herein (e.g., "etc.") is intended to better illustrate the invention and is not a limitation on the scope of the invention unless otherwise asserted. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0015] The term "alkyl" as used herein includes, but is not limited to, straight-chain and branched C alkyl groups, including methyl, ethyl, n-propyl, i-propyl, n-butyl, sec-butyl, t-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, and 2-ethylbutyl. 1 ~C 8 Refers to a hydrocarbon group.m~n The term "alkylene" refers to an alkyl group having a substituent. The term "alkylene" refers to an alkyl group having an alkyl (e.g., methyl) group or an alkylene (e.g., -CH 2 -) groups include, for example, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, -NC, amino, -CO 2 H, -CO 2 C 1 ~C 6 Alkyl, -OCOC 1 ~C 6 Alkyl, C 3 ~C 10 Cycloalkyl, C 3 ~C 10 Heterocycloalkyl, C 5 ~C 10 Aryl and C 5 ~C 10 Heteroaryl may be substituted with one or more, typically 1 to 3, independently selected from the group consisting of aryl, heteroaryl, ...

[0016] The terms "alkenyl" and "alkynyl" refer to alkyl groups that further contain double or triple bonds, respectively.

[0017] The term "halo" as used herein refers to fluoro, chloro, bromo, and iodo. The term "alkoxy" is defined as -OR, where R is alkyl.

[0018] As used herein, the terms "amino" or "amine" refer interchangeably to -NR 2 refers to the group, where each R is, for example, H or a substituent group. In some embodiments, the amino group is further substituted to an ammonium ion, e.g., NR 3+The ammonium moiety is specifically included in the definition of "amino" or "amine". The substituents can be, for example, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, amido, or carboxylate. The R group can be further substituted with one or more, e.g., 1 to 4, groups selected from, for example, halo, cyano, alkenyl, alkynyl, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl, carboxylate, amine, and amide. An "amide" or "amido" group refers interchangeably to a group similar to an amine or amino group, but which does not include a C(O) group, e.g., -C(O)NR 2 Some possible amino or amido groups (some of which have an optional alkylene group, e.g., alkylene-amino or alkylene-amido) are CH 2 NH 2 , CH(CH 3 )NH 2 , CH(CH 3 ) 2 NH 2 , C.H. 2 CH 2 NH 2 , C.H. 2 CH 2 N(CH 3 ) 2 , C.H. 2 NHCH 3 , C(O)NHCH 3 , C(O)N(CH 3 ) 2 , C.H. 2 C(O)NHphenyl, CH 2 NHC(O)CH 3 , C.H. 2 NHCH 2 CH 2 OH, CH 2 NHCH 2 CO 2 H, C.H. 2 NH(CH 3 )CH 2 CO 2 CH 3 , C.H. 2 NHCH 2 CH 2 OCH3 , C.H. 2 NH(CH 3 )CH 2 CH 2 OCH 3 , C.H. 2 NH(CH 3 )CH 2 C(O)N(CH 3 ) 2 , C.H. 2 NH(CH 3 )CH 2 C(O)NHCH 3 , C.H. 2 CH 2 C.H.H., C.H. 2 NMe 2 , C.H. 2 NH(CH 3 )CH 2 CH 2 OH, CH 2 NH(CH 3 )CH 2 CH 2 F, C.H. 2 N + (CH 3 ) 3 , C.H. 2 NHCH 2 CHF 2 , C.H. 2 NHCH 2 CH 3 , [ka] Includes.

[0019] As used herein, the term "aryl" refers to 6~14 Monocyclic or polycyclic aromatic groups, preferably C 6~10 Monocyclic or bicyclic aromatic group or C 10~14 It refers to a polycyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Aryl refers to a C cyclic aromatic ring in which one ring is aromatic and the other rings are saturated, partially unsaturated, or aromatic. 10~14It also refers to bicyclic and tricyclic carbocycles, such as dihydronaphthyl, indenyl, indanyl or tetrahydronaphthyl (tetralinyl). Unless otherwise indicated, aryl groups may be unsubstituted or may contain one or more, and in particular, for example, halo, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, -CF 3 , -OCF 3 , -NO 2 , -CN, -NC, -OH, alkoxy, amino, -CO 2 H, -CO 2 C 1 ~C 6 Alkyl, -OCOC 1 ~C 6 Alkyl, C 3 ~C 10 Cycloalkyl, C 3 ~C 10 Heterocycloalkyl, C 5 ~C 10 Aryl and C 5 ~C 10 It may be substituted with 1 to 4 groups independently selected from heteroaryl.

[0020] The term "cycloalkyl" as used herein refers to a monocyclic or polycyclic non-aromatic carbocyclic ring, which may be fused, bridged, or spiro. A carbocyclic ring may have 3 to 10 carbon ring atoms. Possible carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclononyl.

[0021] The term "heterocycloalkyl" as used herein means a monocyclic or polycyclic (e.g., bicyclic), saturated or partially unsaturated ring system containing a total of three or more (e.g., 3-12, 4-10, 4-8, or 5-7) atoms, of which one to five (e.g., one, two, three, four, or five) 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.

[0022] Unless otherwise indicated, a cycloalkyl or heterocycloalkyl group may be unsubstituted or substituted with one or more, particularly one to four, groups. Some possible substituents include halo, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, -OCF 3 , -NO 2 , -CN, -NC, -OH, alkoxy, amino, -CO 2 H, -CO 2 C 1 ~C 6 Alkyl, -OCOC 1 ~C 6 Alkyl, C 3 ~C 10 Cycloalkyl, C 3 ~C 10 Heterocycloalkyl, C 5 ~C 10 Aryl and C 5 ~C 10 Examples of such radicals include heteroaryl.

[0023] The term "heteroaryl" as used herein refers to a monocyclic or polycyclic ring system (e.g., bicyclic) containing one to three aromatic rings and containing one to four (e.g., one, two, three, or four) heteroatoms selected from nitrogen, oxygen, and sulfur in the aromatic rings. In certain embodiments, the heteroaryl group has 5 to 20, 5 to 15, 5 to 10, or 5 to 7 atoms. Heteroaryl further includes C aryls, in which one ring is aromatic and the other ring is saturated, partially unsaturated, or aromatic. 10~14 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, benzothio, benzothiophenyl ... Examples of heteroaryl groups include phenyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, thiazolinyl, thiadiazolopyrimidyl and thienopyridyl. Unless otherwise indicated, heteroaryl groups may be unsubstituted or substituted with one or more, particularly 1 to 4 or 1 or 2, substituents. Possible substituents include halo, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, -OCF 3 , -NO 2 , -CN, -NC, -OH, alkoxy, amino, -CO 2 H, -CO 2 C 1 ~C 6 Alkyl, -OCOC 1 ~C 8 Alkyl, C3 ~C 10 Cycloalkyl, C 3 ~C 10 Heterocycloalkyl, C 5 ~C 10 Aryl and C 5 ~C 10 Examples of such radicals include heteroaryl.

[0024] As used herein, the term Boc refers to the structure [ka] Refers to...

[0025] As used herein, the term Cbz refers to the structure [ka] Refers to...

[0026] As used herein, the term Bn refers to a compound having the structure [ka] Refers to...

[0027] As used herein, the term trifluoroacetamide has the structure [ka] Refers to...

[0028] As used herein, the term trityl refers to a group having the structure [ka] Refers to...

[0029] As used herein, the term tosyl refers to the structure [ka] Refers to...

[0030] As used herein, the term Troc has the structure [ka] Refers to...

[0031] As used herein, the term Teoc has the structure [ka] Refers to...

[0032] As used herein, the term Alloc refers to the structure [ka] Refers to...

[0033] As used herein, the term Fmoc refers to the structure [ka] Refers to...

[0034] Compounds of the Disclosure The compounds disclosed herein include all pharma- ceutically acceptable isotopically labeled compounds in which one or more atoms of the compounds disclosed herein are replaced by an atom having the same atomic number, but an atomic mass or mass number different from that usually found in nature. Examples of isotopes that may be incorporated into the compounds disclosed herein include, for example, 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I and 125Radiolabeled compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as I. These radiolabeled compounds may be useful, for example, to facilitate the determination or measurement of the efficacy of a compound by identifying a site or mode of action or binding affinity to a pharmacologically important site of action. Certain isotopically labeled compounds of the present disclosure, for example those incorporating a radioisotope, are useful for studying the tissue distribution of drugs and / or substrates. The radioisotope tritium, i.e. 3 H and carbon 14, i.e. 14 C is particularly useful for this purpose in view of its ease of uptake and rapid means of detection.

[0035] Deuterium, i.e. 2 Substitution with heavier isotopes, such as H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements, and therefore may be preferable in some circumstances.

[0036] 11 C. 18 F, 15 O and 13 Substitution with positron emitting isotopes, such as N, can be useful in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those of skill in the art, or by methods analogous to those described in the Preparations and Examples below, substituting an appropriate isotopically labeled reagent for a previously used non-labeled reagent.

[0037] Isotopically labeled compounds disclosed herein can generally be prepared by conventional techniques known to those of skill in the art, or by methods analogous to those described in the accompanying Examples and Schemes, substituting an appropriate isotopically labeled reagent for a previously used non-labeled reagent.

[0038] Certain compounds disclosed herein may exist as stereoisomers (i.e., isomers that differ only in the spatial arrangement of atoms), including optical isomers and conformational isomers (or conformers). The compounds disclosed herein include all stereoisomers, both as pure individual stereoisomer preparations and enriched preparations of each, as well as racemic mixtures of such stereoisomers, as well as individual diastereomers and enantiomers that may be separated according to methods known to those skilled in the art. Additionally, the compounds disclosed herein include all tautomeric forms of those compounds.

[0039] Certain compounds disclosed herein may exist as atropisomers, which are conformational stereoisomers that arise when rotation around a single bond in a molecule is hindered or greatly slowed down as a result of steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers, both as pure individual atropisomer preparations, as concentrated preparations of each, or as unspecified mixtures of each. If the barriers to rotation around a single bond are sufficiently high and the interconversion between conformations is sufficiently slow, separation and isolation of isomeric species may be possible. For example, the following R 10 base [ka] Groups including, but not limited to, the like may exhibit restricted rotation.

[0040] Embodiment EMBODIMENT 1 In one embodiment of the present invention, the present invention provides a compound of formula (I) [ka] (In the formula, R 1 is H, halo or -CH 3 and; R 2 is H, halo or -CH 3 and; R 3 teeth, [ka] and; b is optionally a single bond or a double bond; Ring A is a monocyclic 4-7 membered ring or a bicyclic bridged, fused, or spiro 6-11 membered ring; L is a bond or NR 4 and; R 4 -H, -C 1~6 Alkyl, -C 2~6 Alkynyl, C 1~6 Alkylene-OC 1~4 Alkyl, C 1~6 Alkylene-OH, C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6 Alkylene-amide, -C(O)OH, -C(O)OC 1~4 Alkyl, -C 1~6 Alkylene-O-aryl, -N=N, -C 0~3 Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6~14 Aryl or -C 0~3 Alkylene-C 2~14 is heteroaryl; R 5 H, halo, -C 1~6 Alkyl, -C 2~6 Alkynyl, -C 1~6 Alkylene-OC 1~4 Alkyl, -C 1~6 Alkylene -OH, -C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6 Alkylene-amide, -C(O)OH, -C(O)OC 1~4 Alkyl, -C 0~6 Alkylene-OC 6-14 Aryl, -C 0~3Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6~14 Aryl, -C 0~3 Alkylene-C 2~14 heteroaryl or cyano; R 5a -H, -C 1~6 Alkyl, -C 2~6 Alkynyl, -C 1~6 Alkylene-OC 1~4 Alkyl, -C 1~6 Alkylene -OH, -C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6 Alkylene-amide, -C(O)OH, -C(O)OC 1~4 Alkyl, -C 0~6 Alkylene-OC 6~14 Aryl, -C 0~3 Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6~14 Aryl or -C 0~3 Alkylene-C 2~14 heteroaryl; R 5b -H, -C 1~6 Alkyl, -C 2~6 Alkynyl, -C 1~6 Alkylene-OC 1~4 Alkyl, -C 1~6 Alkylene -OH, -C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6Alkylene-amide, -C(O)OH, -C(O)OC 1~4 Alkyl, -C 0~6 Alkylene-OC 6~14 Aryl, -C 0~3 Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6~14 Aryl or -C 0~3 Alkylene-C 2~14 heteroaryl; or R 5a and R 5b together can represent =O or =N=N; R 6 H, halo, -C 1~6 Alkyl, -C 2~6 Alkynyl, -C 1~6 Alkylene-OC 1~4 Alkyl, -C 1~6 Alkylene -OH, -C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6 Alkylene-amide, -C(O)OH, -C(O)OC 1~4 Alkyl, -C 0~6 Alkylene-OC 6-14 Aryl, -C 0~3 Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6~14 Aryl or -C 0~3 Alkylene-C 2~14 is heteroaryl; R 5a and R 6amay, together with the atoms to which they are attached, form a 3- to 6-membered ring, optionally containing 1 or 2 heteroatoms selected from O, S or N; or R 5a and R 6a is absent when b is a double bond; R 6a is H or -C 1~6 is alkyl; R 6b -H, -C 1~6 Alkyl, -C 2~6 Alkynyl, C 1~6 Alkylene-OC 1~4 Alkyl, C 1~6 Alkylene-OH, C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6 Alkylene-amide, -C(O)OH, -C(O)OC 1~4 Alkyl, -C 1~6 Alkylene-O-aryl, -C 0~3 Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6~14 Aryl, -C 0~3 Alkylene-C 2~14 is heteroaryl or cyclo; or R 6a and R 6b taken together can represent =O; R 7 is H or C 1~8 is alkyl; R 8 H, OH, NR a R b and Here, R a and R b are each independently H, halo, -C 1~6 Alkyl, -C 2~6 is alkynyl; Here, ring A or R 4 , R 5 , R 5a , R 5b , R 6 , R 6a , R 6b , R 7 and R 8 Either C 1~6 Alkyl, -C 2~6 Alkynyl, -C 1~6 Alkylene-OC 1~4 Alkyl, -C 1~6 Alkylene -OH, -C 1~6 Haloalkyl, -C 1~6 Alkyleneamines, -C 0~6 Alkylene-amide, -C(O)OC 1~4 Alkyl, -C 1~6 Alkylene-O-aryl, -C 0~3 Alkylene-C(O)C 1~4 Alkylene-OH, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -C 0~3 Alkylene-C 3~14 Cycloalkyl, -C 0~3 Alkylene-C 2~14 Heterocycloalkyl, -C 0~3 Alkylene-C 6-14 Aryl or -C 0~3 Alkylene-C 2~14 Heteroaryl groups may be unsubstituted or substituted, where permitted, with halo, -C 1~6 Alkyl, -OC 1~6 Alkyl, -OH or -C 1~6 and optionally substituted with 1, 2, 3, or 4 substituents independently selected from alkyl-CN. A compound having the structure or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0041] EMBODIMENT 2 In a second embodiment, the present invention provides a compound having the structure: [ka] The compound of embodiment 1 includes compounds having the formula:

[0042] EMBODIMENT 3 The compound of any one of embodiments 1-2, wherein ring A is a monocyclic 4- to 7-membered ring.

[0043] EMBODIMENT 4 The compound of any one of embodiments 1-2, wherein Ring A is a monocyclic ring or a bicyclic ring.

[0044] EMBODIMENT 5 The compound of any one of embodiments 1-2, wherein Ring A is a spiro 6-11 membered ring.

[0045] EMBODIMENT 6 R 1 The compound of any one of embodiments 1-5, wherein is F.

[0046] EMBODIMENT 7 R 2 The compound of any one of embodiments 1-6, wherein is Cl.

[0047] EMBODIMENT 8 R 5 H, halo, -C 1~6 Alkyl or -C 0~3 Alkylene-C 6~14 The compound of any one of embodiments 1-7, wherein is aryl.

[0048] EMBODIMENT 9 R 5 is H.

[0049] EMBODIMENT 10 R 5 The compound of embodiment 8, wherein is halo.

[0050] EMBODIMENT 11 R 5 The compound of embodiment 8, wherein is Cl or F.

[0051] EMBODIMENT 12 R 5 -C 1~6 The compound of embodiment 8, wherein the is alkyl.

[0052] EMBODIMENT 13 R 5 -C 0~3 Alkylene C 6~14 The compound of embodiment 8, wherein the is aryl.

[0053] EMBODIMENT 14 R 5a is H, -C, unless b is a double bond. 1~6 Alkyl, -C 1~6 Alkylene -OH, -C(O)OH, -C(O)OC 1~4 Alkyl and -C 0~3 Alkylene-C 2~14 The compound of any one of embodiments 1-13, wherein the heterocycloalkyl is selected from:

[0054] EMBODIMENT 15 R 5a is H.

[0055] EMBODIMENT 16 R 5a -C 1~6 The compound of embodiment 14, wherein the is alkyl.

[0056] EMBODIMENT 17 R 5a -C 1~6 The compound of embodiment 14, which is alkylene-OH.

[0057] EMBODIMENT 18 R 5a The compound of embodiment 14, wherein is —C(O)OH.

[0058] EMBODIMENT 19 R 5a is -C(O)OC 1~4 The compound of embodiment 14, wherein the is alkyl.

[0059] EMBODIMENT 20 R 5a -C 0~3 Alkylene-C 2~14 The compound of embodiment 14, which is heterocycloalkyl.

[0060] EMBODIMENT 21 R 6 is R 3 The following structure [ka] Unless it has H, halo or -C 1~6 The compound of any one of embodiments 1-20, wherein is alkyl.

[0061] EMBODIMENT 22 R 6 The compound of embodiment 21, wherein

[0062] EMBODIMENT 23 R 6 22. The compound of embodiment 21, wherein is halo.

[0063] EMBODIMENT 24 R 6 -C 1~6 22. The compound of embodiment 21, wherein the is alkyl.

[0064] EMBODIMENT 25 R 6b is R 3 The following structure [ka] H, -C, unless 1~6 Alkyl, C 1~6 Alkylene -OH, -C(O)OH, -C(O)OC 1~4 Alkyl or -C 0~3 Alkylene-C 2~14 The compound of any one of embodiments 1-24, which is heterocycloalkyl.

[0065] EMBODIMENT 26 R 6bThe compound of embodiment 25, wherein

[0066] EMBODIMENT 27 R 6b -C 1~6 The compound of embodiment 25, wherein the is alkyl.

[0067] EMBODIMENT 28 R 6b is C 1~6 The compound of embodiment 25, which is alkylene-OH.

[0068] EMBODIMENT 29 R 6b The compound of embodiment 25, wherein is -C(O)OH.

[0069] EMBODIMENT 30 R 6b is -C(O)OC 1~4 The compound of embodiment 25, wherein the is alkyl.

[0070] EMBODIMENT 31 R 6b -C 0~3 Alkylene-C 2~14 The compound of embodiment 25, which is heterocycloalkyl.

[0071] EMBODIMENT 32 R 6a and R 6b is R 3 The following structure [ka] The compound of any one of embodiments 1-24, unless having the formula:

[0072] EMBODIMENT 33 R 6a is R 3 but The following structure [ka] and b is not a double bond, H or -C1~6 The compound of any one of embodiments 1-31, wherein is alkyl.

[0073] EMBODIMENT 34 R 6a is H.

[0074] EMBODIMENT 35 R 6a -C 1~6 The compound of embodiment 33, wherein the is alkyl.

[0075] EMBODIMENT 36 The compound of any one of embodiments 1-35, wherein L is a bond.

[0076] EMBODIMENT 37 L is NR 4 The compound of any one of embodiments 1-35, wherein

[0077] EMBODIMENT 38 R 4 is H or -C 1~6 The compound of embodiment 37, wherein the is alkyl.

[0078] EMBODIMENT 39 R 4 is H.

[0079] EMBODIMENT 40 R 4 -C 1~6 The compound of embodiment 37, wherein the is alkyl.

[0080] EMBODIMENT 41 The compound of any one of embodiments 1-40, wherein b is a single bond.

[0081] EMBODIMENT 42 The compound of any one of embodiments 1-40, wherein b is a double bond, unless otherwise specified.

[0082] EMBODIMENT 43 R 3 teeth, [ka] The compound of any one of embodiments 1-35, wherein

[0083] EMBODIMENT 44 R 5a and R 6a may form, together with the atom to which they are attached, a 3-6 membered ring, optionally containing 1 or 2 heteroatoms selected from O, S, or N.

[0084] EMBODIMENT 45 R 3 teeth, [ka] The compound of any one of embodiments 1-35, wherein

[0085] EMBODIMENT 46 R 3 teeth, [ka] The compound of any one of embodiments 1-20, wherein

[0086] EMBODIMENT 47 R 5b -H, -C 1~6 Alkyl, -C 1~6 Alkylene -OH, -C(O)OH, -C(O)OC 1~4 Alkyl or C 0~3 Alkylene-C 2~14 The compound of embodiment 46, wherein the heterocycloalkyl is selected from:

[0087] EMBODIMENT 48 R 5b The compound of embodiment 47, wherein

[0088] EMBODIMENT 49 R 5b -C 1~6The compound of embodiment 47, wherein the is alkyl.

[0089] EMBODIMENT 50 R 5b -C 1~6 The compound of embodiment 47, which is alkylene-OH.

[0090] EMBODIMENT 51 R 5b The compound of embodiment 47, wherein is —C(O)OH.

[0091] EMBODIMENT 52 R 5b is -C(O)OC 1~4 The compound of embodiment 47, wherein the is alkyl.

[0092] EMBODIMENT 53 R 5b -C 0~3 Alkylene-C 2~14 The compound of embodiment 47, which is heterocycloalkyl.

[0093] EMBODIMENT 54 R 5a and R 5b The compound of embodiment 46, wherein together may represent =O.

[0094] EMBODIMENT 55 R 5a and R 5b may together represent =N=N.

[0095] EMBODIMENT 56 R 3 teeth, [ka] The compound of any one of embodiments 1-35, wherein

[0096] EMBODIMENT 57 R 5a and R 6amay, together with the atom to which they are attached, form a 3-6 membered ring, optionally containing 1 or 2 heteroatoms selected from O, S, or N.

[0097] EMBODIMENT 58 R 3 teeth, [ka] The compound of any one of embodiments 1-35, wherein

[0098] EMBODIMENT 59 R 3 teeth, [ka] The compound of any one of embodiments 1-40, wherein

[0099] EMBODIMENT 60 R 5a and R 6a may, together with the atom to which they are attached, form a 3-6 membered ring, optionally containing 1 or 2 heteroatoms selected from O, S, or N.

[0100] EMBODIMENT 61 R 3 teeth, [ka] The compound of any one of embodiments 1-40, wherein

[0101] EMBODIMENT 62 R 3 teeth, [ka] The compound of any one of embodiments 1-35, wherein

[0102] EMBODIMENT 63 R 5a and R 6amay, together with the atom to which they are attached, form a 3-6 membered ring, optionally containing 1 or 2 heteroatoms selected from O, S, or N.

[0103] EMBODIMENT 64 R 3 teeth, [ka] The compound of any one of embodiments 1-35, wherein

[0104] EMBODIMENT 65 R 7 is H or C 1~8 The compound of any one of embodiments 62-64, wherein is alkyl.

[0105] EMBODIMENT 66 R 7 is H.

[0106] EMBODIMENT 67 R 7 -C 1~8 The compound of embodiment 65, wherein the is alkyl.

[0107] EMBODIMENT 68 R 3 teeth, [ka] [ka] The compound of any one of embodiments 1-2, selected from:

[0108] EMBODIMENT 69 R 1 The compound of embodiment 68, wherein is F.

[0109] EMBODIMENT 70 R 2 The compound of embodiment 68 or 69, wherein is Cl.

[0110] EMBODIMENT 71 R 8 The compound of any one of embodiments 1-70, wherein

[0111] EMBODIMENT 72 R 8 The compound of any one of embodiments 1-70, wherein is OH.

[0112] EMBODIMENT 73 R 8 is NR a R b The compound of any one of embodiments 1-70, wherein

[0113] EMBODIMENT 74 NR a R b NH 2 74. The compound of embodiment 73, wherein

[0114] EMBODIMENT 75 the below described: [ka] [ka] [ka] [ka] [ka] [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0115] EMBODIMENT 76 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0116] EMBODIMENT 77 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0117] EMBODIMENT 78 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0118] EMBODIMENT 79 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0119] Embodiment 80 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0120] Embodiment 81 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0121] EMBODIMENT 82 the below described: [ka] or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of the stereoisomer, or a pharma- ceutically acceptable salt of the atropisomer.

[0122] EMBODIMENT 83 The compound of any one of embodiments 1-80, in the form of a pharma- ceutically acceptable salt.

[0123] EMBODIMENT 84 A pharmaceutical composition comprising a compound of any one of embodiments 1-83 and a pharma- ceutically acceptable excipient.

[0124] EMBODIMENT 85 A kit for treating cancer, comprising a compound of any one of embodiments 1-84 and an additional pharma- ceutical active compound.

[0125] EMBODIMENT 86 The kit of embodiment 85, wherein the cancer is a solid tumor.

[0126] EMBODIMENT 87 The kit of embodiment 85, wherein the cancer is non-small cell lung cancer.

[0127] EMBODIMENT 88 The kit of embodiment 85, wherein the cancer is colorectal cancer.

[0128] EMBODIMENT 89 The kit of embodiment 85, wherein the cancer is pancreatic cancer.

[0129] EMBODIMENT 90 86. The kit of embodiment 85, wherein the cancer is a KRAS G12C mutant cancer.

[0130] EMBODIMENT 91 86. The kit of embodiment 85, wherein the cancer is a KRAS G12D mutant cancer.

[0131] EMBODIMENT 92 A method of inhibiting KRAS G12C in a cell, comprising contacting the cell with a compound of any one of embodiments 1-91.

[0132] EMBODIMENT 93 A method of inhibiting KRAS G12D in a cell, comprising contacting the cell with a compound of any one of embodiments 1-91.

[0133] EMBODIMENT 94 A method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a compound of any one of embodiments 1-84.

[0134] EMBODIMENT 95 95. The method of embodiment 94, wherein the cancer is a solid tumor cancer.

[0135] EMBODIMENT 96 95. The method of embodiment 94, wherein the cancer is lung cancer, pancreatic cancer, endometrial cancer, appendix cancer, small intestine cancer or colorectal cancer.

[0136] EMBODIMENT 97 97. The method of embodiment 96, wherein the cancer is lung cancer.

[0137] EMBODIMENT 98 97. The method of embodiment 96, wherein the lung cancer is non-small cell lung cancer.

[0138] EMBODIMENT 99 97. The method of embodiment 96, wherein the cancer is pancreatic cancer.

[0139] EMBODIMENT 100 97. The method of embodiment 96, wherein the cancer is colorectal cancer.

[0140] EMBODIMENT 101 97. The method of embodiment 96, wherein the cancer is endometrial cancer.

[0141] EMBODIMENT 102 97. The method of embodiment 96, wherein the cancer is appendiceal cancer.

[0142] EMBODIMENT 103 97. The method of embodiment 96, wherein the cancer is small intestine cancer.

[0143] EMBODIMENT 104 95. The method of embodiment 94, further comprising administering to a patient in need thereof a therapeutically effective amount of an additional pharma- ceutical active compound.

[0144] EMBODIMENT 105 The method of embodiment 104, wherein the compound of any one of embodiments 1-84 is administered prior to the additional pharma- ceutically active compound.

[0145] EMBODIMENT 106 The method of embodiment 96, wherein the compound of any one of embodiments 1-84 is administered simultaneously with an additional pharma- ceutically active compound.

[0146] EMBODIMENT 107 The method of embodiment 96, wherein the compound of any one of embodiments 1-84 is administered after the additional pharma- ceutically active compound.

[0147] EMBODIMENT 108 The method of embodiment 104, wherein the additional pharma- ceutical active compound is an anti-PD-1 antagonist.

[0148] EMBODIMENT 109 The method of embodiment 108, wherein the anti-PD-1 antagonist is nivolumab.

[0149] Embodiment 110 The method of embodiment 108, wherein the anti-PD-1 antagonist is pembrolizumab.

[0150] EMBODIMENT 111 The method of embodiment 108, wherein the anti-PD-1 antagonist is AMG 404.

[0151] EMBODIMENT 112 The method of embodiment 104, wherein the additional pharma- ceutical active compound is a MEK inhibitor.

[0152] EMBODIMENT 113 The method of embodiment 112, wherein the MEK inhibitor is trametinib.

[0153] EMBODIMENT 114 The method of embodiment 104, wherein the additional pharma- ceutical active compound is an SHP2 inhibitor.

[0154] EMBODIMENT 115 The method of embodiment 114, wherein the SHP2 inhibitor is RMC-4630.

[0155] EMBODIMENT 116 Use of a compound of any one of embodiments 1-84 for treating cancer in a subject.

[0156] EMBODIMENT 117 A compound of any one of embodiments 1-84 in the preparation of a medicament for treating cancer.

[0157] EMBODIMENT 118 The compound of embodiment 116, wherein the cancer is a solid tumor.

[0158] EMBODIMENT 119 A compound of any one of embodiments 1 to 83 or a pharmaceutical composition of embodiment 84 for use as a medicament.

[0159] Contemplated halogenating agents include, but are not limited to, chlorine, bromine, N-chlorosuccinimide, and N-bromosuccinimide, optionally in the presence of a catalyst such as iron or aluminum. A synthetic chemist of ordinary skill will readily appreciate that other halogenating agents and catalysts can be used.

[0160] Possible amidating agents include, but are not limited to, N,N'-diisopropylcarbodiimide, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide, benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, thionyl chloride, isobutyl chloroformate, diethyl cyanophosphonate, carbonyldiimidazole and polyphosphonic anhydride. A synthetic chemist of ordinary skill will readily understand that other amidating agents can be used.

[0161] Contemplated sulfurizing agents include, but are not limited to, sulfur, phosphorus pentasulfide, and Lawesson's reagent. A synthetic chemist of ordinary skill will readily appreciate that other sulfurizing agents can be used.

[0162] Possible oxidizing agents include, but are not limited to, hydrogen peroxide, iodobenzene diacetate, t-butyl hydroperoxide, N-bromosuccinimide, and ammonium peroxodisulfate. A synthetic chemist of ordinary skill will readily appreciate that other oxidizing agents can be used.

[0163] Possible metallation agents include, but are not limited to, bis(pinacolato)diboron, magnesium, zinc, hexamethyldistannane, and n-butyllithium. A synthetic chemist of ordinary skill will readily appreciate that other metallation agents and catalysts can be used.

[0164] Possible activating agents include, but are not limited to, sodium nitrite and t-butyl nitrite. A synthetic chemist of ordinary skill will readily appreciate that other activating agents can be used.

[0165] Possible cross-coupling reactions include, but are not limited to, Suzuki coupling, Negishi coupling, Hiyama coupling, Kumada coupling and Stille coupling. A chemist of ordinary skill will readily appreciate that the couplings shown in the methods below can be carried out under a number of conditions.

[0166] Pharmaceutical Compositions, Administration and Routes of Administration Also provided herein are pharmaceutical compositions comprising the compounds disclosed herein, together with a pharma- ceutically acceptable excipient, such as a diluent or carrier. Compounds and pharmaceutical compositions suitable for use in the present invention include those in which the compound can be administered in an amount effective to achieve its intended purpose. Administration of the compounds is described in more detail below.

[0167] Suitable pharmaceutical formulations can be determined by those skilled in the art depending on the route of administration and the desired dosage. For example, see Remington's Pharmaceutical Sciences, 1435-712 (18th ed., Mack Publishing Co, Easton, Pennsylvania, 1990). Formulations can affect the physical state, stability, in vivo release rate and in vivo excretion rate of the administered drug. Depending on the route of administration, suitable doses can be calculated according to body weight, body surface area or organ size. Further refinement of calculations required to determine appropriate therapeutic doses can be routinely performed by those skilled in the art without undue experimentation, especially in light of the dosage information and assays disclosed herein, and pharmacokinetic data obtained in animal or human clinical trials.

[0168] The phrase "pharmacologically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not produce adverse allergic or other untoward reactions when administered to animals or humans. As used herein, "pharmacologically acceptable excipients" include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such excipients for pharma- ceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the therapeutic composition, its use in the therapeutic composition is contemplated. Supplementary active ingredients can also be incorporated into the composition. In an exemplary embodiment, the formulation contains corn syrup solids, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tricalcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dipotassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, calcium carbonate, L-glutamic acid, L-cystine dihydrate. Hydrochloride, L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, mixed tocopherols, calcium pantothenate, copper sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0169] The compound may be present in the pharmaceutical composition as a pharma- ceutically acceptable salt. As used herein, "pharmaceutically acceptable salt" includes, for example, base addition salts and acid addition salts.

[0170] 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 compounds can also be prepared with pharma-ceutically acceptable cations. Suitable pharma-ceutically acceptable cations are well known to those skilled in the art and include alkali, alkaline earth, ammonium and quaternary ammonium cations. Carbonates or bicarbonates are also possible. Examples of metals used as cations include sodium, potassium, magnesium, ammonium, calcium or iron. Examples of suitable amines include isopropylamine, trimethylamine, histidine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine and procaine.

[0171] Pharmaceutically acceptable acid addition salts include salts of inorganic or organic acids. Examples of suitable acid salts include hydrochloride, formate, acetate, citrate, salicylate, nitrate, and phosphate. Other suitable pharma- ceutically acceptable salts are well known to those skilled in the art, and include, for example, formate, acetic acid, citric acid, oxalic acid, tartaric acid, or mandelic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, or phosphoric acid; organic carboxylic acid, sulfonic acid, sulfoacid, or phosphoric acid, or N-substituted sulfamic acid, such as acetic acid, trifluoroacetic acid (TFA), propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, and with the 20 alpha-amino acids involved in the synthesis of natural proteins, such as glutamic acid or aspartic acid, and also with phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane 1,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene 2-sulfonic acid, naphthalene 1,5-disulfonic acid, 2- or 3-phosphoglyceric acid, glucose 6-phosphate, N-cyclohexylsulfamic acid (with the formation of cyclamate) or with other acid organic compounds, such as ascorbic acid.

[0172] Pharmaceutical compositions containing the compounds disclosed herein can be manufactured in a conventional manner, for example by conventional mixing, dissolving, granulating, dragee-making, pulverizing, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation depends on the route of administration chosen.

[0173] For oral administration, suitable compositions can be easily formulated by combining the compounds disclosed herein with pharma- ceutically acceptable excipients, such as carriers well known in the art. Such excipients and carriers allow the compounds of the present invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by the patient to be treated. Pharmaceutical preparations for oral use can be obtained by adding the compounds disclosed herein with solid excipients, optionally grinding the resulting mixture, and processing the granulated mixture, after adding suitable auxiliaries as necessary, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. Disintegrants can be added as necessary. Pharmaceutically acceptable ingredients are well known for various types of formulations and can be, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavoring agents, coatings, preservatives, dyes, thickeners, adjuvants, antimicrobial agents, antioxidants, and carriers for various formulation types.

[0174] When a therapeutically effective amount of a compound disclosed herein is administered orally, the composition is typically in the form of a solid (e.g., a tablet, capsule, pill, powder or lozenge) or a liquid preparation (e.g., an aqueous suspension, solution, elixir or syrup).

[0175] When administered in tablet form, the composition may further contain a functional solid, such as gelatin or an adjuvant, and / or a functional solid carrier. The tablet, capsule, and powder may contain about 1 to about 95% of the compound, and preferably about 15 to about 90% of the compound.

[0176] When administered in the form of a solution or suspension, a functional liquid and / or a functional liquid carrier, such as water, petroleum, or oil of animal or plant origin, can be added. The liquid form of the composition can further include a saline solution, a sugar alcohol solution, a dextrose or other sugar solution, or a glycol. When administered in the form of a solution or suspension, the composition can contain about 0.5 to about 90% by weight of the compound disclosed herein, preferably about 1 to about 50% of the compound disclosed herein. In one contemplated embodiment, the liquid carrier is non-aqueous or substantially non-aqueous. For administration in liquid form, the composition can be provided as a rapidly dissolving solid formulation that is dissolved or suspended immediately prior to administration.

[0177] When a therapeutically effective amount of the compounds disclosed herein is administered by intravenous, cutaneous or subcutaneous injection, the composition is in the form of a pyrogen-free parenterally acceptable aqueous solution. Preparation of such parenterally acceptable solutions, with due consideration of pH, isotonicity, stability, etc., is within the skill of the art. A preferred composition for intravenous, cutaneous or subcutaneous injection typically contains an isotonic vehicle in addition to the compounds disclosed herein. Such compositions can be prepared for administration as a solution of a free base or a pharmacologically acceptable salt in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof, as well as in oils. Under ordinary conditions of storage and use, these preparations can optionally contain a preservative to prevent the growth of microorganisms.

[0178] Injectable compositions can include sterile aqueous solutions, suspensions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions, suspensions or dispersions. In all embodiments, the form must be sterile and fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must resist the contaminating action of microorganisms such as bacteria and fungi, optionally by including preservatives. The carrier can be, for example, a solvent or dispersion medium containing water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. In one possible embodiment, the carrier is non-aqueous or substantially non-aqueous. The proper fluidity can be maintained, for example, by the use of a coating agent such as lecithin, by maintaining the particle size of the compound required in the embodiment of the dispersion, and by the use of surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many embodiments, it is preferable to include an isotonic agent, for example, sugar or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0179] Sterile injectable solutions are prepared by incorporating the active compound in the required amount in a suitable solvent together with various other necessary ingredients listed above, followed by filtration sterilization.Generally, dispersion is prepared by incorporating various sterilized active ingredients into a sterile vehicle that contains basic dispersion medium and other desired ingredients from the ingredients listed above.In the embodiment of sterile powder for preparing sterile injectable solutions, the preferred preparation method is vacuum drying and freeze-drying technology, which obtains powder of active ingredient plus any additional desired ingredients from the solution that has been previously sterilized and filtered.

[0180] Slow or sustained release formulations can also be prepared to provide a controlled release of active compounds in contact with body fluids in the digestive tract and provide a substantially constant and effective level of active compounds in plasma. For example, release can be controlled by one or more of dissolution, diffusion and ion exchange. Furthermore, the slow release approach can enhance absorption via saturable or restricted pathways in the digestive tract. For example, the compound can be embedded in a polymer matrix of a biodegradable polymer, a water-soluble polymer or a mixture of both, and optionally a suitable surfactant, for this purpose. Embedding in this context can mean incorporating microparticles in a polymer matrix. Controlled release formulations can also be obtained by encapsulation of dispersed microparticles or emulsified microdroplets via known dispersion or emulsion coating techniques.

[0181] For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of aerosol spray dispensed from pressurized packs or nebulizers using a suitable propellant.In pressurized aerosol embodiments, the dosage unit can be determined by providing a valve to deliver a metered amount.Capsules and cartridges, for example of gelatin, for use in an inhaler or insufflator can be formulated to contain a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0182] The compounds disclosed herein can be formulated for parenteral administration by injection (e.g., by bolus injection or continuous infusion).The formulation for injection can be provided in unit dosage form (e.g., in ampoules or multi-dose containers) with added preservatives.The composition can take the form of a suspension, solution or emulsion in an oily or aqueous vehicle, and can contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0183] Pharmaceutical preparations for parenteral administration include aqueous solutions of the compound in water-soluble form. Additionally, suspensions of the compound can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound and allow for the preparation of highly concentrated solutions. Alternatively, the compositions of the present invention can be in powder form for constitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0184] The compounds disclosed herein can also be formulated into rectal compositions such as suppositories or retention enemas (e.g., containing conventional suppository bases). In addition to the formulations described above, the compounds can also be formulated as depot preparations. Such long-acting formulations can be administered by injection (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or can be formulated as sparingly soluble derivatives, e.g., as a sparingly soluble salt.

[0185] In particular, the compounds disclosed herein may be administered orally, bucally or sublingually in the form of tablets containing excipients such as starch or lactose, or in capsules or ovoids, alone or in mixture with excipients, or in the form of elixirs or suspensions containing flavorings or colorings. Such liquid preparations may be prepared using pharma- ceutically acceptable additives such as suspending agents. The compounds may also be injected parenterally, for example, intravenously, intramuscularly, subcutaneously or intracoronarily. For parenteral administration, the compounds are most often used in the form of a sterile aqueous solution, which may contain other substances, for example, salts or sugar alcohols such as mannitol or glucose, to make the solution isotonic with blood.

[0186] For veterinary use, the compounds disclosed herein are administered in an appropriately acceptable formulation in accordance with normal veterinary practice, and a veterinarian can readily determine the most appropriate dosing regimen and route of administration for a particular animal.

[0187] In some embodiments, all components required for the treatment of KRAS-related disorders, using the compounds disclosed herein alone or in combination with another drug or intervention conventionally used for the treatment of such diseases, can be packaged into a kit.Specifically, the present invention provides a kit that includes a set of packaged pharmaceuticals for use in the treatment and intervention of a disease, comprising the compounds disclosed herein, and buffers and other components for preparing a deliverable form of said pharmaceuticals, and / or a device for delivering such pharmaceuticals, and / or any agent used together with the compounds disclosed herein in combination therapy, and / or instructions for treating the disease packaged together with the pharmaceuticals.The instructions can be fixed on any tangible medium, such as printed paper or computer-readable magnetic or optical media, or can be instructions that refer to a remote computer data source, such as a World Wide Web page accessible via the Internet.

[0188] "Therapeutically effective amount" refers to an amount effective to treat, prevent progression, or alleviate existing symptoms of the subject being treated. Determining an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. In general, "therapeutically effective dose" refers to the amount of a compound that produces a desired effect. For example, in a preferred embodiment, a therapeutically effective amount of a compound disclosed herein reduces KRAS activity by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90% compared to a control.

[0189] The amount of compound administered may depend on the subject being treated, the subject's age, health, sex and weight, type of combination treatment (if any), severity of the affliction, the nature of the desired effect, the mode and frequency of treatment, and the judgment of the prescribing physician. The frequency of administration may also depend on the pharmacodynamic effect on arterial oxygen pressure. However, the most preferred dosage can be adjusted for each individual subject, as can be understood and determined by those skilled in the art without undue experimentation. This usually involves adjusting the standard dosage (e.g., reducing the dosage if the patient is underweight).

[0190] While individual needs vary, determining the optimal range of effective amounts of the compounds is within the skill of the art. For administration to humans in the curative or prophylactic treatment of conditions and disorders identified herein, for example, a typical dosage of the compounds of the invention may be 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, about 2.5 mg / day (0.5 mg / kg x 5 kg) to about 5000 mg / day (50 mg / kg x 100 kg). For example, the dosage of the compound may be from about 0.1 mg / kg / day to about 50 mg / kg / day, from about 0.05 mg / kg / day to about 10 mg / kg / day, from about 0.05 mg / kg / day to about 5 mg / kg / day, from about 0.05 mg / kg / day to about 3 mg / kg / day, from about 0.07 mg / kg / day to about 3 mg / kg / day, from about 0.09 mg / kg / day to about 3 mg / kg / day, from about 0.05 mg / kg / day to about 0.1 mg / kg / day The dose may be about 0.1 mg / kg / day to about 1 mg / kg / day, about 1 mg / kg / day to about 10 mg / kg / day, about 1 mg / kg / day to about 5 mg / kg / day, about 1 mg / kg / day to about 3 mg / kg / day, about 3 mg / day to about 1000 mg / day, about 5 mg / day to about 500 mg / day, about 10 mg / day to about 200 mg / day, about 3 mg / day to about 100 mg / day, or about 100 mg / day to about 250 mg / day. Such doses may be administered in a single dose or may be divided into multiple doses.

[0191] How to Use KRAS G12C Inhibitors The present disclosure provides a method for inhibiting RAS-mediated cell signaling, comprising contacting a cell with an effective amount of one or more compounds disclosed herein. Inhibition of RAS-mediated signaling can be assessed and demonstrated by a variety of methods known in the art. Non-limiting examples include: (a) a decrease in the GTPase activity of RAS; (b) a decrease in GTP binding affinity or an increase in GDP binding affinity; (c) an increase in Koff of GTP or a decrease in Koff of GDP; (d) a decrease in the level of downstream signaling molecules of the RAS pathway, such as a decrease in the level of pMEK, pERK or pAKT; and / or (e) a decrease in the binding of the RAS complex to downstream signaling molecules, including but not limited to Raf. Kits and commercially available assays can be utilized to determine one or more of the above.

[0192] The present disclosure also provides methods of using a compound or pharmaceutical composition of the present disclosure to treat a disease state, including, but not limited to, a condition caused by a G12C KRAS, G12D KRAS, HRAS, or NRAS mutation (e.g., cancer).

[0193] In some embodiments, a method of treating cancer is provided, comprising administering to a subject in need thereof any of the aforementioned pharmaceutical compositions comprising an effective amount of a compound disclosed herein. In some embodiments, the cancer is mediated by KRAS, HRAS, or NRAS G12C or G12D mutations. 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.

[0194] In some embodiments, the disclosure provides a method of treating a disorder in a subject in need thereof, comprising determining whether the subject has a KRAS, HRAS or NRAS G12C or G12D mutation, and if the subject is determined to have a KRAS, HRAS or NRAS G12C or G12D mutation, administering to the subject a therapeutically effective dose of at least one compound disclosed herein, or a pharma- ceutical acceptable salt thereof.

[0195] The compounds disclosed herein have the potential to inhibit anchorage-independent cell proliferation and thus inhibit tumor metastasis. Thus, in another embodiment, the present disclosure provides a method of inhibiting tumor metastasis, comprising administering an effective amount of a compound disclosed herein.

[0196] KRAS, HRAS or NRAS G12C or G12D mutations have also been identified in hematological malignancies (e.g., cancers affecting the blood, bone marrow and / or lymph nodes). Accordingly, certain embodiments are directed to the administration of the compounds disclosed herein (e.g., in the form of a pharmaceutical composition) to patients in need of treatment for hematological malignancies. Such malignancies 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 myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia (CML), acute monocytic leukemia (AMoL) and / or other leukemias. In other embodiments, the compounds are useful for treating lymphomas, such as all subtypes of Hodgkin's lymphoma or non-Hodgkin's lymphoma. In various embodiments, the compounds are useful in the treatment of plasma cell malignancies such as multiple myeloma, mantle cell lymphoma, and Waldenstrom's macroglobulinemia.

[0197] Determining whether a tumor or cancer contains a G12C KRAS, HRAS or NRAS mutation or a G12D KRAS, HRAS or NRAS mutation can be made by evaluating the nucleotide sequence encoding the KRAS, HRAS or NRAS protein, by evaluating the amino acid sequence of the KRAS, HRAS or NRAS protein, or by evaluating the characteristics of a putative KRAS, HRAS or NRAS mutant protein. Wild-type human KRAS, HRAS or NRAS sequences are known in the art (e.g., Accession No. NP203524).

[0198] Methods for detecting mutations in the nucleotide sequence of KRAS, HRAS or NRAS are well known to those skilled in the art. These methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) assay, real-time PCR assay, PCR sequencing, mutant allele-specific PCR amplification (MASA) assay, direct sequencing, primer extension reaction, electrophoresis, oligonucleotide ligation assay, hybridization assay, TaqMan assay, SNP genotyping assay, high resolution melting assay and microarray analysis. In some embodiments, samples are evaluated for G12C or G12D KRAS, HRAS or NRAS mutations by real-time PCR. In real-time PCR, a fluorescent probe specific for KRAS, HRAS or NRAS G12C or G12D mutations is used. If a mutation is present, the probe binds and fluorescence is detected. In some embodiments, KRAS, HRAS or NRAS G12C or G12D mutations are identified using direct sequencing of specific regions in the KRAS, HRAS or NRAS genes (e.g., exon 2 and / or exon 3). This technique identifies all possible mutations in the sequenced region.

[0199] Methods for detecting mutations in KRAS, HRAS or NRAS proteins are known to those of skill in the art, including, but not limited to, detection of KRAS, HRAS or NRAS mutations using binding agents (e.g., antibodies) specific for the mutant protein, protein electrophoresis and Western blots, and direct peptide sequencing.

[0200] A variety of samples can be used in the method of determining whether a tumor or cancer contains a G12C or G12D KRAS, HRAS or NRAS mutation. In some embodiments, the sample is taken from a subject having a tumor or cancer. In some embodiments, the sample is a fresh tumor / cancer sample. In some embodiments, the sample is a frozen tumor / cancer sample. In some embodiments, the sample is a formalin-fixed paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

[0201] The present disclosure also relates to a method of treating a hyperproliferative disorder in a mammal, comprising administering to the mammal a therapeutically effective amount of a compound disclosed herein or a pharma- ceutically acceptable salt thereof. In some embodiments, the method is for treating a hyperproliferative disorder in a mammal, including solid tumor cancer, acute myeloid leukemia, adolescent cancer, childhood adrenocortical carcinoma, AIDS-related cancer (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical teratoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoma, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, small intestine cancer, or ovarian cancer. childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), germinoma, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer cancer), gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, cardiac cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, recurrent metastatic cervical squamous cell carcinoma of unknown primary, midline carcinoma, oral cancer cnacer), multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma and osteosarcoma of bone, nasal and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancerThe present invention relates to the treatment of subjects suffering from cancer, such as pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach (gastric) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, childhood anomaly cancer, urachal cancer, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer. In some embodiments, the method relates to the treatment of non-cancerous hyperproliferative disorders, such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hyperplasia (BPH)).

[0202] In some embodiments, the method for treating is directed to treating lung cancer, the method comprising administering to a subject in need thereof an effective amount of any of the compounds described above (or pharmaceutical compositions comprising the compounds). 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 lung cancers treatable by the compounds disclosed herein include, but are not limited to, ductal tumors, carcinoid tumors, and undifferentiated carcinomas.

[0203] The present disclosure further provides a method of modulating the activity of a G12C and / or G12D mutant KRAS, HRAS or NRAS protein by contacting the protein with an effective amount of a compound of the present disclosure. Modulation can inhibit or activate the activity of the protein. In some embodiments, the present disclosure provides a method of inhibiting the activity of a protein by contacting a G12C and / or G12D mutant KRAS, HRAS or NRAS protein with an effective amount of a compound of the present disclosure in solution. In some embodiments, the present disclosure provides a method of inhibiting the activity of a G12C or G12D mutant KRAS, HRAS or NRAS protein by contacting a cell, tissue or organ expressing the protein of interest. In some embodiments, the present disclosure provides a method of inhibiting the activity of a protein in a subject by administering an effective amount of a compound of the present disclosure to a subject, including but not limited to rodents and mammals (e.g., humans). In some embodiments, the percentage of modulation is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90%. In some embodiments, the percentage of inhibition is greater than 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.

[0204] In some embodiments, the present disclosure provides a method of inhibiting the activity of KRAS, HRAS or NRAS G12C or G12D in a cell by contacting the cell with a compound of the present disclosure in an amount sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the cell. In some embodiments, the present disclosure provides a method of inhibiting the activity of KRAS, HRAS or NRAS G12C in a tissue by contacting the tissue with a compound of the present disclosure in an amount sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the tissue. In some embodiments, the present disclosure provides a method of inhibiting the activity of KRAS, HRAS or NRAS G12C in an organ by contacting the organ with a compound of the present disclosure in an amount sufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in the organ. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS or NRAS G12C activity in an animal by contacting the animal with a compound of the present disclosure in an amount sufficient to inhibit KRAS, HRAS or NRAS G12C activity in the animal. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS or NRAS G12C activity in a mammal by contacting the mammal with a compound of the present disclosure in an amount sufficient to inhibit KRAS, HRAS or NRAS G12C activity in the mammal. In some embodiments, the present disclosure provides a method of inhibiting KRAS, HRAS or NRAS G12C activity in a human by contacting the human with a compound of the present disclosure in an amount sufficient to inhibit KRAS, HRAS or NRAS G12C or G12D activity in the human. The present disclosure provides methods of treating a disease mediated by activity of KRAS, HRAS, or NRAS G12C in a subject in need of such treatment.The present disclosure provides methods of treating a disease mediated by activity of KRAS, HRAS, or NRAS G12D in a subject in need of such treatment.

[0205] Combination therapy: The present disclosure also provides a method of combination therapy, in which agents known to regulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes, are used in combination with the compounds of the present disclosure or pharma- ceutically acceptable salts thereof. In one embodiment, such treatment includes, but is not limited to, the combination of one or more compounds of the present disclosure with chemotherapeutic agents, therapeutic antibodies, and radiation therapy to produce synergistic or additive therapeutic effects. In one embodiment, the method further comprises administering a therapeutically effective amount of an additional pharma- ceutical active compound to a patient in need thereof. In another embodiment, the method comprises administering a compound of the present invention prior to administration of at least one additional pharma- ceutical active compound. In another embodiment, the method comprises administering a compound of the present invention simultaneously with administration of at least one additional pharma- ceutical active compound. In another embodiment, the method comprises administering a compound of the present invention after administration of at least one additional pharma- ceutical active compound.

[0206] Currently, many chemotherapeutic agents are known in the art and can be used in combination with the compounds of the present disclosure. In some embodiments, the chemotherapeutic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antihormones, angiogenesis inhibitors, and antiandrogens. Non-limiting examples are chemotherapeutic agents, cytotoxic agents, and non-peptide small molecules such as Gleevec® (imatinib mesylate), Kyprolis® (carfilzomib), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), venetoclax, and adriamycin, as well as a host of chemotherapeutic 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; ethylenimines and methylameramines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobembitine, phenesterine, prednimustine, trofosfamide, and uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine;Aclacinomycin, Actinomycin, Ausramycin, Azaserine, Bleomycin, Cactinomycin, Calicheamicin, Carabicin, Carminomycin, Carzinophilin, Casodex™, Chromomycin, Dactinomycin, Daunorubicin, Detorubicin, 6-Diazo-5-oxo-L-norleucine, Doxorubicin, Epirubicin, Esorubicin, Idarubicin, Marcellomycin, Mitomycin, Mycophenolic acid, Nogalamycin, Olivomycin, Peplomycin, Potofilomycin, Puromycin, Ketomycin, Antibiotics such as ramycin, lodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; metabolic antagonists such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, and trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine; ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxiuridine. Pyrimidine analogues such as sirolimidine, androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; adrenal inhibitors such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as floric acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; amsacrine; bestravcil; bisantrene; edatraxate; defofamine; demecolcine; diazicon; elfomitine; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazo ;Mitoxantrone;Mopidamol;Nitracrine;Pentostatin;Fenameth;Pirarubicin;Podophyllic acid;2-Ethylhydrazide;Procarbazine;PSK;Razoxane;Sizofiran;Spirogermanium;Tenuazonic acid;Triaziquone;2,2',2''-Trichlorotriethylamine;Urethane;Vindesine;Dacarbazine;Mannomustine;Mitobronitol;Mitolactol;Pipobroman;Gacytosine;Arabinoside ("Ara-C");Cyclophosphamide;Thiotepa;Taxanes, such as paclitaxel and docetaxel;Retinoic acid; esperamicin; capecitabine; and pharma- ceutically acceptable salts, acids, or derivatives of any of the above.

[0207] Also included among suitable chemotherapeutic cell regulators are those that act to regulate or inhibit hormone action on tumors, such as tamoxifen (Nolvadex™), raloxifene, aromatase-inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxyphene, ketoxifene, LY. antihormonal agents such as antiestrogens, including 117018, onapristone, and toremifene (Fareston); and antiandrogens such as flutamide, nilutamide, bicalutamide, luprolide, 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); the topoisomerase inhibitor RFS 2000; and difluoromethylornithine (DMFO).

[0208] If desired, the compounds or pharmaceutical compositions of the present disclosure can be used in combination with other agents such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE, abagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alpharazine, alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, anti-CD22 immunotoxins, antineoplastic drugs, antineoplastic herbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW 2992, biricoderm, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), calyculin, cell cycle non-specific antitumor agent, dichloroacetic acid, discodermolide, elsamitrucin, enocitabine, epothilone, eribulin, everolimus, exatecan, exisulind, ferruginol, forodesine, fosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulven, laniquidar, larotaxel, lenalidomide, lucantone, lurtotecan, mafosfamide, mitozolomide , nafoxidine, nedaplatin, olaparib, ortaxel, PAC-1, pawpaw, pixantrone, proteasome inhibitors, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126 or zosuquidar.

[0209] The present disclosure further relates to a method of using the compounds or pharmaceutical compositions provided herein in combination with radiation therapy to inhibit abnormal cell proliferation or treat hyperproliferative disorders in a mammal. Methods for administering radiation therapy are known in the art, and these methods can be used in the combination therapy described herein. The administration of the compounds of the present disclosure in this combination therapy can be determined as described herein.

[0210] Radiation therapy can be administered by one of several methods, or a combination of these methods, including, but not limited to, external beam therapy, internal radiation therapy, implant irradiation, stereotactic radiosurgery, total body radiation therapy, radiotherapy, and permanent or temporary interstitial brachytherapy. As used herein, the term "brachytherapy" refers to radiation therapy delivered by a spatially restricted radioactive material inserted into the body at or near the site of a tumor or other proliferative tissue disease. This term is intended to include, but is not limited to, exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioisotopes of Lu). Suitable radiation sources for use as cell regulators of the present disclosure include both solid and liquid. As non-limiting examples, the radioactive source can be a radionuclide such as I-125, I-131, Yb-169, Ir-192, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma rays, or other therapeutic rays. The radioactive material can be a fluid made from any solution of the radionuclide, such as a solution of I-125 or I-131, or the radioactive fluid can be produced using a slurry of a suitable fluid containing microparticles of a solid radionuclide such as Au-198, Y-90, etc. Additionally, the radionuclide can be embodied as a gel or radioactive microspheres.

[0211] The compounds or pharmaceutical compositions of the present disclosure can be used in combination with an amount of one or more substances selected from anti-angiogenic agents, signal transduction inhibitors, anti-proliferative agents, glycolytic inhibitors, or autophagy inhibitors.

[0212] 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 with the compounds of the present disclosure and the pharmaceutical compositions described herein. 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 useful matrix metalloproteinase inhibitors are disclosed in WO 96 / 33172, WO 96 / 27583, EP 0818442, EP 1004578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, EP 98 / 34915, WO 98 / 33768, WO 98 / 30566, EP 606046, EP 931788, US Pat. No. 5,993,229, and US Pat. No. 5,993,229. Nos. 90 / 05719, 99 / 52910, 99 / 52889, 99 / 29667, 99 / 007675, EP 1786785, 1181017, U.S. Patent Application Publication No. 20090012085, U.S. Patent Nos. 5,863,949, 5,861,510, and EP 0780386, all of which are incorporated herein by reference in their entireties. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and / or AMP-9 relative to other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).Some specific examples of MMP inhibitors useful in the present disclosure are AG-3340, RO32-3555, and RS13-0830.

[0213] The compound also includes acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim, ARGLABIN, arsenic trioxide, BAM 002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030 (Dong-A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon α, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emiteflu, epirubicin, epoetin β, etoposide phosphate, exemestane, exisulind, fado lozole, filgrastim, finasteride, fudrabin phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumabzogamicin, dimeracil / oteracil / tegafur combination, glycopin, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronate, idarubicin (imiquimod, interferon alpha, interferon alpha, natural interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-N1, interferon alpha- n3, interferon alfacon-1, interferon α, natural interferon β, interferon β-1a, interferon β-1b, interferon gamma, natural interferon gamma-1a, natural interferon gamma-1b, interleukin-1β, iobenguane, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leucocyte alpha interferon, leuprorelin, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprocol, melasoprol, metoclopramide, mifepristone, miltefosine, millimostim, mismatched double-stranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis-stimulating protein, NSC 631570Octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronate, pegaspargase, peginterferon alpha-2b, pentosan polysulfate sodium, pentostatin, picibanil, pirarubicin, rabbit antithymocytoma polyclonal antibody, polyethylene glycol interferon alpha-2a, porfimer sodium, raloxifene, raltitrexed, rasbriembodiment, rheniumRe 186Etidronate, RIIretinamide, rituximab, romurtide, samarium (153 Sm) lexidronam, sargramotim, sizofiran, sobuzoxane, sonermin, strontium-89 chloride, suramin, tasonermin, 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, Virulizin, zinostatin stimalamer or zoledronic acid; abarelix;AE 941 (Aeterna), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, dexaminoglutethimide, diazicon, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2, iproxifen, LDI 200 (Milkhaus), religistim, lintuzumab, CA 125 MAb (Biomira), Cancer MAb (Nihon Yakuhin Kaihatsu Co., Ltd.), HER-2 and Fc MAb (Medarex), Idiotypic 105AD7 MAb (CRC Technology), Idiotypic CEA MAb (Trilex), LYM-1-Iodine-131 MAb (Techniclone), Polymorphic Epithelial Mucin-Yttrium-90 MAb (Antisoma), Marimastat, Menogaril, Mitsumomab, Motexafine Gadolinium, MX 6 (Galderma), Nelarabine, Nolatrexed, P 30 Protein, Pegvisomant, Pemetrexed, Porfiromycin, Prinomastat, RL 0903 (Shire), Rubitecan, Satraplatin, Sodium Phenylacetate, Sparfosic Acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA It may also be used in combination therapy with other anticancer drugs such as 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyl etiopurpurin, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), melanoma oncolysate vaccine (New York Medical College), viral melanoma lysate vaccine (Royal Newcastle Hospital) or valspodar;

[0214] The compounds of the present invention may further be used in conjunction with VEGFR inhibitors. The following patents and patent applications: U.S. Pat. No. 6,258,812, U.S. Patent Application Publication No. 2003 / 0105091, WO 01 / 37820, U.S. Pat. No. 6,235,764, WO 01 / 32651, U.S. Pat. No. 6,630,500, U.S. Pat. No. 6,515,004, U.S. Pat. No. 6,713,485, U.S. Pat. No. 5,521,184, U.S. Pat. No. 5,770,599, U.S. Pat. No. 5,747,498, WO 02 / 68406, WO 02 / 66470 and other compounds described in U.S. Pat. Nos. 02 / 55501, 04 / 05279, 04 / 07481, 04 / 07458, 04 / 09784, 02 / 59110, 99 / 45009, 00 / 59509, 99 / 61422, U.S. Pat. No. 5,990,141, WO 00 / 12089, and WO 00 / 02871 can be used in combination therapy.

[0215] In some embodiments, the combination comprises a composition of the present invention in combination with at least one antiangiogenic agent.Agents include, but are not limited to, in vitro synthetically prepared chemical compositions, antibodies, antigen-binding regions, radionuclides, and combinations and conjugates thereof.Agents may be agonists, antagonists, allosteric regulators, toxins, or more generally act to inhibit or stimulate their targets (e.g., activate or inhibit receptors or enzymes), thereby promoting cell death or halting cell proliferation.

[0216] Exemplary anti-angiogenesis agents include ERBITUX™ (IMC-C225), KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., antibodies or antigen binding regions that specifically bind to VEGF or a soluble VEGF receptor or its ligand binding region) such as AVASTIN™ or VEGF-TRAP™, and anti-VEGF receptor drugs (e.g., antibodies or antigen binding regions that specifically bind to), EGFR inhibitors (e.g., antibodies or antigen binding regions that specifically bind to), such as Vectibix (panitumumab), IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-Ang1 and anti-Ang2 agents (e.g., antibodies or antigen binding regions that specifically bind to them or to their receptors, e.g., Tie2 / Tek), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind to them). The pharmaceutical compositions of the invention can also include one or more agents (e.g., antibodies, antigen-binding regions, or soluble receptors) that specifically bind and inhibit the activity of a growth factor, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor) that specifically bind to the receptor "c-met" and antibodies or antigen-binding regions thereof.

[0217] Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (Ceretti et al., U.S. Patent Application Publication No. 2003 / 0162712; U.S. Patent No. 6,413,932), anti-TWEAK agents (e.g., antibodies or antigen binding regions that specifically bind or soluble TWEAK receptor antagonists; Wiley, U.S. Patent No. 6,727,225), ADAM disintegrin domains that antagonize the binding of integrins to their ligands (Fanslow et al., U.S. Patent Application Publication No. 2002 / 0042368), anti-ep h-receptor and / or anti-ephrin antibodies or antigen binding regions (see U.S. Pat. Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124 and members of their patent families) and anti-PDGF-BB antagonists (e.g., antibodies or antigen binding regions that specifically bind), as well as antibodies or antigen binding regions that specifically bind to PDGF-BB ligands and PDGFR kinase inhibitors (e.g., antibodies or antigen binding regions that specifically bind thereto).

[0218] Further anti-angiogenic / anti-tumor agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 770622); pegaptanib octasodium (Gilead Sciences, USA); Alphastatin (BioActa, UK); M-PGA (Celgene, USA, U.S. Pat. No. 5,712,291); ilomastat (Arriva, USA, U.S. Pat. No. 5,892,112); emaxanib (Pfizer, USA, U.S. Pat. No. 5,792,783); vatalanib (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); anecortave acetate (Alcon, USA); α-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands); DAC: antiangiogenic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko Bio, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP 970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen E-fragment (BioActa, UK); Angiogenesis inhibitors (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA);SC-236 (Pfizer, USA);ABT-567 (Abbott, USA);metastatin (EntreMed, USA);angiogenesis inhibitors (Tripep, Sweden);maspin (Sosei Group, Japan);2-methoxyestradiol (Oncology Sciences Corporation, USA);ER-68203-00 (IVAX, USA);Benefin (Lane Labs, USA);Tz-93 (Tsumura, Japan);TAN-1120 (Takeda Pharmaceutical, Japan);FR-111142 (Fujisawa Pharmaceutical Co., Japan, JP 02233610); Platelet factor 4 (RepliGen, USA, EP 407122); Vascular endothelial growth factor antagonists (Borean, Denmark); Bevacizumab (pINN) (Genentech, USA); Angiogenesis inhibitors (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); Second generation MAb, α5β3 integrin (Applied Molecular Evolution, USA and MedImmune, USA); Gene therapy for retinopathy (Oxford BioMedica, UK); Enzastaurin hydrochloride (USAN) (Lilly, USA); CEP7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy);angiogenesis inhibitors (Alchemia, Australia);VEGF antagonists (Regeneron, USA);rBPI 21 and BPI-derived antiangiogenic agents (XOMA, USA);PI 88 (Progen, Australia);cilengitide (pINN) (Merck KGaA, German;Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA);cetuximab (INN) (Aventis, France);AVE 8062 (Ajinomoto, Japan);AS 1404 (Cancer Research Laboratory, New Zealand);SG 292 (Telios, USA);endostatin (Boston Children's Hospital, USA);ATN 161 (Attenuon, USA);angiostatin (Boston Children's Hospital, USA);2-methoxyestradiol (Boston Children's Hospital, USA);ZD 6474 (AstraZeneca, UK); ZD 6126 (Angiogene Pharmaceuticals, UK); PPI 2458 (Praecis, USA);AZD 9935 (AstraZeneca, UK); AZD 2171 (AstraZeneca, UK); vatalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); tissue factor pathway inhibitor (EntreMed, USA); pegaptanib (Pinn) (Gilead Sciences, USA); xanthrizol (Yonsei University, South Korea); gene-based 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, USA); troponin I (Harvard University, USA); SU 6668 (SUGEN, USA); OXI 4503 (OXiGENE, USA); o-guanidine (Dimensional Pharmaceuticals, USA);motuporamine C (British Columbia University, Canada);CDP 791 (Celltech Group, UK);atiprimod (pINN), (GlaxoSmithKline, UK);E 7820 (Eisai, Japan);CYC 381 (Harvard University, USA);AE 941 (Aeterna, Canada);vaccines, angiogenesis (EntreMed, USA);urokinase plasminogen activator inhibitors (Dendreon, USA);oglufanide (pINN) (Melmotte, USA);HIF-1α inhibitors (Xenova, UK);CEP 5214 (Cephalon, USA);BAY RES 2622 (Bayer, Germany);angiocidin (InKine, USA);A6 (Angstrom, USA);KR 31372 (Korea Research Institute of Chemical Technology, South Korea);GW 2286(GlaxoSmithKline, UK);EHT 0101(ExonHit, France);CP 868596 (Pfizer, USA); CP 564959 (OSI, USA); CP 547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN 633 (Kirin Brewery, Japan); drug delivery system, intraocular, 2-methoxyestradiol (EntreMed, USA); Anginex (Maastricht University, Netherlands and 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);MAb, KDR (ImClone Systems, USA);MAb, α5β1 (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);combretastatin A4 prodrug (Arizona State University, USA);chondroitinase AC (IBEX, Canada);BAY RES 2690 (Bayer, Germany);AGM 1470 (Harvard University, USA, Takeda Pharmaceutical, Japan and TAP, USA);AG 13925 (Agouron, USA);tetrathiomolybdate (University of Michigan, USA);GCS 100 (Wayne State University, USA)CV 247 (Ivy Medical, UK);CKD 732 (Chong Kun Dang, South Korea); MAb, vascular endothelial growth factor (Xenova, UK);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 Pharmaceutical, Japan); VEGF receptor modulator (Pharmacopeia, USA); VE-cadherin-2 antagonist (ImClone Systems, USA); vasostatin (National Institutes of Health, USA); vaccine, Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); tumstatin (Beth Israel Hospital, USA); cleaved soluble FLT1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA).

[0219] Autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4 imidazole-carboxamide riboside (AICAR), okadaic acid, autophagy-inhibiting algal toxins that inhibit type 2A or type 1 protein phosphatases, analogs of cAMP, and drugs that increase cAMP levels, such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Additionally, antisense or siRNA that inhibit the expression of proteins, including but not limited to ATG5 (involved in autophagy), may also be used.

[0220] Additional pharma- ceutically active compounds / drugs that may be used in the treatment of cancer and that may 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 386; AMG 479; AMG 655; AMG 745; AMG 951; and AMG 706 or pharma- ceutically acceptable salts thereof.

[0221] In certain embodiments, the compositions provided herein are administered in combination with a chemotherapeutic agent. Suitable chemotherapeutic agents include natural products such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), paclitaxel, epidipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, doxorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparagine-containing systemic inhibitors), and the like. L-asparaginase, which efficiently metabolizes and removes cells that do not have the ability to synthesize their own asparagine; antiplatelet agents; antiproliferative / antimitotic alkylating agents such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide and analogs, melphalan, and chlorambucil); ethylenimines and methylmelamines (e.g., hexamethylmelamine and thiotepa); CDK inhibitors (e.g., seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, , P27-00, AT-7519, RGB286638, and SCH727965), alkylsulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs, and streptozocin), trazene-dacarbazine (DTIC), antiproliferative / antimitotic antimetabolites such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercury, bromine, benzodiazepine, and benzoic acid). captopurine, thioguanine, pentostatin, and 2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoylanilide hydroxamic acid apicidan, vorinostat, LBH589, romidepsin, ACY-1215 and panobinostat), mTor inhibitors (e.g., temsirolimus, everolimus, ridaforolimus and sirolimus), KSP (Eg5) inhibitors (e.g., Array 520), DNA binders (e.g., Zalipsis), PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), multikinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen), and hormonal agonists such as luteinizing hormone releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide, and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN 163L), Aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38)), anti-CS1 (e.g., elotuzumab), HSP90 inhibitors (e.g., 17 AAG and KOS 953), P13K / Akt inhibitors (e.g., Perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., Enzastaurin), FTIs (e.g., Zarnestra™), anti-CD138 (e.g., BT062), Torc1 / 2 specific kinase inhibitors (e.g., INK128), kinase inhibitors (e.g., GS-1101), ER / UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1 / 2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), BCL-2 antagonists. Other chemotherapeutic agents may include mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, or analogs or derived variants of any of the above.

[0222] The compounds of the present invention may also be used in combination with radiation therapy, hormone therapy, surgery, and immunotherapy, all of which are well known to those skilled in the art.

[0223] In certain embodiments, the pharmaceutical compositions provided herein are administered in combination with a steroid. Suitable steroids include 21-acetoxypregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximethasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandre The therapeutically active agents may include, but are not limited to, cyclosporine ... Examples of drugs that can be used to treat nausea include the following: dronabinol; granisetron; metoclopramide; ondansetron; and prochlorperazine; or pharma- ceutically acceptable salts thereof.

[0224] The compounds of the present invention can also be used in combination with additional pharmacologic active compounds that interrupt or inhibit RAS-RAF-ERK or PI3K-AKT-TOR signaling pathways.In other such combinations, the additional pharmacologic active compounds are PD-1 and PD-L1 antagonists.The compounds or pharmaceutical compositions of the present disclosure can 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 such as (RMC-4630), proteasome inhibitors, and immunotherapies including monoclonal antibodies, immunomodulatory imids (IMiDs), anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40 agents, GITR agonists, CAR-T cells, and BiTEs.

[0225] EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux), panitumumab (Vectibix), zalutumumab, nimotuzumab, and matuzumab. Small molecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), and more recently lapatinib (TykerB). See, e.g., Yan L, et.al., Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic Antibody Development, BioTechniques 2005;39(4):565-8 and Paez JG, et.al., EGFR Mutations In Lung Cancer Correlation With Clinical Response To Gefitinib Therapy, Science 2004;304(5676):1497-500.

[0226] Non-limiting examples of small molecule EGFR inhibitors include the following: European Patent Application Publication No. 520722, published December 30, 1992; European Patent Application Publication No. 566226, published October 20, 1993; International Patent Application Publication No. WO 96 / 33980, published October 31, 1996; U.S. Patent No. 5,747,498, issued May 5, 1998; International Patent Application Publication No. WO 96 / 30347, published October 3, 1996; European Patent Application Publication No. 787772, published August 6, 1997; International Patent Application Publication No. WO 96 / 30347, published August 21, 1997; Publication No. 97 / 30034; Publication No. 97 / 30044, published August 21, 1997; Publication No. 97 / 38994, published October 23, 1997; Publication No. 97 / 49688, published December 31, 1997; Publication No. 837063, published April 22, 1998; Publication No. WO 98 / 02434, published January 22, 1998; Publication No. WO 97 / 38983, published October 23, 1997; Publication No. WO 95 / 19774, published July 27, 1995; Publication No. WO 97 / 2000 / 02 / 1996 ... German Patent Application Publication No. 95 / 19970, published April 27, 1997; German Patent Application Publication No. 97 / 13771, published April 17, 1997; German Patent Application Publication No. 98 / 02437, published January 22, 1998; German Patent Application Publication No. 98 / 02438, published January 22, 1998; German Patent Application Publication No. 97 / 32881, published September 12, 1997; German Patent Application Publication No. 19629652, published January 29, 1998; International Patent Application Publication No. WO 98 / 33798, published August 6, 1998; International Patent Application Publication No. WO 97 / 32880, published September 12, 1997; European Patent Application Publication No. 97 / 32880, published September 12, 1997; European Patent Application Publication No. 682027, published November 15, 1995; International Patent Application Publication No. WO 97 / 02266, published January 23, 1997; International Patent Application Publication No. WO 97 / 27199, published July 31, 1997; International Patent Application Publication No. WO 98 / 07726, published February 26, 1998; International Patent Application Publication No. WO 97 / 34895, published September 25, 1997; International Patent Application Publication No. WO 96 / 31510, published October 10, 1996; International Patent Application Publication No. WO 98 / 14449, published April 9, 1998;No. 98 / 14450 published April 9, 1998; No. 98 / 14451 published April 9, 1998; No. 95 / 09847 published April 13, 1995; No. 97 / 19065 published May 29, 1997; No. 98 / 17662 published April 30, 1998; U.S. Patent No. 5,789,427 issued August 4, 1998; U.S. Patent No. 5,650,415 issued July 22, 1997; No. 5,656,643, published Aug. 12; WO 99 / 35146, published July 15, 1999; WO 99 / 35132, published July 15, 1999; WO 99 / 07701, published February 18, 1999; and WO 92 / 20642, published November 26, 1992, and all pharma- ceutically acceptable salts and solvates of said EGFR inhibitors. Additional non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler, P., 1998, Exp. Opin. Ther. Patents 8(12):1599-1625.

[0227] Antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation 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, SM, 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 the monoclonal antibody Mab E7.6.3 (Yang, 1999, supra), or Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment having the binding specificity thereof.

[0228] MEK inhibitors include, but are not limited to, trametinib (Mekinist®), CI1040, AZD6244, PD318088, PD98059, PD334581, RDEA119, ARRY-142886, ARRY438162, and PD325901.

[0229] PI3K inhibitors include wortmannin, 17-hydroxywortmannin analogs described in WO 06 / 044453, 4-[2-(1H-indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine (also known as GDC 0941 and described in WO 09 / 036,082 and WO 09 / 055,730), 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile (BEZ 235 or NVP-BEZ 235, also known as 5-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one ... N'-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfono-hydrazide hydrochloride available from 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-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidine bismesylate), AS-252424 (5-[1-[5-(4-fluoro-2-hydroxy-phenyl)-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 demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477, CUDC-907 and AEZS-136.

[0230] AKT inhibitors include Akt-1-1 (inhibits Akt1) (Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2 (inhibits Ak1 and 2) (Barnett et al. (2005) Biochem. J. 385 (Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05011700); indole-3-carbinol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134 (12 Suppl), 3493S-3498S); perifosine (e.g., which interferes with membrane localization of Akt; Dasmahapatra et al. (2004) Clin. Cancer Res. 10(15), 5242-52, 2004); phosphatidylinositol ether lipid analogues (e.g., 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).

[0231] TOR inhibitors include, but are not limited to, inhibitors including 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, such as: CCI-779 (temsirolimus), RAD001 (everolimus; WO 9409010) and AP23573; rapalogs, such as those disclosed in WO 98 / 02441 and WO 01 / 14387, such as AP23573, AP23464 or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3 [-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also known as CC1779), 40-epi-(tetrazolyl)-rapamycin (also known as ABT578), 32-deoxorapamycin, 16-pentynyloxy-32(S)-dihydrorapanycin and other derivatives disclosed in WO 05005434; U.S. Pat. No. 5,258,389; WO 05005436; Patent Publication Nos. 94 / 090101, 92 / 05179, U.S. Pat. Nos. 5,118,677, 5,118,678, 5,100,883, 5,151,413, and 5,120,842, International Publication Nos. 93 / 111130, 94 / 02136, 94 / 02485, 95 / 14023, and 94 / rapamycin and its derivatives, including phosphorus-containing rapamycin derivatives (e.g., WO 02136, WO 95 / 16691, WO 96 / 41807, WO 96 / 41807 and derivatives disclosed in U.S. Pat. No. 5,256,790; rapamycin and its derivatives, including 4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional Patent Application No. 60 / 528,340).

[0232] MCl-1 inhibitors include, but are not limited to, AMG-176, MIK665 and S63845.Myeloid cell leukemia-1 (MCL-1) protein is one of the important anti-apoptotic members of B-cell lymphoma-2 (BCL-2) protein family.Overexpression of MCL-1 is closely related to tumor progression and is closely related to resistance not only to conventional chemotherapy but also to targeted therapy, including BCL-2 inhibitors such as ABT-263.

[0233] SHP inhibitors include, but are not limited to, SHP099.

[0234] Proteasome inhibitors include, but are not limited to, Kyprolis® (carfilzomib), Velcade® (bortezomib), and oprozomib.

[0235] Immunotherapies include, but are not limited to, anti-PD-1 agents, anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1 agents, and anti-OX40 agents.

[0236] Monoclonal antibodies include, but are not limited to, Darzalex® (daratumumab), Herceptin® (trastuzumab), Avastin® (bevacizumab), Rituxan® (rituximab), Lucentis® (ranibizumab), and Eylea® (aflibercept).

[0237] Immunomodulatory imide drugs (IMiDs) are a class of immunomodulators (drugs that modulate the immune response) that contain an imide group. The IMiD class includes thalidomide and its analogs (lenalidomide, pomalidomide, and apremilast).

[0238] Exemplary anti-PD-1 antibodies and methods for their use 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 (WO 2006 / 121168 A1), each of which is expressly incorporated herein by reference, including pembrolizumab (Keytruda®), nivolumab (Opdivo®), Yervoy™ (ipilimumab) or tremelimumab (against CTLA-4), galiximab (against B7.1), BMS-936558 (against PD-1), MK-3475 (against PD-1), AMP224 (against B7DC), BMS-936559 (against B7-H1), MPDL3280A (against B7-H1), MEDI-570 (against ICOS), AMG 404, AMG557 (against B7H2), MGA271 (against B7H3), IMP321 (against LAG-3), BMS-663513 (against CD137), PF-05082566 (against CD137), CDX-1127 (against CD27), anti-OX40 (Providence Health Services), huMAbOX40L (against OX40L), atacicept (against TACI), CP-870893 (against CD40), lucatumumab (against CD40), decatuzumab (against CD40), muromonab-CD3 (against CD3), ipilumumab (against CTLA-4). Immunotherapies also include engineered T cells (e.g., CAR-T cells) and bispecific antibodies (e.g., BiTEs).

[0239] GITR agonists include GITR fusion proteins described in U.S. Pat. No. 6,111,090box.c, European Patent No. 090505B1, U.S. Pat. No. 8,586,023, WO 2010 / 003118 and WO 2011 / 090754, or GITR fusion proteins described in, for example, U.S. Pat. No. 7,025,962, European Patent No. 1947183B1, U.S. Pat. No. 7,812,135, U.S. Pat. No. 8,388,967, U.S. Pat. No. 8,591,886, European Patent No. 1866339, WO 2011 / 028683, WO 2013 / 039 Nos. 954, 2005 / 007190, 2007 / 133822, 2005 / 055808, 99 / 40196, 2001 / 03720, 99 / 20758, 2006 / 083289, 2005 / 115451, U.S. Pat. No. 7,618,632, and WO 2011 / 051726, and other such GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies).

[0240] The compounds described herein may be used in combination with the agents disclosed herein or other suitable agents depending on the condition being treated. Thus, in some embodiments, one or more compounds of the present disclosure will be co-administered with the other agents described above. When used in combination therapy, the compounds described herein are administered simultaneously with the second agent or separately. This combination administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration of separate dosage forms, and separate administration. That is, the compounds described herein and any of the agents described above may be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds disclosed herein and any of the agents described above may be administered simultaneously, where both agents are in separate formulations. In other alternative embodiments, the compounds disclosed herein may be administered followed by any of the agents described above, or in the reverse order. In some embodiments of the separate administration protocol, the compounds disclosed herein and any of the agents described above are administered within minutes, or within hours, or within days.

[0241] As one aspect of the present invention contemplates treating a disease / condition with a combination of pharma- ceutically 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 comprises two separate pharmaceutical compositions: a compound of the present invention and a second pharmaceutical compound. The kit comprises containers, such as separate bottles or separate foil packets, for containing the separate compositions. Other examples of containers include syringes, boxes, and bags. In some embodiments, the kit comprises instructions for using the separate components. The kit form is particularly advantageous when these separate components are preferably administered in different dosage forms (e.g., oral and parenteral), administered at different dosage intervals, or when the prescribing physician desires to titrate the individual components of the combination.

[0242] Synthesis of the Compounds Disclosed herein The compounds disclosed herein can be synthesized through a number of specific methods. The examples outlining specific synthetic routes and the following general schemes are intended to provide guidance to a synthetic chemist of ordinary skill, who will readily understand that solvents, concentrations, reagents, protecting groups, order of synthetic steps, times and temperatures, etc., can be varied as necessary, well within the skill and judgment of the skilled artisan. WO 2018 / 119183, WO 2018 / 217651 (International Application PCT / US / 2018 / 033714 and corresponding application also referred to as US Patent Publication No. 2019 / 051291) and WO 2019 / 051291 can be used in the present invention and are incorporated herein in their entirety for all purposes.

[0243] The following method described in WO 2018 / 217651 and its corresponding US 2018 / 0334454 is explicitly recited herein to prepare the intermediate (S)-tert-butyl-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. The solution of (S)-tert-butyl 4-(6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate is then used to prepare a novel embodiment of the present invention. [ka]

[0244] Step 1: 2-Isopropyl-4-methylpyrimidin-3-amine (Intermediate R). To a slurry of 3-amino-2-bromo-4-picoline (360 mg, 1.9 mmol, Combi-Blocks, San Diego, CA, USA) in THF (4 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (79 mg, 0.10 mmol) complexed with dichloromethane. The resulting slurry was deoxygenated with argon for 2 min and then 2-propylzinc bromide (0.5 M solution in THF, 5.40 mL, 2.7 mmol, Sigma-Aldrich, St. Louis, MO) was added. The resulting solution was heated at 60° C. for 17 h, then the heat was removed and the reaction was allowed to cool to room temperature. The reaction mixture was quenched with water (10 mL) and 1N NaOH solution (20 mL) and then extracted with EtOAc (2×). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-15% MeOH / DCM) to give 2-isopropyl-4-methylpyridin-3-amine. 1 H NMR (400MHz, DMSO-d 6 )δ ppm 7.66(d,J=4.6Hz,1H),6.78(d,J=4.8Hz,1H),4.72(br s,2H),3.14-3.25(m,1H),2.08(s,3H),1.14(d,J=6.8Hz,6H).m / z(ESI,+ve ion):151.1(M+H) + .

[0245] Step 2: 2,5,6-trichloro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. To a -78°C slurry of 2,5,6-trichloronicotinamide (Intermediate P, 3.10 g, 13.8 mmol) in THF (46 mL) was added oxalyl chloride (2 M solution in DCM, 7.4 mL, 14.7 mmol) slowly via syringe. The resulting slurry was heated at 60°C for 3.5 h, then the heat was removed and the reaction cooled to -78°C. Triethylamine (6.0 mL, 42.6 mmol) was added via cannula followed by a solution of 2-isopropyl-4-methylpyridin-3-amine (Intermediate R, 2.12 g, 14.1 mmol). The resulting slurry was allowed to warm to room temperature and stirred for 1 h, then partitioned between water (120 mL) and EtOAc (175 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in 9:1 heptane / EtOAc and filtered. The filtered solid was collected to give 2,5,6-trichloro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide. 1 H NMR (400MHz, DMSO-d 6 )δ ppm 11.31(s,1H),9.54(s,1H),8.66(s,1H),8.34(d,J=4.8Hz,1H),7.16(d,J=5. 0Hz,1H),3.24-3.33(m,1H),2.22(s,3H),1.17(d,J=6.6Hz,6H).m / z(ESI,+ve ion):400.9(M+H) + .

[0246] Step 3: 6,7-Dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione. To an ice-cold solution of 2,5,6-trichloro-N-((2-isopropyl-4-methylpyridin-3-yl)carbamoyl)nicotinamide (4.71 g, 11.7 mmol) in THF (55 mL) was added KHMDS (1 M solution in THF, 23.5 mL, 23.5 mmol) slowly via syringe. After 10 min, the ice bath was removed and the resulting solution was stirred at room temperature for an additional 30 min. The reaction was quenched with saturated aqueous ammonium chloride solution (125 mL) and extracted with EtOAc (250 mL). The organic layer was washed with brine (1×), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-11% MeOH / DCM) to give 6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione. 1 H NMR (400MHz, DMSO-d 6 )δ ppm 12.27(br s,1H),8.59(s,1H),8.52(d,J=5.0Hz,1H),7.28(d,J=5.0Hz,1H),2.82-2.92(m,1H),2.04(s,3H),1.08(d,J=6.6Hz,3H),1.01(d,J= 6.8Hz,3H).m / z(ESI,+ve ion):365.0(M+H) + .

[0247] Step 4: 4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. To a slurry of 6,7-dichloro-1-(2-isopropyl-3-methylpyridin-yl)pyrido[2,3-d]pyrimidin-2,4(1H,3H)-dione (2.52 g, 6.9 mmol) in acetonitrile (45 mL) was added DIPEA (1.80 mL, 10.3 mmol), followed by phosphorus oxychloride (1.58 mL, 10.3 mmol) slowly added via syringe. The resulting mixture was heated at 80° C. for 1.75 h, then cooled to room temperature and concentrated to give 4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one, which was used in the next step without further purification.

[0248] Step 5: (S)-tert-Butyl-4-(6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. To a solution of 4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (2.64 g, 6.9 mmol) in THF (40 mL) was added DIPEA (3.61 mL, 20.7 mmol) followed by (S)-4-N-Boc-2-methylpiperazine (2.07 g, 10.3 mmol, Combi-Blocks, Inc., San Diego, CA, USA). The resulting solution was stirred at room temperature for 1.5 h, after which ice water (60 mL) was added. The mixture was stirred for an additional 5 min and then extracted with EtOAc (3×). The combined organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-11% MeOH / DCM) to give (S)-tert-butyl 4-(6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. 1H NMR(400MHz,DMSO-d 6 )δ ppm 8.46(dd,J=11.6,5.2Hz,2H),7.25(d,J=4.8Hz,1H),4.79-4.93(m,1H),4.10-4.24(m,1H),3.87-4.05(m,1H),3.77-3.87(m,1H),3.62-3.76(m,1H),2.99-3.25(m,2H),2.55-2.69(m,1H),1.94(d,J=2.5Hz,3H),1.45(s,9H),1.32(br t,J=5.7Hz,3H),1.06(d,J=6.8Hz,3H),1.00(d,J=6.6Hz,3H).m / z(ESI,+ve イオン):547.2(M+H) + .

[0249] Step 6: (S)-tert-Butyl-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. To a solution of (S)-tert-butyl 4-(6,7-dichloro-1-(2-isopropyl-4-methylpiperazin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (1.02 g, 1.8 mmol) in 1,4-dioxane (17 mL) was added potassium acetate (914 mg, 9.3 mmol) and (2-fluorophenyl)boronic acid (313 mg, 2.2 mmol, Sigma-Aldrich, St. Louis, MO, USA). The mixture was sparged with argon and then [1,1'-bis(diphenylphosphiro)ferrocene]dichloropalladium(II) complexed with dichloromethane (76 mg, 0.093 mmol) was added. The mixture was sparged again with argon and heated at 90°C. After 30 seconds, 3 drops of water were added to the reaction mixture. Heating was continued at 90° C. for 40 minutes, then the reaction was allowed to cool to room temperature. Water (50 mL) and brine (4 mL) were added and the resulting mixture was extracted with EtOAc (2×). The combined organic layers were washed with brine (1×), dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography (eluent: 0-9% MeOH / DCM) to give (S)-tert-butyl 4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate. 1 H NMR (400MHz, DMSO-d 6)δ ppm 8.43(d,J=2.5Hz,1H),8.39(d,J=4.8Hz,1H),7.47-7.55(m,1H),7.16-7.33(m,4H),4.86-4.97(m,1H),4.21-4.30(m,1H),3.9 0-4.06(m,2H),3.80-3.89(m,1H),3.67-3.78(m,1H),3.04-3.16(m,1H),2.65-2.75(m,1H),1.93(s,3H),1.48(s,9H),1.36(br d,J=6.6Hz,3H),1.06(d,J=6.6Hz,3H),0.94(dd,J=6.6,2.1Hz,3H).m / z(ESI,+ve ion):607.0(M+H) + .

[0250] Example 1 (S)-6-Chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyrimidin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine 1-carboxylate (US 20180334454 A1) (500 mg, 0.84 mmol) in dichloromethane (3 mL) was treated with trifluoroacetic acid (1.5 mL, 20.1 mmol) at room temperature and stirred for 15 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one, m / z (ESI, +ve) 507.0 (M+H). + .

[0251] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and chloroacetic acid (156 mg, 1.65 mmol) in DMF (3 mL) at 0° C., DIPEA (0.7 mL, 4.12 mmol) was added followed by TBTU (529 mg, 1.65 mmol). The reaction mixture was stirred at room temperature for 1 h. To the resulting mixture was added water (15 mL) and extracted with EtOAc (15 mL×2). The organic extract was washed with saturated NH 4 Cl (30 mL) and brine (30 mL), and then washed with MgSO 4 The mixture was dried over low-temperature hexanes. The solution was filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (eluent: 0-60% EtOAc-EtOH (3:1) / heptane) to give (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (439 mg, 0.75 mmol, 91% yield, 95% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 ) δ 8.46(brd,J=10.8Hz,1H),8.40(d,J=4.8Hz,1H),7.44-7.59(m,1H),7.25-7.36 (m,2H),7.17-7.24(m,2H),4.87-5.00(m,1H),4.38-4.61(m,2H),4.16-4.36(m, 2H),3.63-4.01(m,2H),3.39-3.53(m,1H),3.10-3.27(m,1H),2.64-2.78(m,1H) ),1.95(s,3H),1.30-1.47(m,3H),1.08(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ ppm -114.03(d,J=6.1Hz,1 F).m / z(ESI,+ve)582.2(M+H) + .

[0252] Example 2 (S)-6-Chloro-4-(4-(2-fluoroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyrimidin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (US 20180334454 A1) (104 mg, 0.17 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 30 minutes. The reaction mixture was concentrated in vacuo to provide (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z(ESI,+ve)507.0(M+H) + .

[0253] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyrimidin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and trifluoroacetic acid (1.0 mL, 13.42 mmol) in DMF (1 mL) was added DIPEA (0.2 mL, 0.86 mmol) followed by TBTU (83 mg, 0.26 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 10 min. To the resulting mixture was added water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was washed with saturated NH 4 Wash with Cl (15 mL) and MgSO 4The mixture was dried over low-temperature hexanes. The solution was filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (eluent: 0-60% EtOAc-EtOH (3:1) / heptane) to give (S)-6-chloro-4-(4-(2-fluoroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (43.4 mg, 0.08 mmol, 44.7% yield, 96% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ8.30-8.52(m,2H),7.46-7.59(m,1H),7.14-7.37(m,4H),5.07-5.44(m,2H),4.91(brs,1H),4.13-4.39(m,2H),3.49-3.86(m, 2H),3.05-3.24(m,1H),2.89(s,2H),1.94(s,3H),1.38(brdd,J=13.8,6.5Hz,3H),1.07(d,J=6.6Hz,3H),0.94(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ ppm -114.02(s,1 F),-228.53(brd,J=248.0Hz,1 F).m / z(ESI,+ve)567.0(M+H) + .

[0254] Example 3 (S)-1-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)propane-1,2-dione [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (96 mg, 0.16 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 10 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0255] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one in DMF (1 mL) was added a mixture of pyruvic acid (13.9 mg, 0.01 mL, 0.16 mmol, Sigma-Aldrich Corporation) and TBTU (102 mg, 0.32 mmol, Advanced ChemTech) in DMF (1 mL) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was purified by HPLC with NaCl. 2 SO 4 The mixture was dried over silica gel. The solution was filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give (S)-1-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)propane-1,2-dione (42.5 mg, 0.07 mmol, 46.6% yield, 97% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d6 )δ 8.44(d,J=4.98Hz,1H),8.39(d,J=4.77Hz,1H),7.46-7.57(m,1H),7.24- 7.35(m,2H),7.14-7.23(m,2H),4.81-5.12(m,1H),4.11-4.42(m,2H),3. 39-3.85(m,3H),2.69(s,2H),2.43(d,J=14.51Hz,3H),1.94(s,3H),1.37 (dd,J=6.84,10.16Hz,3H),1.06(dJ=6.84Hz,3H),0.94(d,J=6.63Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.06(s,1F).m / z(ESI):577.2.0(M+H) + .

[0256] Example 4 (S)-2-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)-2-oxoacetaldehyde [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (207 mg, 0.34 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 10 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0257] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and dl-glyceric acid (181 mg, 0.68 mmol, TCI America) in DMF (1 mL) was added DIPEA (220 mg, 0.3 mL, 1.71 mmol) followed by TBTU (219 mg, 0.68 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 16 h. Additional dl-glyceric acid (181 mg, 0.68 mmol) and TBTU (219 mg, 0.68 mmol) were added and the reaction mixture was stirred for an additional 2 h. The resulting mixture was added with 1 M aqueous HCl (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was discarded. The aqueous solution was basified to pH=9 with 2N NaOH aqueous solution and extracted with EtOAc (20 mL×2). The organic extracts were washed with Na 2 SO 4 The mixture was dried over low-temperature. The solution was filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give 6-chloro-4-((2S)-4-(2,3-dihydroxypropanoyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (78 mg, 0.13 mmol, 38.4% yield, 92% purity) as a yellow solid. m / z (ESI): 595.2.0 (M+H). + .

[0258] A solution of 6-chloro-4-((2S)-4-(2,3-dihydropropanoyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (36 mg, 0.06 mmol) in THF (1.0 mL) / water (0.1 mL) was treated with sodium (meta)periodonate (25.9 mg, 0.12 mmol, Sigma-Aldrich Corporation) at room temperature and stirred for 2.5 h. The resulting mixture was added with water (5 mL) and extracted with DCM (10 mL x 2). The organic extracts were combined and diluted with Na 2 SO 4 The mixture was dried over low-temperature hexanes. The solution was filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (eluent: 0-100% EtOAc / EtOH (3:1) in DCM) to give (S)-2-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)-2-oxoacetaldehyde (26.8 mg, 0.05 mmol, 79% yield, 99% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ 9.46(d,J=12.23Hz,1H),8.42-8.51(m,1H),8.39(d,J=4.77Hz,1H),7.45-7 .57(m,1H),7.13-7.37(m,4H),6.39-6.59(m,1H),4.76-5.31(m,2H),4.27- 4.39(m,1H),4.16-4.25(m,1H),3.56-3.82(m,2H),2.64-2.79(m,1H),1.94 (s,3H),1.26-1.46(m,3H),1.07(d,J=6.63Hz,3H),0.94(d,J=6.63Hz,3H). 19 F NMR(376MHz,DMSO-d6)δ -114.13--113.92(m,1F).m / z(ESI):581.2.0(M+H 2 O) + .

[0259] Example 5 Methyl (S,E)-4-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)-4-oxobut-2-enoate [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (108 mg, 0.18 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 10 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0260] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one in DMF (1 mL) was added a mixture of monomethyl fumarate (34.7 mg, 0.27 mmol, TCI America) and TBTU (114 mg, 0.36 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 15 min. Water (2 mL) was added to the reaction mixture. The resulting precipitate was collected by filtration, washed with water and dried to give methyl (S,E)-4-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazin-1-yl)-4-oxobut-2-enoate (29.3 mg, 0.05 mmol, 26.6% yield, 94% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.44(d,J=17.4Hz,1H),8.39(d,J=5.0Hz,1H),7.47-7.62(m,2H),7.24-7.36(m,2H),7.16-7.23(m ,2H),6.67(dd,J=11.1,15.5Hz,1H),4.96(ddd,J=3.1,6.9,14.9Hz,1H),4.24-4.46(m,2H),3.97-4 .18(m,1H),3.79-3.88(m,1H),3.76(s,3H),3.43-3.60(m,1H),3.11-3.25(m,1H),2.62-2.75(m,1 H),1.94(d,J=2.90Hz,3H),1.35(brd,J=5.18Hz,3H),1.07(d,J=6.6Hz,3H),0.94(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.03(s,1F).m / z(ESI):619.2(M+H) + .

[0261] Example 6 (S)-6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-phenoxyacetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (114 mg, 0.19 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 10 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0262] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimin-2(1H)-one in DMF (1 mL) was added DIPEA (0.16 mL, 0.94 mmol) followed by TBTU (90 mg, 0.28 mmol) at 0° C. The reaction mixture was stirred at 0° C. for 10 min. To the resulting mixture was added water (2 mL). The resulting precipitate was collected by filtration, washed with water, and dried to give a pale yellow solid, which was purified by silica gel chromatography (eluent: 0-100% EtOAc / heptane) to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-phenoxyacetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (92.4 mg, 0.14 mmol, 77% yield, 99% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ ppm 8.44(brd,J=15.3Hz,1H),8.39(d,J=4.8Hz,1H),7.46-7.58(m,1H),7.24- 7.35(m,4H),7.17-7.23(m,2H),6.92-7.04(m,3H),4.79-5.07(m,3H),4.16 -4.38(m,2H),3.40-4.05(m,3H),3.16-3.25(m,1H),2.64-2.78(m,1H),1. 94(s,3H),1.26-1.50(m,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ ppm -114.02(s,1 F).m / z(ESI,+ve)641.1(M+H) + .

[0263] Example 7 (S)-6-Chloro-4-(4-(2-(2,6-dichlorophenoxy)acetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A mixture of (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (Example 1) (88 mg, 0.15 mmol), 99% 2,6-dichlorophenol (49.2 mg, 0.30 mmol) and anhydrous potassium carbonate (41.7 mg, 0.30 mmol) in DMF (1 mL) was stirred and heated at 50° C. for 30 min. The resulting mixture was cooled to 0° C. and water (2 mL) was added. The resulting precipitate was collected by filtration, washed with water, and dried to give a pale yellow solid, which was purified by silica gel chromatography (eluent: 0-100% EtOAc / MeOH in heptane (9:1)) to give (S)-6-chloro-4-(4-(2-(2,6-dichlorophenoxy)acetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (6.2 mg, 8.73 μmol, 5.79% yield, 97% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6)δ 8.46(d,J=11.0Hz,1H),8.39(d,J=5.0Hz,1H),7.53(d,J=8.1Hz,3H),7.17-7 .34(m,5H),4.89-5.02(m,1H),4.84(s,1H),4.70-4.81(m,1H),4.19-4.37(m ,2H),3.42-3.94(m,3H),3.08-3.25(m,1H),2.64-2.75(m,1H),1.94(s,3H), 1.40(brdd,J=13.7,6.6Hz,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ ppm -114.04(s,1 F).m / z(ESI,+ve)709.0 / 711.0(1:1)(M+H) + .

[0264] Example 8 (S)-6-Chloro-4-(4-(2-(2,6-difluorophenoxy)acetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A mixture of (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (Example 1) (50 mg, 0.09 mmol), 2,6-difluorophenol (22.3 mg, 0.17 mmol) and anhydrous potassium carbonate (23.7 mg, 0.17 mmol) in DMF (1 mL) was stirred and heated at 50° C. for 45 min. The resulting mixture was cooled to 0° C. and water (2 mL) was added. The resulting precipitate was collected by filtration, washed with water and dried to give (S)-6-chloro-4-(4-(2-(2,6-difluorophenoxy)acetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (51.4 mg, 0.08 mmol, 89% yield, 95% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.45(brd,J=14.5Hz,1H),8.39(d,J=4.8Hz,1H),7.44-7.58(m,1H),7.24 -7.34(m,2H),7.06-7.23(m,5H),4.97-5.17(m,2H),4.92(brs,1H),4.11- 4.35(m,2H),3.37-3.99(m,3H),3.07-3.24(m,1H),2.64-2.76(m,1H),1.9 4(s,3H),1.32-1.46(m,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.03(s,1 F),-128.21(d,J=17.3Hz,2 F).m / z(ESI,+ve)677.8(M+H) + .

[0265] Example 9 (S)-6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,3,5,6-tetrafluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A mixture of (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (50 mg, 0.09 mmol), 2,3,5,6-tetrafluorophenol (28.5 mg, 0.17 mmol) and anhydrous potassium carbonate (47.4 mg, 0.34 mmol) in DMF (1 mL) was stirred and heated at 50° C. for 45 min. The resulting mixture was cooled to 0° C. and water (2 mL) was added. The resulting precipitate was collected by filtration, washed with water, and dried to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,3,5,6-tetrafluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (39.8 mg, 0.06 mmol, 65.1% yield, 95% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6 )δ ppm 8.45(brd,J=14.9Hz,1H),8.39(d,J=5.0Hz,1H),7.47-7.64(m,2H),7.24- 7.35(m,2H),7.13-7.23(m,2H),5.12-5.41(m,2H),4.87-4.99(m,1H),4.07 -4.38(m,2H),3.35-3.91(m,3H),3.02-3.26(m,1H),2.64-2.77(m,1H),1. 94(s,3H),1.31-1.47(m,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6)δ ppm -114.02(d,J=2.6Hz,1 F),-140.82(dt,J=21.0,7.3Hz,2F),-157.29(td,J=21.2,8.7Hz,2F).m / z(ESI,+ve)713.2(M+H) + .

[0266] Example 10 (S)-6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,3,6-trifluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A mixture of (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (50 mg, 0.09 mmol), 2,3,6-trifluorophenol (38.1 mg, 0.26 mmol) and anhydrous potassium carbonate (47.4 mg, 0.34 mmol) in DMF (1 mL) was stirred and heated at 50° C. for 45 min. The resulting mixture was cooled to 0° C. and water (2 mL) was added. The resulting precipitate was collected by filtration, washed with water and dried to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,3,6-trifluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (47.3 mg, 0.068 mmol, 79% yield, 95% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6)δ ppm 8.45(brd,J=14.7Hz,1H),8.39(d,J=4.8Hz,1H),7.47-7.57(m,1H),7.24-7 .36(m,2H),7.12-7.23(m,4H),5.03-5.34(m,2H),4.92(brd,J=1.2Hz,1H),4 .14-4.36(m,2H),3.38-3.95(m,3H),3.04-3.25(m,1H),2.65-2.76(m,1H),1 .94(s,3H),1.31-1.45(m,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ ppm -114.03(s,1 F),-132.56(ddd,J=26.9,13.0,5.2Hz,1 F),-141.58(dd,J=21.7,13.0Hz,1 F),-151.42(brd,J=21.7Hz,1 F).m / z(ESI,+ve695.2(M+H) + .

[0267] Example 11 (S)-6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,4,6-trifluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A mixture of (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (54 mg, 0.09 mmol), 2,4,6-trifluorophenol (41.1 mg, 0.28 mmol) and anhydrous potassium carbonate (64.0 mg, 0.46 mmol) in DMF (1 mL) was stirred and heated at 50° C. for 60 min. The resulting mixture was cooled to 0° C. and water (2 mL) was added. The resulting precipitate was collected by filtration, washed with water and dried to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,4,6-trifluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (51.3 mg, 0.07 mmol, 80% yield, 94% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.44(brd,J=15.1Hz,1H),8.39(d,J=4.8Hz,1H),7.45-7.56(m,1H),7.23 -7.35(m,4H),7.17-7.22(m,2H),4.94-5.11(m,2H),4.92(brs,1H),4.13- 4.33(m,2H),3.37-3.96(m,3H),3.04-3.25(m,1H),2.64-2.77(m,1H),1.9 4(s,3H),1.31-1.46(m,3H),1.07(d,J=6.6Hz,3H),0.94(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.03(s,1 F),-115.33(brd,J=13.0Hz,1 F),-125.45--124.18(m,2 F).m / z(ESI,+ve)695.2(M+H) + .

[0268] Example 12 (S)-6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methyl-4-(2-(2,3-difluorophenoxy)acetyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A mixture of (S)-6-chloro-4-(4-(2-chloroacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (51 mg, 0.09 mmol), 2,3-difluorophenol (34.1 mg, 0.26 mmol) and anhydrous potassium carbonate (60.4.0 mg, 0.44 mmol) in DMF (1 mL) was stirred and heated at 50° C. for 4 h. The resulting mixture was cooled to 0° C. and water (2 mL) was added. The resulting precipitate was collected by filtration, washed with water and dried to give (S)-6-chloro-4-(4-(2-(2,3-difluorophenoxy)acetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (38.2 mg, 0.06 mmol, 64.5% yield, 96% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.31-8.54(m,2H),7.41-7.58(m,1H),7.08-7.36(m,5H),6.88-7.06(m,2H),5.00-5.31(m,2H),4.93(brs,1H),4.10-4.38(m,2H),3.41- 4.01(m,3H),3.06-3.26(m,1H),2.64-2.74(m,1H),1.94(brs,3H),1.28-1.51(m,3H),1.07(brd,J=6.2Hz,3H),0.95(brd,J=6.4Hz,3H). 19 F NMR (376MHz, DMSO-d 6)δ ppm -114.01(brs,1 F),-138.60(brdd,J=20.8,10.4Hz,1 F),-160.51(brdd,J=53.8,20.8Hz,1 F).m / z(ESI,+ve)677.2(M+H) + .

[0269] Example 13 (S)-6-Chloro-4-(4-(2-diazoacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyrimidin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (210 mg, 0.35 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 30 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0270] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and 2-azidoacetic acid (52.4 mg, 0.05 mL, 0.52 mmol, Sigma-Aldrich Corporation) in DMF (1 mL) was added DIPEA (224 mg, 0.30 mL, 1.73 mmol, Sigma-Aldrich Corporation) followed by TBTU (167 mg, 0.52 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 3 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was diluted with MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-80% EtOAc / EtOH in heptane (3:1)) to give (S)-4-(4-(2-azidoacetyl)-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (198 mg, 0.34 mmol, 97% yield, 96% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.32-8.53(m,2H),7.46-7.59(m,1H),7.12-7.35(m,4H),4.82-5.03(m,1H),4.09-4.48(m,4H),3.51-3.89(m,3H),3.06-3.25(m,1H),2.6 5-2.76(m,1H),1.94(s,3H),1.38(brdd,J=6.74,11.92Hz,3H),1.07(d,J=6.63Hz,3H),0.95(d,J=6.84Hz,3H).m / z(ESI,+ve)590.2(M+H) + .

[0271] A solution of (S)-4-(4-(2-azidoacetyl)-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (94 mg, 0.16 mmol) in acetonitrile (2.0 mL) / water (0.2 mL) was treated with N-succinimidyl 3-(diphenylphosphino)propanoate (67.9 mg, 0.19 mmol, Aurum Pharmatech LLC) at 0° C. and stirred at room temperature for 4 h. m / z (ESI): 848.2 (M+H). + was observed. 1,8-Diazabicyclo-[5.4.0]undec-7-ene (DBU, 29.1 mg, 0.03 mL, 0.19 mmol, Sigma-Aldrich Corporation) was added and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with saturated NaCl solution (10 mL) and extracted with DCM (2 × 10 mL). The organic extracts were washed with MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-10% MeOH in DCM) to give (S)-6-chloro-4-(4-(2-diazoacetyl)-2-methylpiperazin-1-yl)-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (32 mg, 0.06 mmol, 34.9% yield, >95% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6)δ 8.44(s,1H),8.39(d,J= 4.8Hz,1H),7.55-7.47(m,1H),7.34-7.24(m,2H),7.23-7.16(m,2H),6.13(s,1H),4.98-4.84(m,1H),4.27(brd,J= 13.6Hz,1H),4.08(q,J= 5.2Hz,1H),3.74(brt,J= 10.9Hz,1H),3.49-3.35(m,1H),3.26-3.08(m,2H),2.76-2.67(m,1H),1.94(s,3H),1.36(d,J= 6.7Hz,3H),1.07(d,J= 6.7Hz,3H),0.94(d,J= 6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.02(s,1F).m / z(ESI):575.0(M+H) + .

[0272] Example 14 4-((S)-4-((R)aziridine-2-carbonyl)-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyrimidin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (106.8 mg, 0.18 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 15 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0273] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and (R)-1-tritylaziridine-2-carboxylic acid (87 mg, 0.26 mmol) in DMF (1 mL) was added DIPEA (0.15 mL, 0.90 mmol, Sigma-Aldrich Corporation) followed by TBTU (85 mg, 0.26 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 20 minutes. To the resulting mixture was added water (5 mL). The resulting precipitate was collected by filtration, washed with water and dried to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((R)-1-tritylaziridine-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a pale yellow solid. m / z (ESI, +ve) 817.4 (M+H). + .

[0274] A solution of 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((R)-1-tritylaziridine-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (144 mg, 0.18 mmol) was treated with triethylsilane (0.08 mL, 0.70 mmol) followed by trifluoroacetic acid (0.11 mL, 1.41 mmol) at 0° C. and stirred for 5 min. The reaction was allowed to go to completion. The reaction mixture was diluted with EtOAc (15 mL) and washed with brine (15 mL). The organic extract was washed with MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give 4-((S)-4-((R)-aziridine-2-carbonyl)-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (15 mg, 0.03 mmol, 14.8% yield, 97% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.47(s,1H),8.40(d,J=5.0Hz,1H),7.48-7.58(m,1H),7.16-7.37(m,5H),4.89-5.06(m,1H),4.01-4.43(m,4H),3.45- 4.00(m,4H),2.65-2.78(m,2H),1.95(s,3H),1.38(brd,J=6.4Hz,3H),1.08(d,J=6.6Hz,3H),0.96(brd,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.03(s,1 F).m / z(ESI,+ve)575.2(M+H) + .

[0275] Example 15 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((S)-oxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (100 mg, 0.17 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 30 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0276] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and potassium (S)-oxirane-2-carboxylate (31.2 mg, 0.25 mmol) in DMF (1 mL) was added DIPEA (0.14 mL, 0.78 mmol, Sigma-Aldrich Corporation) followed by TBTU (79 mg, 0.25 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was washed with saturated NH 4 Wash with Cl solution (15 mL) and MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((S)-oxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (48.8 mg, 0.09 mmol, 51.3% yield, 96% purity) as a yellow solid.1 H NMR (400MHz, DMSO-d 6 )δ 8.45(d,J=12.0Hz,1H),8.39(d,J=4.8Hz,1H),7.45-7.58(m,1H),7.24-7.36(m,2H) ),7.15-7.23(m,2H),4.96(brdJ=1.9Hz,1H),4.10-4.38(m,3H),3.93(dt,J=16.6, 3.2Hz,1H),3.48-3.86(m,2H),3.04-3.26(m,1H),2.80-3.03(m,2H),2.63-2.75(m ,1H),1.94(s,3H),1.29-1.48(m,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.02(s,1 F).m / z(ESI,+ve)577.3(M+H) + .

[0277] Example 16 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((R)-oxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (94 mg, 0.16 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 30 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0278] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and potassium (R)-oxirane-2-carboxylate (29.3 mg, 0.23 mmol) in DMF (1 mL) was added DIPEA (0.14 mL, 0.78 mmol, Sigma-Aldrich Corporation) followed by TBTU (75 mg, 0.23 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was washed with saturated NH 4 Cl (15 mL) solution and MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((R)-oxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (31.2 mg, 0.05 mmol, 34.9% yield, 96% purity) as a yellow solid.1 H NMR (400MHz, DMSO-d 6 )δ 8.46(d,J=6.0Hz,1H),8.40(d,J=4.8Hz,1H),7.48-7.58(m,1H),7.26-7.34(m,2H),7.1 6-7.25(m,2H),4.97(brd,J=3.7Hz,1H),4.15-4.37(m,2H),3.83-4.13(m,2H),3.45-3. 78(m,1H),3.05-3.28(m,2H),2.94-3.02(m,1H),2.79-2.88(m,1H),2.63-2.76(m,1H), 1.95(s,3H),1.38(dd,J=9.1,6.8Hz,3H),1.08(d,J=6.8Hz,3H),0.96(d,J=6.8Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.02(d,J=3.5Hz,1 F).m / z(ESI,+ve)577.3(M+H) + .

[0279] Example 17 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((2R,3S)-3-methyloxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((2S,3R)-3-methyloxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (100 mg, 0.17 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 30 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0280] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and Rac-(2R,3S)-3-methyl-2-otrans xylan carboxylic acid (33.6 mg, 0.33 mmol, Enamine) in DMF (1 mL) was added DIPEA (0.14 mL, 0.78 mmol, Sigma-Aldrich Corporation) followed by TBTU (106 mg, 0.33 mmol, Advanced ChemTech) at 0° C. and stirred at 0° C. for 30 min. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was washed with saturated NH 4 Wash with Cl (15 mL) and MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (7:3) / heptane) to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((2R,3S)-3-methyloxirane-2-carbonyl)piperazin-1-yl)pyrido[2, 3-d]pyrimidin-2(1H)-one and 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((2S,3R)-3-methyloxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (1:1 ratio, 78.5 mg, 0.13 mmol, 81% yield, 98% purity) were obtained. 1 H NMR (400MHz, DMSO-d 6 )δ 8.43-8.48(m,1H),8.40(d,J=5.02Hz,1H),7.48-7.58(m,1H),7.25-7.36(m,2H),7.18-7.24(m,2H),4.96(brs,1H),3.97-4.40(m,3H),3 .66-3.90(m,3H),3.00-3.26(m,2H),2.64-2.74(m,1H),1.95(s,3H),1.29-1.49(m,6H),1.08(d,J=6.69Hz,3H),0.95(d,J=6.69Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.03(s,1F).m / z(ESI,+ve)591.2(M+H) + .

[0281] Example 18 Ethyl (2S,3S)-3-((S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonyl)oxirane-2-carboxylate [ka] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (95 mg, 0.16 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 15 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0282] To a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and (2S,3S)-3-(ethoxycarbonyl)oxirane-2-carboxylic acid (50.1 mg, 0.31 mmol, Carbosynth Ltd.) in DMF (1 mL) was added DIPEA (0.14 mL, 0.78 mmol, Sigma-Aldrich Corporation) followed by TBTU (100 mg, 0.31 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was diluted with MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give ethyl (2S,3S)-3-((S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonyl)oxirane-2-carboxylate (12.2 mg, 0.02 mmol, 12% yield, 96% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.45(d,J=13.1Hz,1H),8.39(d,J=5.0Hz,1H),7.45-7.57(m,1H),7.16-7 .35(m,4H),4.95(brd,J=5.6Hz,1H),4.04-4.37(m,6H),3.49-3.88(m,3H ),3.06-3.27(m,1H),2.65-2.75(m,1H),1.94(d,J=2.5Hz,3H),1.33-1.4 6(m,3H),1.21-1.30(m,3H),1.07(d,J=6.4Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.03(d,J=1.73Hz,1F).m / z(ESI,+ve)649.2(M+H) + .

[0283] Examples 19 and 20 Ethyl (2R,3R)-3-((S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonyl)oxirane-2-carboxylate (19) and (2R,3R)-3-((S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonyl)oxirane-2-carboxylic acid (20). [ka] To an ice-cold solution of (2R,3R)-diethyl 2,3-epoxysuccinate (250 mg, 1.33 mmol, Sigma-Aldrich Corporation) in ethanol (3 mL) was added a solution of potassium hydroxide (89 mg, 1.594 mmol, VWR International, LLC) in ethanol (2.0 mL) dropwise over 2 min. After this addition, the reaction mixture was allowed to stir at 0 °C for 3 h. The resulting reaction mixture became a gel-like mixture. The cooling bath was then removed and the reaction mixture was further stirred at room temperature for 16 h. The ethanol was then removed in vacuo to leave an off-white solid. The salt was then taken up in 10 mL of water and washed three times with methane dichloride. The aqueous phase was acidified to pH 3 by the addition of 2 M HCl and extracted with ethyl acetate (10 mL x 3). The combined organic phase was diluted with MgSO 4 After drying, filtration and concentration in vacuo, the pure product was obtained after drying as a colorless oil (2R,3R)-3-(ethoxycarbonyl)oxirane-2-carboxylic acid (170 mg, 1.06 mmol, 80% yield). 1H NMR(400MHz,CDCl3)δ 8.41-9.03(brs,1H),4.29(ttd,J=3.6,7.1,10.7Hz,2H),3.72-3.73(m,1H),3.70-3.72(m,1H),1.33(t,J=7.2Hz,3H).[Reference: ACS Chemical Biology,5(3),279-285;2010].

[0284] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (105 mg, 0.17 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 15 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0285] To a mixture of TBTU (111 mg, 0.35 mmol, Advanced ChemTech) and (2R,3R)-3-(ethoxycarbonyl)oxirane-2-carboxylic acid (55.4 mg, 0.35 mmol) in DMF (1 mL) was added DIPEA (0.09 mL, 0.52 mmol, Sigma-Aldrich Corporation) at 0° C. and stirred for 5 min. The resulting mixture was added to a mixture of the above (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and DIPEA (0.09 mL, 0.52 mmol) in DMF (1 mL) at 0° C. and stirred at room temperature for 16 h. To the resulting mixture was added water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was washed with MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give ethyl (2R,3R)-3-((S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonyl)oxirane-2-carboxylate (39.5 mg, 0.06 mmol, 35.2% yield, 90% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6 )δ 8.45(d,J=8.1Hz,1H),8.39(d,J=5.0Hz,1H),7.46-7.56(m,1H),7.18-7.35(m ,4H),4.95(brd,J=2.5Hz,1H),4.13-4.37(m,6H),3.42-3.80(m,3H),3.05-3. 21(m,1H),2.63-2.78(m,1H),1.94(d,J=1.9Hz,3H),1.37(dd,J=6.8,9.7Hz,3 H),1.26(dt,J=3.5,7.1Hz,3H),1.07(d,J=6.6Hz,3H),0.95(d,J=6.6Hz,3H). 19 F NMR (377MHz, DMSO-d 6 )δ -114.03(s,1F).m / z(ESI,+ve)649.0(M+H) + .

[0286] The aqueous layer was acidified with 2N HCl to pH = 1-2 and extracted with DCM (10 mL x 2). The organic extracts were washed with MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-15% MeOH (with 2% AcOH) / DCM) to give (2R,3R)-3-((S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carbonyl)oxirane-2-carboxylic acid (6.5 mg, 10.5 μmol, 6% yield, 93% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6 )δ 11.92(s,1H),8.45(d,J=3.94Hz,1H),8.39(d,J=4.98Hz,1H),7.47-7.54(m,1H),7.24 -7.34(m,2H),7.16-7.23(m,2H),4.92-5.00(m,1H),4.27-4.35(m,2H),4.14-4.20(m, 1H),3.86-4.08(m,3H),3.69-3.78(m,1H),3.51(brs,1H),2.69-2.77(m,1H),1.91(s, 3H),1.37(brdd,J=6.74,14.82Hz,3H),1.07(d,J=6.63Hz,3H),0.95(d,J=6.63Hz,3H). 19 F NMR (377MHz, DMSO-d 6 )δ -114.03(s,1F).m / z(ESI,+ve)621.2(M+H) + .

[0287] Example 21 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((2R,3S)-3-(piperidin-1-ylmethyl)oxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] To an ice-cold solution of (2R,3R)-diethyl 2,3-epoxysuccinate (900 mg, 4.78 mmol, Sigma-Aldrich Corporation) in ethanol (3 mL) was added sodium borohydride (145 mg, 3.83 mmol, Sigma-Aldrich Corporation). After this addition, the reaction mixture was allowed to stir in an ice bath for 2 h. The reaction mixture was slowly quenched with water (1.0 mL) and then acidified to pH 6-7 by addition of 2 M aqueous HCl. The ethanol was then removed under reduced pressure to leave an off-white solid. The salt was then taken up in 15 mL water and washed three times with methane dichloride. The aqueous phase was acidified to pH 3 by addition of concentrated hydrochloric acid and extracted four times with DCM. The combined organic phases were dried over magnesium sulfate and concentrated under reduced pressure to give ethyl (2R,3S)-3-(hydroxymethyl)oxirane-2-carboxylate (413.8 mg, 2.83 mmol, 59.2% yield) as a yellow oil. 1 H NMR (400 MHz, CDCl 3 )δ 4.18-4.33(m,2H),4.01(ddd,J=2.18,5.08,13.06Hz,1H),3.72-3.81(m,1H),3.54(d,J=2.07Hz,1H),3.39(td,J= 2.10,3.27Hz,1H),1.64(dd,J=5.18,8.09Hz,1H),1.31(t,J=7.15Hz,3H).(Reference: J.Org.Chem.2005,70,5643-5654).

[0288] To an ice-cold solution of ethyl (2R,3S)-3-(hydroxymethyl)oxirane-2-carboxylate (410 mg, 2.81 mmol) in dichloromethane (12 mL) was added triethylamine (0.59 mL, 4.21 mmol, Sigma-Aldrich Corporation). After 5 min, the reaction mixture was treated with methanesulfonyl chloride (0.33 mL, 4.21 mmol, Sigma-Aldrich Corporation) and stirred at 0° C. for 10 min and at room temperature for 2 h. The solvent was then evaporated. The resulting yellow paste was dissolved in N,N-dimethylformamide (12.00 mL) and potassium iodide (23.3 mg, 0.14 mmol, Fisher Scientific UK) was added. The yellow solution was cooled to 0° C. before piperidine (0.83 mL, 8.42 mmol, Spectrum Chemicals & Laboratory Products) was added dropwise. The resulting suspension was allowed to warm slowly to room temperature and stirred for 24 h. The resulting reaction mixture was diluted with saturated NaHCO 3 The organic extract was diluted with aqueous solution (10 mL) and extracted with DCM (20 mL x 2). 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-4% MeOH / DCM) to give ethyl (2R,3S)-3-(piperidin-1-ylmethyl)oxirane-2-carboxylate (90.2 mg, 0.423 mmol, 15.07% yield) as a brown oil. 1 H NMR (400 MHz, CDCl 3 )δ 4.19-4.33(m,2H),3.35-3.44(m,1H),3.24(d,J=1.87Hz,1H),2.82(brs,1H),2.43-2.68(m,4H),2.38( dd,J=6.63,13.68Hz,1H),1.64(brs,4H),1.46(brs,2H),1.30(t,J=7.15Hz,3H).m / z(ESI):214.2(M+H) + (Reference: J.Med.Chem.2010,53,2038-2050).

[0289] To an ice-cold solution of ethyl (2R,3S)-3-(piperidin-1-ylmethyl)oxirane-2-carboxylate (90 mg, 0.422 mmol) in ethanol (1.5 mL) was added a solution of potassium hydroxide (28.4 mg, 0.506 mmol, VWR International, LLC) in ethanol (1.5 mL) dropwise over 2 min. After this addition, the reaction mixture was allowed to stir in ice for 2 h and at room temperature for 17 h. The resulting reaction mixture became a brown solution. The ethanol was then removed under reduced pressure leaving a brown oil which was used in the next step without purification. m / z(ESI): 186.2 (M+H) + .

[0290] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (93 mg, 0.15 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL, 13.4 mmol) at room temperature and stirred for 15 min. The reaction mixture was concentrated in vacuo to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0291] DIPEA (0.13 mL, 0.77 mmol, Sigma-Aldrich Corporation) was added to a mixture of TBTU (98 mg, 0.31 mmol, Advanced ChemTech) and (2R,3S)-3-(piperidin-1-ylmethyl)oxirane-2-carboxylic acid (78 mg, 0.42 mmol) in N,N-dimethylformamide (1 mL) at 0° C. and stirred for 5 min. The above mixture was added to a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and DIPEA (0.13 mL, 0.77 mmol) in N,N-dimethylformamide (1 mL) at 0° C. The reaction mixture was stirred at room temperature for 3 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (20 mL x 2). The organic extracts were washed with MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-10% MeOH / DCM) to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((S)-2-methyl-4-((2R,3S)-3-(piperidin-1-ylmethyl)oxirane-2-carbonyl)piperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (56.3 mg, 0.08 mmol, 54.5% yield, >99% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6)δ 8.45(d,J=7.11Hz,1H),8.39(d,J=4.81Hz,1H),7.47-7.55(m,1H),7.24-7.34(m,2H), 7.17-7.24(m,2H),4.90-5.02(m,1H),3.96-4.36(m,3H),3.36-3.94(m,4H),3.02-3.2 4(m,2H),2.68-2.80(m,2H),2.36-2.46(m,3H),2.23-2.32(m,1H),1.94(s,3H),1.52( brd,J=2.30Hz,4H),1.30-1.42(m,5H),1.07(d,J=6.69Hz,3H),0.95(d,J=6.69Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.02(d,J=4.34Hz,1F).m / z(ESI,+ve)674.2(M+H) + .

[0292] Example 22 6-Chloro-7-(2-fluorophenyl)-4-((S)-4-((2R,3S)-3-(hydroxymethyl)oxirane-2-carbonyl)-2-methylpiperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] To a mixture of ethyl (2R,3S)-3-(hydroxymethyl)oxirane-2-carboxylate (300 mg, 2.05 mmol) and imidazole (280 mg, 4.11 mmol, Sigma-Aldrich Corporation) in tetrahydrofuran (4 mL) was added (1,1-dimethylethyl)dimethylsilyl chloride (309 mg, 2.053 mmol, Combi-Blocks Inc.). After this addition, the reaction mixture was allowed to stir at room temperature for 1 h. The reaction mixture was quenched with water (5 mL) and extracted with DCM (10 mL x 4). The combined organic phase was diluted with MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-10% EtOAc / heptane) to give ethyl (2R,3S)-3-(((tert-butyldimethylsilyl)oxy)methyl)oxirane-2-carboxylate (415.1 mg, 1.59 mmol, 78% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl 3 )δ 4.09-4.25(m,2H),3.80-3.89(m,1H),3.68-3.75(m,1H),3.36(d,J=1.87Hz,1H),3.25(td,J=2.33 ,3.42Hz,1H),1.24(t,J=7.15Hz,3H),0.83(s,9H),0.00(d,J=3.32Hz,6H).m / z(ESI):261.2(M+H) + .

[0293] To an ice-cold solution of ethyl (2R,3S)-3-(((tert-butyldimethylsilyl)oxy)methyl)oxirane-2-carboxylate (410 mg, 1.57 mmol) in ethanol (1.5 mL) was added a solution of potassium hydroxide (106 mg, 1.89 mmol, VWR International, LLC) in ethanol (1.5 mL) dropwise over 2 min. After this addition, the reaction mixture was allowed to stir in the ice bath for 5 min and at room temperature for 24 h. The resulting reaction mixture became a brown solution. The ethanol was then removed under reduced pressure to give potassium (2R,3S)-3-(((tert-butyldimethylsilyl)oxy)methyl)oxirane-2-carboxylate as a pale yellow solid, which was used in the next step without purification. m / z (ESI): 186.2 (M+H). + .

[0294] A solution of tert-butyl (S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-3-methylpiperazine-1-carboxylate (103 mg, 0.17 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1.0 mL, 13.4 mmol) at room temperature and stirred for 15 minutes. The reaction was completed and concentrated to give (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 507.2 (M+H). + .

[0295] To a mixture of TBTU (109 mg, 0.34 mmol, Advanced ChemTech) and potassium (2R,3S)-3-(((tert-butyldimethylsilyl)oxy)methyl)oxirane-2-carboxylate (138 mg, 0.51 mmol) in N,N-dimethylformamide (1 mL) was added DIPEA (0.15 mL, 0.85 mmol) at 0° C. and stirred for 5 min. The above mixture was added to a mixture of (S)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-methylpiperazin-1-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and DIPEA (0.15 mL, 0.85 mmol) in N,N-dimethylformamide (1 mL) at 0° C. The reaction mixture was stirred at room temperature for 30 min. The resulting mixture was added with water (5 mL) and extracted with EtOAc (20 mL x 2). The organic extracts were washed with MgSO 4The solution was filtered, concentrated in vacuo and purified by silica gel chromatography (eluent: 0-10% MeOH / DCM) to give 4-((S)-4-((2R,3S)-3-(((tert-butyldimethylsilyl)oxy)methyl)oxirane-2-carbonyl)-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (101 mg, 0.14 mmol, 83% yield) as a pale yellow solid. m / z (ESI): 721.2 (M+H). + .

[0296] 4-((S)-4-((2R,3S)-3-(((tert-butyldimethylsilyl)oxy)methyl)oxirane-2-carbonyl)-2-methylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (101 mg, 0.14 mmol) was dissolved in THF (1.0 mL) and treated with TBAF (tetrabutylammonium fluoride solution, 1.0 M in tetrahydrofuran, 0.34 mL, 0.34 mmol, Sigma-Aldrich Corporation) at 0° C. and stirred at room temperature for 5 h. The reaction mixture was diluted with water (10 mL) and extracted with DCM (3×10 mL). The organic extract was washed with MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-10% MeOH / DCM) to give 6-chloro-7-(2-fluorophenyl)-4-((S)-4-((2R,3S)-3-(hydroxymethyl)oxirane-2-carbonyl)-2-methylpiperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (8.1 mg, 0.01 mmol, 7.9% yield, 99% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6)δ 8.45(d,J=6.06Hz,1H),8.39(d,J=5.02Hz,1H),7.47-7.57(m,1H),7.24-7.35(m,2H),7.17-7.24( m,2H),4.97-5.07(m,1H),4.89-4.97(m,1H),4.24-4.38(m,2H),4.18(brd,J=13.17Hz,1H),3.90-4 .04(m,1H),3.80(d,J=2.09Hz,1H),3.65-3.76(m,2H),3.49-3.58(m,1H),3.03-3.26(m,2H),2.68 -2.77(m,1H),1.94(s,3H),1.37(t,J=7.11Hz,3H),1.07(d,J=6.69Hz,3H),0.95(d,J=6.69Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.02(s,1F).m / z(ESI):607.2(M+H) + .

[0297] Example 23 2-((2S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-1-(oxirane-2-carbonyl)piperazin-2-yl)acetonitrile [ka] A mixture of 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (WO 2019051291A1) (200 mg, 0.47 mmol) and DIPEA (0.12 mL, 0.71 mmol) in acetonitrile (10 mL) was added with phosphorus oxychloride (0.07 mL, 0.71 mmol, Sigma-Aldrich Corporation) at room temperature and stirred at 80 ° C for 30 minutes. The reaction mixture was concentrated to give 4,6-dichloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a brown solid. m / z (ESI): 443.0 (M + H) + .

[0298] To a mixture of 4,6-dichloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and DIPEA (0.24 mL, 1.42 mmol) in acetonitrile (10 mL) was added (S)-2-(piperazin-2-yl)acetonitrile hydrochloride (152 mg, 0.94 mmol, AstaTech Inc.) and stirred at room temperature for 1.5 h. The crude reaction mixture was concentrated and purified by silica gel chromatography (eluent: 0-10% MeOH (2N NH 3 Purification by hexanes / DCM) gave (S)-2-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (250 mg, 96% purity) as a yellow foam. m / z (ESI, +ve) 532.1 (M+H).

[0299] To a mixture of (S)-2-(4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)piperazin-2-yl)acetonitrile (250 mg, 0.470 mmol) and potassium 2-oxiranecarboxylate (89 mg, 0.71 mmol, Enamine) in DMF (2 mL) was added DIPEA (121 mg, 0.16 mL, 0.94 mmol) followed by TBTU (226 mg, 0.71 mmol, Advanced ChemTech) at 0 °C. The reaction mixture was stirred at room temperature for 18 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL x 2). The organic extract was washed with MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-50% EtOAc / EtOH (3:1) / heptane) to give 2-((2S)-4-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-1-(oxirane-2-carbonyl)piperazin-2-yl)acetonitrile (77.6 mg, 0.13 mmol, 27.4% yield, 95% purity) as a pale yellow solid. 1 H NMR (400MHz, DMSO-d 6 ,)δ 8.63(s,1H),8.40(d,1H,J=5.0Hz),7.5-7.6(m,1H),7.3-7.3(m,2H),7.2-7.2(m,2H),4.8-5.1(m,1H),4.1-4.5(m,3H),3.9-4.0(m,1H),3. 7-3.9(m,1H),3.5-3.7(m,1H),3.4-3.4(m,1H),3.0-3.2(m,3H),2.7-2.9(m,2H),1.92(s,3H),1.07(d,3H,J=6.8Hz),0.93(d,3H,J=6.6Hz). 19 F NMR (DMSO-d 6 ,376MHz)δ -114.4--114.1(m,1F).m / z(ESI):602.2(M+H) + .

[0300] Example 24 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(6-(oxirane-2-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] To a mixture of 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione (WO2019051291A1) (272 mg, 0.64 mmol) and DIPEA (0.12 mL, 0.96 mmol) in acetonitrile (3 mL), phosphorus oxychloride (0.09 mL, 0.96 mmol, Sigma-Aldrich Corporation) was added at room temperature and then stirred at 80° C. for 1 h. The reaction mixture was concentrated to give 4,6-dichloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one as a brown solid. m / z (ESI): 443.0 (M+H) + .

[0301] To a mixture of 4,6-dichloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and DIPEA (0.17 mL, 0.96 mmol) in acetonitrile (3 mL) was added 2-boc-2,6-diazaspiro[3.3]heptane (210 mg, 1.06 mmol, AstaTech, Inc) and stirred at room temperature for 10 minutes. The crude reaction mixture was concentrated and purified by silica gel chromatography (eluent: 0-60% EtOAc / EtOH (3:1) / heptane) to give tert-butyl 6-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (141 mg, 36.4% yield, 88% purity) as a pale yellow foam. 1 H NMR (400MHz, DMSO-d 6 ,)δ 8.36(d,J=4.8Hz,1H),8.27(s,1H),7.4-7.6(m,1H),7.2-7.3(m,2H),7.1-7.2(m,2H),5.09(brs,2H),4.48(br s,2H),4.12(brs,4H),2.6-2.7(m,1H),1.91(s,3H),1.40(s,9H),1.05(d,J=6.7Hz,3H),0.91(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.56(s,1F).m / z(ESI,+ve)605.2(M+H) + .

[0302] A solution of tert-butyl 6-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (140 mg, 0.23 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1.0 mL) at room temperature and stirred for 30 minutes. The reaction was completed and concentrated to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2,6-diazaspiro[3.3]heptane-2-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 505.2 (M+H). + .

[0303] To a mixture of the above 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2,6-diazaspiro[3.3]heptan-2-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and potassium 2-oxiranecarboxylate (58.4 mg, 0.46 mmol, Enamine) in DMF (1 mL) was added DIPEA (0.20 mL, 1.16 mmol) followed by TBTU (149 mg, 0.46 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 2 h. The resulting mixture was added with water (5 mL) and extracted with EtOAc (10 mL×2). The organic extract was diluted with MgSO 4 The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-50% EtOAc / EtOH (3:1) / heptane) to give 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(6-(oxirane-2-carbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (6 mg, 10.4 μmol, 4.51% yield, 96% purity) as a white solid. 1 H NMR (400MHz, DMSO-d 6)δ 8.37(d,J=4.8Hz,1H),8.28(s,1H),7.5-7.5(m,1H),7.3-7.3(m,2H),7.2-7.3(m,1H),7. 17(dd,J=0.7,4.9Hz,1H),5.12(brd,J=2.3Hz,2H),4.59(brs,2H),4.52(brs,2H),4.20(b rd,J=1.7Hz,2H),3.48(dd,J=2.6,4.3Hz,1H),2.91(dd,J=4.4,6.5Hz,1H),2.82(dd,J=2. 5,6.5Hz,1H),2.6-2.7(m,1H),1.91(s,3H),1.05(d,J=6.7Hz,3H),0.92(d,J=6.7Hz,3H). 19 F NMR (DMSO-d 6 ,376MHz)δ -114.57(s,1F).m / z(ESI):575.0(M+H) + .

[0304] Example 25 (R)-6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(2-(oxirane-2-carbonyl)-2,7-diazaspiro[3.5]nonan-7-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] The title compound (67.3 mg, 0.11 mmol, 47% yield, 97% purity, pale yellow solid) was prepared in a manner similar to compound 24 by replacing 2-boc-2,6-diazaspiro[3.3]heptane with 2-(tert-butoxycarbonyl)-2,7-diazaspiro[3.5]nonane (Combi-Blocks Inc.) in step 2 and potassium 2-oxiranecarboxylate with potassium (R)-oxirane-2-carboxylate (Aurum Pharmatech LLC) in step 4. 1 H NMR (400MHz, DMSO-d 6)δ 8.42(s,1H),8.39(d,J=5.02Hz,1H),7.47-7.57(m,1H),7.25-7.35(m,2H),7.18 -7.25(m,2H),4.11-4.21(m,2H),3.87(brs,4H),3.77(s,2H),3.49(dd,J=2.51,4 .18Hz,1H),2.89-2.94(m,1H),2.84(dd,J=2.51,6.27Hz,1H),2.67-2.75(m,1H) ,1.96-2.00(m,4H),1.93(s,3H),1.07(d,J=6.90Hz,3H),0.95(d,J=6.69Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.02(d,J=4.34Hz,1F).m / z(ESI):603.2(M+H) + .

[0305] Example 26 N-((R)-1-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)pyrrolidin-3-yl)oxirane-2-carboxamide [ka] The title compound (122.7 mg, 0.22 mmol, 59.6% yield, 96% purity, pale yellow solid) was prepared in a manner analogous to compound 24 by substituting (R)-3-(boc-amino)pyrrolidine (Ontario Chemical, Inc.) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6)δ 8.64(brs,1H),8.50(brd,J=6.3Hz,1H),8.37(d,J=4.8Hz,1H),7.5-7.5(m,1H),7.2-7.3(m,2H) ),7.2-7.2(m,2H),4.46(brs,1H),3.8-4.2(m,3H),3.4-3.5(m,1H),3.39(dd,J=2.5,4.2Hz,1H) ,2.91(dd,J=4.4,6.3Hz,1H),2.83(dd,J=2.5,6.3Hz,1H),2.7-2.8(m,1H),2.21(brd,J=3.3Hz ,1H),2.06(brd,J=5.4Hz,1H),1.91(s,3H),1.07(dd,J=0.7,6.8Hz,3H),0.92(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.8--114.6(m,1F).m / z(ESI):563.0(M+H) + .

[0306] Example 27 N-((R)-1-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)pyrrolidin-3-yl)oxirane-2-carboxamide [ka] The title compound (88.4 mg, 0.16 mmol, 42.8% yield, 96% purity, pale yellow solid) was prepared in a manner similar to compound 24 by substituting 3-N-boc-amino-azetidine (AstaTech, Inc) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6)δ 8.85(brd,J=6.5Hz,1H),8.37(d,J=4.8Hz,1H),8.30(s,1H),7.5-7.6(m,1H),7.2-7.3(m,2H) ),7.2-7.2(m,2H),5.19(brdd,J=2.0,7.8Hz,1H),4.7-4.8(m,2H),4.59(brdd,J=2.7,6.1Hz ,1H),4.2-4.4(m,1H),3.44(dd,J=2.5,4.4Hz,1H),2.97(dd,J=4.5,6.2Hz,1H),2.85(dd,J= 2.5,6.1Hz,1H),2.6-2.7(m,1H),1.92(s,3H),1.06(d,J=6.7Hz,3H),0.92(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ-114.55(s,1F).m / z(ESI):549.2(M+H) + .

[0307] Example 28 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(6-(oxirane-2-carbonyl)-2,6-diazabicyclo[3.2.0]heptan-2-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] The title compound (12.4 mg, 0.02 mmol, 16.1% yield, 93% purity, white solid) was prepared in a manner similar to compound 24 by substituting 6-boc-2,6-diazabicyclo[3.2.0]heptane (Aurum Pharmatech LLC) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6)δ8.55(brs,1H),8.38(d,J=4.8Hz,1H),7.5-7.6(m,1H),7.2-7.3(m,2H),7.2 -7.2(m,2H),5.38(brs,1H),5.29(brd,J=2.7Hz,1H),5.0-5.1(m,1H),4.5-4. 8(m,2H),4.2-4.4(m,2H),3.6-3.7(m,1H),3.4-3.5(m,1H),2.8-3.0(m,2H),2 .7-2.7(m,1H),1.93(d,J=8.2Hz,3H),1.0-1.1(m,3H),0.92(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.7--114.0(m,1F)m / z(ESI):575.1(M+H) + .

[0308] Example 29 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(methyl(1-(oxirane- [ka] The title compound (12.4 mg, 0.02 mmol, 16.1% yield, 93% purity, white solid) was prepared in a manner similar to compound 24 by substituting tert-butyl 3-(methylamino)azetidine-1-carboxylate (AstaTech, Inc) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6 )δ8.55(brd,J=2.3Hz,1H),8.40(brd,J=4.2Hz,1H),7.4-7.5(m,1H),7.2-7.3(m,4H),4.6-4.8(m,1H),3.9-4.5(m,1H) ),3.6-3.9(m,4H),3.11(brs,2H),2.8-3.0(m,3H),2.8-2.8(m,1H),2.0-2.1(m,3H),1.0-1.1(m,3H),0.9-1.0(m,3H). 19 F NMR (376MHz, DMSO-d 6)δ-114.9--114.5(m,1F).m / z(ESI):563.0(M+H) + .

[0309] Example 30 (R)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(methyl(1-(oxirane-2-carbonyl)azetidin-3-yl)amino)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] The title compound (12.4 mg, 0.02 mmol, 16.1% yield, 93% purity, white solid) was prepared in a manner similar to compound 24 by replacing 2-boc-2,6-diazaspiro[3.3]heptane with tert-butyl 3-(methylamino)azetidine-1-carboxylate (AstaTech, Inc) in step 2 and potassium 2-oxiranecarboxylate with potassium (R)-oxirane-2-carboxylate (Aurum Pharmatech LLC) in step 4. 1 H NMR (400MHz, DMSO-d 6 )δ8.55(s,1H),8.40(d,J=4.8Hz,1H),7.4-7.5(m,1H),7.1-7.3(m,4H),4.5-4.8(m,1H),4.0-4.3(m,1H),3.6-4.0(m,4H) ,3.11(d,J=4.0Hz,2H),2.8-3.0(m,3H),2.80(brdd,J=3.3,6.1Hz,1H),2.0-2.1(m,3H),1.1-1.1(m,3H),0.9-1.0(m,3H). 19 F NMR (376MHz, DMSO-d 6 )δ-114.03(d,J=1.73Hz,1F).m / z(ESI):563.2(M+H) + .

[0310] Example 31 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-((1-(oxirane-2-carbonyl)azetidin-3-yl)amino)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] The title compound (30.5 mg, 0.06 mmol, 24.7% yield, 97% purity, white solid) was prepared in a manner analogous to compound 24 by substituting tert-butyl 3-aminoazetidine-1-carboxylate (Enamine) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6 )δ 9.31(brs,1H),8.99(d,J=3.6Hz,1H),8.37(d,J=4.8Hz,1H),7.4-7.6(m,1H), 7.2-7.3(m,2H),7.2-7.2(m,1H),7.2-7.2(m,1H),4.9-5.1(m,1H),4.7-4.8(m, 1H),4.3-4.4(m,2H),4.0-4.2(m,1H),3.5-3.6(m,1H),2.9-3.0(m,1H),2.8-2. 9(m,1H),2.7-2.7(m,1H),1.92(s,3H),1.0-1.1(m,3H),0.92(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.68(s,1F).m / z(ESI):549.2(M+H) + .

[0311] Example 32 6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(((3R)-1-(oxirane-2-carbonyl)pyrrolidin-3-yl)amino)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] The title compound (82.7 mg, 0.15 mmol, 30% yield, 96% purity, white solid) was prepared in a manner similar to compound 24 by replacing 2-boc-2,6-diazaspiro[3.3]heptane with (R)-(+)-N-boc-3-aminopyrrolidine (CNH Technologies, Inc.) in step 2. 1 H NMR (400MHz, DMSO-d 6 )δ 9.0-9.1(m,1H),8.7-8.9(m,1H),8.37(d,J=4.8Hz,1H),7.5-7.5(m,1H) ,7.2-7.3(m,2H),7.1-7.2(m,2H),4.7-5.0(m,1H),3.8-4.2(m,2H),3.5 -3.8(m,3H),2.9-3.0(m,1H),2.8-2.9(m,1H),2.7-2.8(m,1H),2.0-2.4 (m,2H),1.93(s,3H),1.06(d,J=6.7Hz,3H),0.92(dd,J=2.5,6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -114.8--114.6(m,1F).m / z(ESI):563.2(M+H) + .

[0312] Example 33 6-Chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-4-(7-(oxirane-2-carbonyl)-2,7-diazabicyclo[4.2.0]octan-2-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] The title compound (91.8 mg, 0.16 mmol, 77% yield, 96% purity, pale yellow solid) was prepared in a manner similar to compound 24 by substituting 7-boc-2,7-diazabicyclo[4.2.0]octane (Aurum Pharmatech LLC) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6)δ 8.56(brs,1H),8.40(d,J=4.8Hz,1H),7.4-7.6(m,1H),7.2-7.3(m,2H), 7.2-7.2(m,2H),4.86(ddd,J=2.5,7.8,10.3Hz,1H),3.6-4.5(m,6H),2.9 -3.0(m,2H),2.8-2.9(m,1H),2.1-2.2(m,1H),2.04(d,J=8.1Hz,3H),1. 82(brd,J=1.9Hz,1H),1.4-1.7(m,2H),1.1-1.1(m,3H),0.9-1.0(m,3H). 19 F NMR (376MHz, DMSO-d 6 )δ-115.0--114.5(m,1F).m / z(ESI):589.1(M+H) + .

[0313] Example 34 N-((S)-1-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)piperidin-3-yl)oxirane-2-carboxamide [ka] The title compound (168 mg, 0.29 mmol, 70.8% yield, 95% purity, pale yellow solid) was prepared in a manner similar to compound 24 by substituting (S)-3-boc-aminopiperidine (Aurum Pharmatech LLC) for 2-boc-2,6-diazaspiro[3.3]heptane in step 2. 1 H NMR (400MHz, DMSO-d 6)δ8.53(d,J=2.1Hz,1H),8.38(d,J=4.8Hz,1H),8.24-8.37(m,1H),7.46-7.55(m,1H),7 .24-7.34(m,2H),7.16-7.23(m,2H),4.13-4.34(m,2H),3.95-4.02(m,1H),3.33-3.52(m ,3H),2.88(ddd,J=13.1,6.3,4.4Hz,1H),2.72-2.83(m,1H),2.59-2.69(m,1H),1.97-2. 02(m,2H),1.94(s,3H),1.61-1.86(m,2H),1.06(d,J=6.7Hz,3H),0.94(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ-114.09(d,J=12.1Hz,1 F).m / z(ESI):577.2(M+H) + .

[0314] Example 35 N-((S)-1-(6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)pyrrolidin-3-yl)oxirane-2-carboxamide [ka] The title compound (117 mg, 0.21 mmol, 48.4% yield, 95% purity, pale yellow solid) was prepared in a manner similar to compound 24 by replacing 2-boc-2,6-diazaspiro[3.3]heptane with (S)-(−)-3-(boc-amino)pyrrolidine (CNH Technologies, Inc.) in step 2. 1 H NMR (400MHz, DMSO-d 6)δ8.64(brs,1H),8.48(brd,J=6.7Hz,1H),8.37(d,J=4.8Hz,1H),7.5-7.5(m,1H),7.2-7.3(m, 2H),7.2-7.2(m,2H),4.46(brd,J=4.2Hz,1H),3.7-4.1(m,3H),3.4-3.5(m,1H),3.39(dd,J=2. 4,4.3Hz,1H),2.91(dd,J=4.4,6.3Hz,1H),2.83(ddd,J=1.1,2.4,6.3Hz,1H),2.6-2.7(m,1H), 2.1-2.3(m,1H),2.04(brd,J=5.2Hz,1H),1.94(s,3H),1.0-1.1(m,3H),0.92(d,J=6.7Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ-114.30(s,1F).m / z(ESI):563.2(M+H) + .

[0315] Example 36 6-Chloro-4-((2S,5R)-2,5-dimethyl-4-((R)-oxirane-2-carbonyl)piperazin-1-yl)-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (2R,5S)-4-(6-chloro-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (WO2019051291A1) (140 mg, 0.22 mmol) in dichloromethane (1 mL) was treated with TFA (1.0 mL) at room temperature and stirred for 10 min. The reaction was completed and concentrated to give 6-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 537.2 (M+H). + .

[0316] To a mixture of 6-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and potassium (R)-oxirane-2-carboxylate (41.6 mg, 0.33 mmol, Aurum Pharmatech LLC) in N,N-dimethylformamide (1 mL) was added DIPEA (0.19 mL, 1.10 mmol) followed by TBTU (106 mg, 0.33 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The resulting mixture was added with water (10 mL) and extracted with EtOAc (20 mL×2). The organic extract was purified by filtration using MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give 6-chloro-4-((2S,5R)-2,5-dimethyl-4-((R)-oxirane-2-carbonyl)piperazin-1-yl)-7-(2-fluoro-6-hydroxyphenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (39.5 mg, 0.07 mmol, 29.6% yield, 96% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ 10.01-10.24(m,1H),8.43(brs,1H),8.38(d,J=4.77Hz,1H),7.20-7.28(m,1H),7.18(d,J=4.7 7Hz,1H),6.62-6.74(m,2H),4.86(brd,J=1.24Hz,1H),4.57-4.74(m,1H),4.04-4.32(m,2H),3. 79-3.95(m,3H),2.93-3.01(m,1H),2.83(dd,J=2.38,6.32Hz,1H),2.65-2.76(m,1H),1.91(br s,3H),1.29-1.39(m,4H),1.19-1.29(m,2H),1.07(d,J=6.84Hz,3H),0.94(brd,J=6.43Hz,3H). 19 F NMR (376MHz, DMSO-d 6 )δ -115.83(brd,J=267.89Hz,1F).m / z(ESI,+ve)607.2(M+H) + .

[0317] Example 37 7-(2-amino-6-fluorophenyl)-6-chloro-4-((2S,5R)-2,5-dimethyl-4-((R)-oxirane-2-carbonyl)piperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one [ka] A solution of tert-butyl (2R,5S)-4-(7-(2-amino-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydropyrido[2,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-1-carboxylate (WO2019051291A1) (260 mg, 0.41 mmol) in dichloromethane (1 mL) was treated with trifluoroacetic acid (1 mL) at room temperature and stirred for 10 minutes. The reaction was completed and concentrated to give 7-(2-amino-6-fluorophenyl)-6-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one. m / z (ESI, +ve) 536.2 (M+H). + .

[0318] To a mixture of 7-(2-amino-6-fluorophenyl)-6-chloro-4-((2S,5R)-2,5-dimethylpiperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one and potassium (R)-oxirane-2-carboxylate (77 mg, 0.613 mmol, Aurum Pharmatech LLC) in N,N-dimethylformamide (1 mL) was added DIPEA (0.36 mL, 2.04 mmol) followed by TBTU (197 mg, 0.61 mmol, Advanced ChemTech) at 0° C. The reaction mixture was stirred at room temperature for 1 h. The resulting reaction mixture was added with water (10 mL) and extracted with EtOAc (20 mL×2). The organic extract was diluted with MgSO 4The solution was filtered, concentrated in vacuo, and purified by silica gel chromatography (eluent: 0-100% EtOAc-MeOH (9:1) / heptane) to give 7-(2-amino-6-fluorophenyl)-6-chloro-4-((2S,5R)-2,5-dimethyl-4-((R)-oxirane-2-carbonyl)piperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)pyrido[2,3-d]pyrimidin-2(1H)-one (173 mg, 0.29 mmol, 69.9% yield, 99% purity) as a yellow solid. 1 H NMR (400MHz, DMSO-d 6 )δ ppm 8.44(s,1H),8.39(d,J=4.8Hz,1H),7.19(d,J=4.8Hz,1H),7.05(q,J=7.5Hz,1H), 6.45(d,J=8.3Hz,1H),6.26-6.38(m,1H),5.06-5.16(m,2H),4.80-4.92(m,1H),4. 56-4.77(m,1H),4.01-4.22(m,2H),3.86-3.99(m,2H),2.97(dd,J=6.0,4.6Hz,1H) ,2.79-2.88(m,1H),2.57-2.73(m,1H),1.81-2.03(m,3H),1.23-1.41(m,5H),1.14 -1.21(m,2H),0.87-1.10(m,6H). 19 F NMR (376MHz, DMSO-d 6 )δ ppm -115.83--115.48(m,1 F).m / z(ESI,+ve)606.2(M+H) + .

[0319] Table 1: Biochemical and cellular activities of compounds For the compounds in Table 1, the following assay conditions were used:

[0320] Binding Nucleotide Exchange Assay: Purified GDP-bound KRAS protein (amino acids 1-169) containing either G12C or G12D and C118A amino acid substitutions and an N-terminal His tag was incubated for either 5 min or 20 h in assay buffer (25 mM HEPES (pH 7.4), 10 mM MgCl 2 and 0.01% Triton X-100) (see Table 15). After compound preincubation, purified SOS protein (amino acids 564-1049) and GTP (Roche 10106399001) were added to the assay wells and incubated for an additional 30 minutes. To determine the extent of inhibition of SOS-mediated nucleotide exchange, purified GST-tagged cRAF (amino acids 1-149), nickel-chelating AlphaLISA acceptor beads (PerkinElmer AL108R) and AlphaScreen glutathione donor beads (PerkinElmer 6765302) were added to the assay wells and incubated for 10 minutes. The assay plates were then read using AlphaScreen® technology on a PerkinElmer EnVision Multilabel Reader and the data were analyzed using a 4-parameter logistic model to determine IC 50 The value was calculated.

[0321] Phospho-ERK1 / 2 MSD Assay: MIA PaCa-2 (ATCC® CRL-1420™) cells were cultured in RPMI 1640 medium (ThermoFisher Scientific 11875093) containing 10% fetal bovine serum (ThermoFisher Scientific 16000044) and 1× penicillin-streptomycin-glutamine (ThermoFisher Scientific 10378016). 16 hours prior to compound treatment, MIA PaCa-2 cells were seeded into 96-well cell culture plates at a density of 25,000 cells / well and incubated at 37° C., 5% CO 2Dose-response titrations of compounds were diluted in growth medium and added to appropriate wells of the cell culture plate, followed by incubation at 37°C and 5% CO. 2 After compound treatment, cells were incubated with ice-cold Dulbecco's phosphate buffered saline (Ca 2+ or Mg 2+ Cells were washed with 100 mM Tris-HCl (pH 7.5), 1% Igepal, 0.5% sodium deoxycholate, 150 mM NaCl, and 0.5% sodium dodecyl sulfate containing protease inhibitors (Roche 4693132001) and phosphatase inhibitors (Roche 4906837001) and then lysed in RIPA buffer (50 mM Tris-HCl (pH 7.5), 1% Igepal, 0.5% sodium deoxycholate, 150 mM NaCl, and 0.5% sodium dodecyl sulfate). Phosphorylation of ERK1 / 2 in compound-treated lysates was assayed using the Phospho-ERK1 / 2 Whole Cell Lysate Kit (Meso Scale Discovery K151DWD) according to the manufacturer's protocol. Assay plates were read on a Meso Scale Discovery Sector Imager 6000 and data was analyzed using a 4-parameter logistic model to determine IC 50 The value was calculated.

[0322] GDP-KRAS G12D RF-MS Assay: The MS assay was performed in the following buffer: 25 mM HEPES (pH 7.4), 20 mM MgCl, 100 μM GDP freshly added to the assay buffer. 2 , 10 mM NaCl, 3% DMSO, 0.0005% CHAPS. The final concentration of KRAS G12D was 2 μM. The incubation time of the assay was 20 h and the inhibitor concentration was 100 μM. For the mass spectrometry assay, Greiner polypropylene clear V-bottom plates were used. Typically, the reaction was initiated by the addition of KRAS G12D to the wells containing the inhibitor, followed by Rapidfire MS analysis. The relative binding values ​​(%) at various incubation times were used to estimate the kobs value at 100 μM inhibitor.

[0323] device RapidFire sample liquid handling device: The RapidFire liquid handling device used was a RF 360 model. Solvents were delivered by an Agilent 1260 Infinity pump. The instrument control software was Agilent RapidFire High Throughput MS System Software (v4.0.12333.14274). A single C4 Type A (Agilent technologies, G9203) SPE-cartridge was used for these experiments. A volume of 10 μL (sample loop determined) was removed from each well of the 384-well plate by using an aspiration time of 300 ms. The sample load / wash time was 6000 milliseconds (ms) (H) at a flow rate of 1.25 mL / min. 2 2H2O, 0.1% v / v formic acid, 0.05% v / v trifluoroacetic acid); elution time was 7000 ms (acetonitrile, 0.1% v / v formic acid, 0.05% v / v trifluoroacetic acid); re-equilibration time was 700 ms (H2O, 0.1% v / v formic acid, 0.05% v / v trifluoroacetic acid) at a flow rate of 1.25 mL / min. 2 0, 0.1% v / v formic acid, 0.05% v / v trifluoroacetic acid).

[0324] Mass spectrometry: Mass spectrometry data were collected on an Agilent 6530 quadrupole time-of-flight (ToF) MS system operated with a dual Agilent Jet Spray (AJS) ion source in positive ionization mode. Instrument parameters were: gas temperature 350 °C; drying gas 5 L / min; nebulizer 60 psi; sheath gas 350 °C; sheath gas flow rate 11 L / min; capillary 3.5 kV (0.037 μA); nozzle 1.75 kV; fragmentator 200 V; skimmer 75 V; octopole RF 800 V peak-to-peak. Data were collected at a rate of 5 spectra / sec. The mass scale was calibrated using the Agilent positive ion tune mix over the m / z range of 300-3200. The instrument control software was Agilent MassHunter Workstation Data Acquisition (vB.05.01).

[0325] Data processing and informatics for mass spectrometry data: All data processing was performed using a combination of Agilent MassHunter Qualitative Analysis (B.05.00), Agilent RapidFire control software (v4.0.12333.14070) and Agilent DA Reprocessor Offline Utilities B.05.00, Build 162, on an HP Z620, 64-bit, 12-processor, Windows 7 PC. SPE-MS data collected from the RapidFire-MS system are typically single SPE-MS data files composed of multiple injection events. For efficient charge state extraction, SPE-peak integration and subsequent MS-spectral deconvolution, single SPE-MS data files are split into single injection events. This was typically performed offline and accomplished through the RapidFire user interface software using Agilent RapidFire control software (v4.0.12333.14070) using the Convert MS data option. Once the individual injection event MS-data files were generated, they were all batch processed using DA Reprocessor Offline Utilities (B.05.00, Build 162, Agilent). For batch processing, a method file was created using Agilent MassHunter Qualitative Analysis B.05.00. The details of this method file are contained within the BioConfirm Workflow and are as follows: Integrate (MS), select integrator: generally, smooth option was selected; detector point sampling, 1; start threshold, 5.0; stop threshold, 0.0; filtering, 5 points; peak location, top. Extract (MS): extract peak spectrum within m / z 800-1400 (MS).Deconvolute (MS) maximum entropy: mass range 19,000-21,000 Da (daltons); mass step 1.0 Da; S / N threshold 20.0; usage limit m / z range 800-1400; added protons; automatic isotope peak width; minimum successive charge state 5.

[0326] Once the individual injection event MS-files have been processed using the method files described above, a single report file is generated for each injection event MS file in the form of an Excel spreadsheet. The relevant protein molecular weights and intensity values ​​contained within specific cells in the spreadsheet are extracted from the Excel report / result files using a custom routine document in Pipeline Pilot (BIOVIA) and used to calculate percent control (POC) and percent binding (% bound) values ​​in the form of a concatenated output file (main text, tab separated in format; Figure S4a). POC=(uncorrected protein peak intensity / control protein peak intensity) x 100 and % bound=(predicted protein addition peak intensity / control protein peak intensity) x 100. Relative % bound=% bound / (POC+% bound) x 100 is a normalized measure of covalent binding that accounts for sample variability or compound effects on ionization or detection efficiency. These values ​​are then input into Amgen's data analysis suite (Genedata) for detailed data analysis and then finally loaded into Amgen's central database. Specific input files for the Pipeline Pilot script are an Excel report / result file directory; a compound molecular weight file (Amgen registered compound number and expected covalent modifications) and an injection sequence file (RFTIME file; not shown) that are generated during the RapidFire-MS acquisition and contain the injection sequence, injection location and plate barcode (matching the 384-well plate map dispensed by the Amgen Sample Bank). Additional information that is manually input into the Pipeline Pilot routine via the user interface is the molecular weight of the protein (average molecular weight (Da)) and the molecular weight variation, which is typically 0.01% (± 2 Da for a 19.5 kDa protein).

[0327] [Table 3]

[0328] [Table 4]

[0329] [Table 5]

[0330] The present invention is described in connection with the preferred embodiment. However, it should be understood that the present invention is not limited to the embodiment disclosed herein. It is understood that various modifications may be made by those skilled in the art once the description of the embodiment of the present invention is described herein. Such modifications are encompassed within the scope of the following claims.

Claims

1. Formula (I): 【Chemistry 1】 (In the formula, R １ is H, halo, or -CH3; R ２ is H, halo, or -CH3; R 3 is 【Chemistry 2】 【change】 is selected from R ８ is H, OH, or NH2) or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of said stereoisomer, or a pharma- ceutically acceptable salt of said atropisomer.

2. The structure: 【Chemistry 3】 2. The compound of claim 1 having the formula:

3. R １ The compound of claim 1 , wherein

4. R ２ The compound of claim 1 , wherein is Cl.

5. R ８ The compound of claim 1 , wherein

6. R ８ The compound of claim 1 , wherein is OH.

7. R ８ The compound of claim 1, wherein is NH2.

8. the below described: 【Chemistry 4】 【Chemistry 5】 【Chemistry 6】 【Chemistry 7】 【Chemistry 8】 【Chemistry 9】 or a stereoisomer thereof, an atropisomer thereof, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt of said stereoisomer, or a pharma- ceutically acceptable salt of said atropisomer.

9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 8 and a pharma- ceutically acceptable excipient.

10. 10. The pharmaceutical composition of claim 9 for use in the treatment of cancer.

11. The pharmaceutical composition of claim 10, wherein the cancer is non-small cell lung cancer, small intestine cancer, appendix cancer, colorectal cancer, endometrial cancer, pancreatic cancer, skin cancer, gastric cancer, nasal cancer or bile duct cancer.

12. The pharmaceutical composition of claim 11, wherein the cancer is non-small cell lung cancer.

13. The pharmaceutical composition of claim 11 , wherein the cancer is colorectal cancer.

14. The pharmaceutical composition of claim 11 , wherein the cancer is pancreatic cancer.

15. The pharmaceutical composition according to any one of claims 10 to 14, wherein the cancer is mediated by a KRAS G12C mutation.

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