Pyrimidopyridone and pteridinone derivatives as GCN2 kinase inhibitors, compositions and uses thereof

Abstract

The present application relates to pyrimidopyridone and pteridinone compounds of formula I, processes for their preparation and compositions comprising them. More particularly, the present application relates to compounds of formula I having activity as inhibitors of GCN2 kinase and their use in the treatment of diseases, disorders or conditions treatable by inhibition of GCN2 kinase, such as cancer and neuronal diseases. (I).

Description

[0001] This application claims the priority benefit of co-pending U.S. Provisional Patent Application 63 / 535,418, filed August 30, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to pyrimidopyridone and pteridinone compounds having activity as general regulators of GCN2 kinase inhibitors, methods for their preparation, compositions comprising them, and their use, for example, in treatment. More specifically, this application relates to compounds that can be used to treat diseases, disorders, or conditions (e.g., cancer and neuronal diseases) that can be treated by inhibiting GCN2 kinase. Background Technology

[0003] Eukaryotic initiation factor 2α (eIF2α) kinase generally regulates repressor protein 2 (GCN2), which drives cellular adaptation to amino acid restriction by activating and inducing an integrated stress response (ISR) of activated transcription factor 4 (ATF4). GCN2 kinase-mediated cellular adaptation to amino acid restriction occurs through translational control of gene expression, primarily executed via eIF2α phosphorylation. Using quantitative phosphorylated proteomics, Dokladal et al. recently demonstrated that GCN2 targets helper, physiologically relevant effectors, including eIF2β and Gcn20, to fine-tune translational control in response to amino acid starvation. Molecular Cell 2021; 81 (9), P1879-1889.e6). In addition to phosphorylating the eIF2-α subunit, GCN2 also phosphorylates the β-subunit of the trimer eIF2 G protein complex to promote its association with eIF5, which in turn helps to inhibit translation initiation.

[0004] Under unique stress conditions, cellular ISR is activated by four eukaryotic initiation factor 2α (eIF2α) kinases: GCN2, protein kinase-like endoplasmic reticulum kinase (PERK), double-stranded RNA-dependent kinase (PKR), and heme regulation inhibitor (HRI). Nat Rev Mol Cell Biol [2016, 17:213-226]. These four eIF2α kinases typically phosphorylate eIF2α at S51, thereby reducing overall protein synthesis. However, specific mRNAs with upstream open reading frames, such as activating transcription factor 4 (ATF4), are selectively translated by delayed translation restart via eIF2α phosphorylation. ATF4 is a key transcription factor for stress adaptation and subsequently drives gene transcription involved in processes such as protein folding, amino acid metabolism, and autophagy. Nat Rev Mol Cell Biol 2019, 20:436–450.

[0005] Within tumors, cancer cells frequently experience amino acid deprivation, partly due to abnormal proliferation increasing the demand for amino acids to produce proteins, lipids, and nucleic acids, and partly due to insufficient angiogenesis and disorder leading to amino acid supply shortages. Therefore, GCN2 may be important for cancer cell survival and tumor development. Furthermore, knockout of GCN2 or ATF4 has been shown to reduce tumor growth in vivo. EMBO. J. [2010, 29:2082–2096]. Furthermore, it has been shown that the GCN2 arm of the ISR protects cancer cells from intrinsic stress induced by the c-Myc oncogene. Nat Cell Biol 2019, 21:1413–1424; Nat Cell Biol 2019, 21:889–899). GCN2 may also be involved in resistance to cancer chemotherapy because in cancer cells expressing low levels of asparaginase synthase (ASNS), inhibition of GCN2 induces a sensitization effect against the antitumor agent L-asparaginase (L-ASNase). Proc Natl Acad Sci USA 2018, 115: E7776–E7785]. ASNS catalyzes the biosynthesis of asparagine (Asn) from aspartic acid and is highly responsive to cellular stress, particularly to intracellular amino acid depletion. ATF4 induces ASNS, [ J Biol Chem. [2017;292(49):19952-19958], which in turn maintains Asn levels and inhibits apoptosis, while the depletion of intracellular Asn induces apoptosis. Therefore, ASNS plays a role in tumor cell accumulation and progression by maintaining cell viability. Elevated ASNS protein expression is also associated with resistance to asparaginase therapy [ J Biol Chem. [2017;292(49):19952-19958]. Therefore, when used in combination with L-ASNase and GCN2 inhibition, tumors with high ASNS expression become sensitive to ASNS activity inhibition. This combination is a feasible strategy to control the growth, proliferation, and migration of cancer cells, eliminate them, or enhance their sensitivity to existing chemotherapeutic agents or radiotherapy. The GCN2-mediated ISR pathway has been proposed as a promising target for cancer therapy. Therefore, disrupting this pro-cancer stress-induced pathway by inhibiting GCN2 is an attractive therapeutic strategy.

[0006] Another important therapeutic area involving ISR activation is neuronal diseases or neuropathy. Science[2021, 373, 1161–1166]. Dominant mutations in universally expressed transfer RNA (tRNA) synthase genes lead to axonal peripheral neuropathy, constituting at least six types of Charcot-Marie-Tus disease (CMT). Genetic evidence in mouse and fruit fly models suggests the existence of gain-of-function mechanisms. It has been shown that mutant tRNA synthase activates the integrated stress response (ISR) via the sensor kinase GCN2. Chronic activation of ISR leads to pathophysiology, while gene deletion or pharmacological inhibition of GCN2 alleviates peripheral neuropathy. Activation of GCN2 suggests that the aberrant activity of mutant tRNA synthase remains translation-related, and that inhibition of GCN2 or ISR may represent a therapeutic strategy for CMT. Science 2021, 373, 1156–1161.

[0007] Recently, a small molecule inhibitor of GCN2 kinase (WO2021165346, Black Belt TXLTD) has been described.

[0008] There remains a need for effective GCN2 kinase inhibitors for the treatment of conditions such as cancer and peripheral neuropathy. Furthermore, there is a need for GCN2 kinase inhibitors that are selective compared to other kinases.

[0009] Overview The applicant has developed a novel, general regulatory inhibitor of repressor protein 2 (GCN2) kinase.

[0010] Therefore, the present invention includes compounds of formula I, or pharmaceutically acceptable salts, solvates, and / or prodrugs thereof: (I) in R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being surrounded by one or two R groups. 8 replace; X 1 Selected from N and CR 9 ; R 2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 2 Selected from N and CR 10 ; R 3 R 4 and R 5 Independently selected from H, halogens, CN, C1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 3 Selected from N and CR 11 ; R 6 and R 7 Independently selected from H, halogens, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, OC 1-6 Alkyl and OC 1-6 Halogenated alkyl groups; Each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 C(O)OR 12 =O, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, wherein all alkyl, alkenyl, ynyl, cycloalkyl, and heterocyclic alkyl groups are optionally selected from one or more halogens, OR 14 NR 14 R 15 and C 1-6 Alkyl substituents; R 9 R 10 and R 11 Independently selected from H, halogens, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; R 12 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being selected by one or two from halogen, OH, OC 1-4 Alkyl and OC 1-4 Substituents of fluoroalkyl groups; and R 13 R 14 and R 15 Independently selected from H and C 1-6 Alkyl and C 1-6 Halogenated alkyl groups.

[0011] This application also includes pharmaceutical compositions comprising one or more compounds of this application and a pharmaceutically acceptable carrier.

[0012] This application further includes a method for inhibiting generalized regulatory repressor protein 2 (GCN2) in cells in a biological sample or in a patient, comprising administering an effective amount of one or more compounds of this application to the cells.

[0013] This application also includes methods for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more compounds of this application to an individual in need.

[0014] This application also includes a method for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more compounds of this application to an individual in need in combination with another known agent that can be used to treat said diseases, disorders, or conditions that can be treated by inhibiting GCN2.

[0015] In some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer, and the one or more compounds of this application are combined with one or more additional cancer treatments (e.g., radiotherapy, chemotherapy (e.g., cisplatin), targeted therapies such as antibody therapy (including anti-PD1 and / or anti-PD-L1 antibodies) and small molecule therapies such as tyrosine kinase inhibitor therapy, glutaminase inhibitors (e.g., glutaminase-1 (GLS1) inhibitors) and asparagine synthase (ASNS) inhibitors, immunotherapy, hormone therapy, and anti-angiogenic therapy).

[0016] In some implementations, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer and / or peripheral neuropathy, including Charcot-Marie-Tuss (CMT) peripheral neuropathy.

[0017] This application also includes a method for improving the efficacy of one or more cancer treatments for treating cancer, comprising administering an effective amount of one or more compounds of this application in combination with an effective amount of the one or more cancer treatments.

[0018] Other features and advantages of this application will become apparent from the following detailed description. However, it should be understood that although embodiments of this application have been pointed out, the detailed description and specific examples are given by way of illustration only, and the scope of the claims should not be limited to these embodiments, but should be given the broadest interpretation consistent with the overall description. Invention Details I. Definition Unless otherwise specified, the definitions and embodiments described in this and other sections are intended to apply to all embodiments and aspects of this application to which they are appropriate, as will be understood by those skilled in the art.

[0020] The specification, including the claims, abstract, and drawings, discloses all features and all steps in any disclosed method or process, which may be combined in any combination except where at least some of such features and / or steps are mutually exclusive combinations. Unless otherwise expressly stated, each feature disclosed in the specification, including the claims, abstract, and drawings, may be replaced by an alternative feature for the same, equivalent, or similar purpose.

[0021] As used herein, the terms "compound of the application" or "compound of the present application" refer to compounds of formula I, including pharmaceutically acceptable salts, solvates, and / or prodrugs thereof.

[0022] As used herein, the terms "composition of the application" or "composition of the present application" mean a composition comprising one or more compounds of the present application and at least one additional ingredient.

[0023] As used herein, the term “and / or” means that the listed items are present or used individually or in combination. In practice, the term means that “at least one” or “one or more” of the listed items are used or present. The term “and / or” with respect to pharmaceutically acceptable salts and / or solvates means that the compounds of this application are present in combination as individual salts and hydrates, and solvates of salts of, for example, the compounds of this application.

[0024] As used in this application, the singular forms “a,” “an,” and “the” include plural references unless the context explicitly states otherwise. For example, an embodiment including “compound” should be understood to present certain aspects as one compound or two or more additional compounds.

[0025] In embodiments that include an “additional” or “second” component (such as an additional or second compound), as used herein, the second component is chemically different from the other components or the first component. A “third” component is different from the other, first and second components, and similarly different from the further enumerated or “additional” components.

[0026] As used herein, the terms “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “include” and “includes”), or “containing” (and any form of containing, such as “contain” and “contains”) are inclusive or open-ended and do not exclude additional unlisted elements or process / method steps.

[0027] As used herein, the term “composed of” and its derivatives are intended to be closed terms that specify the presence of the stated feature, element, component, group, integer and / or step, and exclude the presence of other unstated features, elements, components, groups, integers and / or steps.

[0028] As used herein, the term “substantially composed of” is intended to specify the presence of the stated features, elements, components, groups, integers and / or steps, as well as one or more essential and novel properties that do not substantially affect these features, elements, components, groups, integers and / or steps.

[0029] As used herein, degree terms such as “basically,” “about,” and “approximately” indicate a reasonable amount of deviation from the modifying term such that the final result is not significantly altered. These degree terms should be interpreted to include at least ±5% deviation from the modifying term if the deviation does not negate the meaning of the word it modifies.

[0030] As used herein, the term “suitable” means that the selection of a particular compound or conditions will depend on the specific synthetic operation to be performed, the characteristics of the molecule to be transformed, and / or the specific use of the compound, but such selection will be entirely within the skill level of a person trained in the art. All process / method steps described herein are carried out under conditions sufficient to provide the product shown. Those skilled in the art will understand that all reaction conditions, including, for example, reaction solvents, reaction times, reaction temperatures, reaction pressures, reactant ratios, and whether the reaction should be carried out under an anhydrous or inert atmosphere, can be modified to optimize the yield of the desired product, and to do so within their skill level.

[0031] This application references many chemical terms and abbreviations used by those skilled in the art. Nevertheless, for clarity and consistency, definitions of selected terms are provided.

[0032] As used herein, the terms “protecting group” or “PG” refer to a chemical motif that protects or masks the reactive parts of a molecule to prevent side reactions in those reactive parts, while simultaneously manipulating or causing different parts of the molecule to react. After the manipulation or reaction is complete, the protecting group is removed without degrading or breaking down the remaining parts of the molecule. Those skilled in the art can make the selection of a suitable protecting group. Many conventional protecting groups are known in the art, for example, as described in “Protective Groups in Organic Chemistry” McOmie, JFW ed., Plenum Press, 1973; Greene, TW and Wuts, PGM, “Protective Groups in Organic Synthesis”, John Wiley & Sons, 3rd ed., 1999; and Kocienski, P. Protecting Groups, 3rd ed., 2003, Georg Thieme Verlag (The Americas).

[0033] As used herein, the term “cell” refers to a single cell or multiple cells, including cells in a cell culture or an individual.

[0034] As used herein, the term "individual" includes all members of the animal kingdom, including mammals. Therefore, the methods and uses of this application are applicable to both human treatment and veterinary applications.

[0035] The term "pharmaceutically acceptable" means that it is compatible with an individual's treatment.

[0036] The term “pharmaceutically acceptable carrier” means a non-toxic solvent, dispersant, excipient, adjuvant or other material that mixes with an active ingredient (e.g., one or more compounds of this application) to allow the formation of a pharmaceutical composition (i.e., a dosage form that can be administered to an individual).

[0037] The term "pharmaceutically acceptable salt" means an acid or base addition salt that is suitable for or compatible with the individual's treatment.

[0038] An acid addition salt suitable for or compatible with the individual's treatment is any non-toxic organic or inorganic acid addition salt of any basic compound.

[0039] Suitable or compatible base addition salts for individual treatment are any non-toxic organic or inorganic base addition salts of any acidic compound.

[0040] As used herein, the term "prodrug" means a compound that is converted into an active drug upon administration, or a salt and / or solvate of a compound.

[0041] As used herein, the term "solvent" refers to a compound, or a salt or prodrug of a compound, wherein molecules of a suitable solvent are incorporated into the crystal lattice.

[0042] As used herein, the term "inert organic solvent" refers to a solvent that is generally considered not to react with functional groups present in compounds to be combined in any given reaction, such that it does not interfere with or inhibit the desired synthetic transformation. Organic solvents are typically nonpolar and dissolve compounds that are insoluble in aqueous solutions.

[0043] As used herein, the term "alkyl," whether used alone or as part of another group, refers to a straight-chain or branched saturated alkyl group. The possible number of carbon atoms in the alkyl group referred to is indicated by the prefix "C". n1-n2 "Indication. For example, the term C." 1-10 Alkyl refers to an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0044] As used herein, the term "halo" or "halogen," whether used alone or as part of another group, refers to a halogen atom and includes fluorine, chlorine, bromine, and iodine.

[0045] As used herein, the term "halogenated alkyl" refers to an alkyl group as defined above, in which one or more available hydrogen atoms are replaced by halogen atoms. Thus, for example, "C 1-6 "Halogenated alkyl" (or "C1-C6 halogenated alkyl") refers to a C1 to C6 straight-chain or branched alkyl group having one or more halogen substituents as defined above.

[0046] As used herein, the term "fluoroalkyl" refers to a haloalkyl group as defined above, wherein the halogen atom is fluorine.

[0047] As used herein, the term "chloroalkyl" refers to a haloalkyl group as defined above, wherein the halogen atom is chlorine.

[0048] The term "available" in phrases such as "available hydrogen atoms" or "available atoms" means an atom that is known to those skilled in the art to be substituted by another atom or group.

[0049] As used herein, the term "alkenyl," whether used alone or as part of another group, refers to a straight-chain or branched unsaturated alkyl group containing at least one double bond. The possible number of carbon atoms in the referred alkylene group is indicated by the prefix "C". n1-n2 "Indication. For example, the term C." 2-6 Alkenyl refers to an alkenyl group having 2, 3, 4, 5 or 6 carbon atoms and at least one double bond.

[0050] As used herein, the term "alkynyl," whether used alone or as part of another group, refers to a straight-chain or branched unsaturated alkynyl group containing at least one triple bond. The possible number of carbon atoms in the alkyl group referred to is indicated by the prefix "C". n1-n2 "Indication. For example, the term C." 2-6 The term "alkynyl" refers to an alkynyl group having 2, 3, 4, 5, or 6 carbon atoms.

[0051] As used herein, the term "cycloalkyl," whether used alone or as part of another group, refers to a saturated carbocyclic group containing 3 to 20 carbon atoms and one or more rings. The possible number of carbon atoms in the cycloalkyl group mentioned is indicated by the numerical prefix "C". n1-n2 "Indication. For example, the term C." 3-10 Cycloalkyl refers to cycloalkyl groups having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0052] As used herein, the term "heterocyclic alkyl," whether used alone or as part of another group, refers to a cyclic group containing at least one non-aromatic ring of 3 to 10 atoms, wherein one or more atoms are selected from O, S, S(O), SO2, N, NH, and N(C). 1-6 A heterocyclic alkyl group is a heterocyclic alkyl group with the remaining atoms being carbon atoms. Heterocyclic alkyl groups can be saturated or unsaturated (i.e., containing one or more double bonds). When a heterocyclic alkyl group contains the prefix C... n1-n2 In this case, the prefix indicates the number of carbon atoms in the corresponding carbocyclic group, wherein one or more (suitably 1 to 5) ring atoms are replaced by heteroatoms as defined above. The heterocyclic alkyl group is optionally benzofused.

[0053] All cyclic groups, including aryl, heteroaryl, heterocyclic, and cycloalkyl groups, contain one (i.e., monocyclic) or more than one ring (i.e., polycyclic). When a cyclic group contains more than one ring, the rings may be fused, bridged, or spirofused.

[0054] As used herein, the term "benzofused" refers to a polycyclic group in which the benzene ring is fused with another ring.

[0055] The term "fusion" between the first and second rings means that the first and second rings share two adjacent atoms between them.

[0056] The term "bridge" between the first and second rings means that the first and second rings share two non-adjacent atoms.

[0057] The term "spiro fusion" between the first and second rings means that the first and second rings share an atom between them.

[0058] As used herein, the term “optionally substituted” means that the group referred to is either unsubstituted or substituted.

[0059] As used herein, the term “substituted” refers to a substituent in which the atom in question contains at least one non-hydrogen atom.

[0060] When a group is substituted by one or more substituents, it should be understood that the choice of these substituents is independent of each other. That is, the one or more substituents can be the same or different.

[0061] When drawn as a symbol perpendicular to the key. Indicates the covalent bonding point of a chemical group.

[0062] As used in this article, the term "LCMS" refers to liquid chromatography-mass spectrometry.

[0063] As used in this article, the term "NMR" refers to nuclear magnetic resonance.

[0064] As used in this article, the term "aq." refers to water.

[0065] As used in this article, for example, the term "N" in "4N" refers to the symbol for the equivalent concentration unit "eq / L".

[0066] As used herein, for example in 4M, the term “M” refers to the molar concentration unit symbol that expresses “moles / L”.

[0067] As used in this article, the term "DIPEA" refers to N,N-diisopropylethylamine.

[0068] As used in this article, the term "DMF" refers to dimethylformamide.

[0069] As used in this article, the term "THF" refers to tetrahydrofuran.

[0070] As used in this article, the term "DMSO" refers to dimethyl sulfoxide.

[0071] As used in this article, the term "EtOAc" refers to ethyl acetate.

[0072] As used in this article, the term "MeOH" refers to methanol.

[0073] As used in this article, the term "EtOH" refers to ethanol.

[0074] As used in this article, the terms “MeCN” or “ACN” refer to acetonitrile.

[0075] As used in this article, the term "HCl" refers to hydrochloric acid.

[0076] As used in this article, the term "TFA" refers to trifluoroacetic acid.

[0077] As used in this article, the term "Hex" refers to hexane.

[0078] As used herein, the term "dppf" refers to 1,1'-bis(diphenylphosphine)ferrocene.

[0079] As used in this article, the term "RT" or "rt" refers to room temperature.

[0080] As used in this article, the term "HPLC" refers to high-performance liquid chromatography.

[0081] As used in this article, the terms “TEA” or “Et3N” refer to triethylamine.

[0082] As used in this article, the term "EDTA" refers to ethylenediaminetetraacetic acid.

[0083] As used in this article, the term "ATP" refers to adenosine triphosphate.

[0084] As used in this article, the term "FBS" refers to fetal bovine serum.

[0085] As used in this article, the term "MEM" refers to the minimum basal culture medium.

[0086] As used herein and as is known in the art, the terms “treating” or “treatment” mean a method of achieving a beneficial or desired outcome (including clinical outcomes). Beneficial or desired clinical outcomes may include, but are not limited to, alleviating or improving one or more symptoms or conditions, reducing the severity of a disease, disorder, or condition, stabilizing (i.e., not worsening) the state of a disease, disorder, or condition, preventing the spread of a disease, disorder, or condition, delaying or slowing the progression of a disease, disorder, or condition, improving or mitigating the state of a disease, disorder, or condition, reducing the recurrence of a disease, disorder, or condition, and alleviating (whether partially or completely), whether detectable or undetectable. “Treating” and “treatment” may also mean prolonged survival compared to expected survival in the absence of treatment. As used herein, “treating” and “treatment” also include preventative treatment.

[0087] "Palliance" of a disease, disorder, or symptom means a reduction in the severity and / or undesirable clinical manifestations of the disease, disorder, or symptom compared to an untreated disease, disorder, or symptom.

[0088] As used herein, the terms “prevention” or “control” or their synonyms refer to reducing an individual’s risk or likelihood of developing a disease, disorder, or condition that can be treated by inhibiting GCN2, or exhibiting symptoms associated with a disease, disorder, or condition that can be treated by inhibiting GCN2.

[0089] As used herein, the terms "effective amount" or "therapeutic effective amount" mean the amount of one or more compounds of this application that effectively achieves the desired results within the necessary dosage and time period.

[0090] The term "disease, disorder, or condition that can be treated by inhibiting GCN2" means that the disease, disorder, or condition to be treated is influenced or regulated by GCN2 activity, particularly increased GCN2 activity, and / or has some direct or indirect biological basis, including GCN2 activity, particularly increased GCN2 activity. When the GCN2 activity associated with the disease, disorder, or condition is inhibited by one or more compounds or compositions of this application, these diseases produce a beneficial response.

[0091] As used herein, the term "inhibition of GCN2" refers to the inhibition, blocking, and / or disruption of the kinase activity or function of GCN2 in cells. Inhibition, blocking, and / or disruption induce a therapeutic effect in cells.

[0092] The term "inhibition, blocking and / or destruction" refers to any detectable inhibition, blocking and / or destruction in the presence of a compound, compared to conditions identical in all respects except the absence of the compound.

[0093] As used herein, the term “GCN2” refers to General Control Nonderepressible 2, or any functional mutant or similar form thereof.

[0094] As used in this article, the term "low asparagine synthase (ASNS) expression" means any detectable reduction or decrease in the level of asparagine synthase (ASNS) in cancer cells, all other things being equal except in healthy cells.

[0095] As used in this article, the expression “asparagine synthase (ASNS) overexpression or dysregulation” means any detectable increase in the level of asparagine synthase (ASNS) in cancer cells, all other things being equal, except in healthy cells.

[0096] As used in this article, the term "low glutaminase expression" means any detectable reduction or decrease in the level of glutaminase (e.g., GLS1) in cancer cells, other than in healthy cells, under otherwise identical conditions.

[0097] As used in this article, the expression “glutaminase overexpression or dysregulation” means any detectable increase in glutaminase (e.g., GLS1) levels in cancer cells, other than in healthy cells, under otherwise identical conditions.

[0098] The term “GLS1” as used in this article refers to “renal type” glutaminase or any functional mutant or similar form thereof.

[0099] As used herein, the term “application” means the application of a therapeutically effective amount of the compound of this application, or one or more compounds, or a composition thereof, to a cell or individual.

[0100] As used herein, the term "neoplastic condition" refers to a disease, disorder, or symptom characterized by cells with autonomous growth or replication capabilities, such as an abnormal state or symptom characterized by proliferating cell growth. As used herein, the term "growth" refers to a mass of tissue resulting from the abnormal growth and / or division of cells in an individual with a neoplastic condition. Growths can be benign (e.g., uterine fibroids and melanocytic nevi), potentially malignant (e.g., carcinoma in situ), or malignant (e.g., cancer).

[0101] As used in this article, the term "fibrosis" refers to a disease, disorder, or condition characterized by thickening and scarring of connective tissue, which is usually a result of injury.

[0102] II. Compounds of this application The pyrimidopyridone and pteridone compounds of this application were prepared and found to inhibit the kinase generally regulating repressor protein 2 (GCN2).

[0103] Therefore, this application includes compounds of formula I, or pharmaceutically acceptable salts, solvates, and / or prodrugs thereof: (I) in R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being surrounded by one or two R groups. 8 replace; X 1 Selected from N and CR 9 ; R 2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 2 Selected from N and CR 10 ; R 3 R 4 and R 5 Independently selected from H, halogens, CN, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X3 Selected from N and CR 11 ; R 6 and R 7 Independently selected from H, halogens, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, OC 1-6 Alkyl and OC 1-6 Halogenated alkyl groups; Each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 C(O)OR 12 =O, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, wherein all alkyl, alkenyl, ynyl, cycloalkyl, and heterocyclic alkyl groups are optionally selected from one or more halogens, OR 14 NR 14 R 15 and C 1-6 Alkyl substituents; R 9 R 10 and R 11 Independently selected from H, halogens, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; R 12 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being selected by one or two from halogen, OH, OC 1-4 Alkyl and OC 1-4 Substituents of fluoroalkyl groups; and R 13 R 14 and R 15 Independently selected from H and C 1-6 Alkyl and C 1-6 Halogenated alkyl groups.

[0104] This application also includes compounds of formula I, or pharmaceutically acceptable salts, solvates, and / or prodrugs thereof: (I) in R1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being surrounded by one or two R groups. 8 replace; X 1 Selected from N and CR 9 ; R 2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 2 Selected from N and CR 10 ; R 3 R 4 and R 5 Independently selected from H, halogens, CN, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 3 Selected from N and CR 11 ; R 6 and R 7 Independently selected from H, halogens, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, OC 1-6 Alkyl and OC 1-6 Halogenated alkyl groups; Each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 Halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, wherein all alkyl, alkenyl, ynyl, cycloalkyl, and heterocyclic alkyl groups are optionally selected from one or more halogens, OR 14 NR 14 R 15 and C 1-6 Alkyl substituents; R 9 R 10 and R 11 Independently selected from H, halogens, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; R 12 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being selected by one or two from halogen, OH, OC 1-4 Alkyl and OC 1-4 Substituents of fluoroalkyl groups; and R 13 R 14 and R 15 Independently selected from H and C 1-6 Alkyl and C 1-6 Halogenated alkyl groups.

[0105] In some implementation schemes, R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being replaced by an R 8 Replacement. In some implementations, R 1 Selected from C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, each optionally separated by one or two R 8 replace.

[0106] In some implementation schemes, R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being replaced by an R 8 Replacement. In some implementations, R 1 Selected from C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, each optionally separated by one or two R 8 replace.

[0107] In some implementation schemes, R 1 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups. In some embodiments, R 1 Selected from H, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups. In some embodiments, R 1 Selected from H, C 1-4 Alkyl and C 1-4 Fluoroalkyl. In some embodiments, R 1Selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3, and CH(CH3)3. In some embodiments, R 1 Selected from H, CH3, CH2CH3 and CH(CH3)2.

[0108] In some implementation schemes, R 1 It can be arbitrarily selected by one or two Rs. 8 Replacement C 3-10 Cycloalkyl. In some embodiments, R 1 It is a single-ring C 3-10 cycloalkyl or bicyclic C 5-10 Cycloalkyl groups, each optionally bound by one or two R 8 Replacement. In some implementations, R 1 It can be arbitrarily selected by one or two Rs. 8 Substituted monocyclic C3-8 cycloalkyl groups. In some embodiments, R 1 The single-ring C in 3-8 The cycloalkyl group is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, each optionally separated by one or two R groups. 8 Replacement. In some implementations, R 1 Selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, each of which is randomly diffused by an R 8 Replacement. In some implementations, R 1 The compound is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, each of which is unsubstituted. In some embodiments, R 1 It is cyclopropyl. In some implementations, R 1 Selected from cyclobutyl, cyclopentyl, and cyclohexyl, each separated by one or two R groups. 8 Replacement. In some implementations, R 1 Is it by one or two Rs 8 Replaced cyclohexyl. In some implementations, R 1 Selected from cyclobutyl and cyclohexyl, each is converted by an R 8 Replacement. In some implementations, R 1 It was by an R 8 Substituted cyclobutyl. In some embodiments, R 1 It was by an R 8 Substituted cyclohexyl groups.

[0109] In some implementation schemes, R 1 It is a spiro-fused C 5-10 cycloalkyl or bridged C 5-10 Cycloalkyl groups, each optionally bound by one or two R 8Replacement. In some embodiments, the spiro-fused C 5-10 The cycloalkyl group is selected from spiro[3.3]heptane, spiro[4.4]nonane, spiro[5.4]decane, spiro[4.5]octane and spiro[5.2]octane, each optionally distilled by one or two R groups. 8 Replacement. In some embodiments, the spiro-fused C 6-10 Cycloalkyl is optionally enclosed by one or two R 8 Substituted spiro[3.3]heptane. In some embodiments, the spirofused C 5-10 cycloalkyl is .

[0110] In some implementation schemes, R 1 It can be arbitrarily selected by one or two Rs. 8 Replacement bridge C 5-10 Cycloalkyl. In some embodiments, the bridging C 5-10 The cycloalkyl group is selected from bicyclopentyl, bicycloheptyl, and bicyclooctyl, each optionally marked by one or two R... 8 Replacement. In some implementations, the bridge C 5-10 The cycloalkyl group is selected from bicyclopentyl, bicycloheptyl, and bicyclooctyl, each optionally dimethyl alkyl (R) 8 Replacement. In some implementations, the bridge C 5-10 cycloalkyl groups are selected from , and .

[0111] In some implementation schemes, R 1 It can be arbitrarily selected by one or two Rs. 8 Replacement C 3-10 Heterocyclic alkyl groups. In some embodiments, R 1 It is arbitrarily selected by an R 8 Replacement C 3-10 Heterocyclic alkyl groups. In some embodiments, R 1 It is arbitrarily selected by an R 8 Replacement C 3-6 Heterocyclic alkyl groups. In some embodiments, R 1Selected from azircyclopropane, oxacyclopropane, thiohexacyclopropane, azircyclobutane, oxacyclobutane, thiohexacyclobutane, diazircyclobutane, dioxacyclobutane, dithiohexacyclobutane, tetrahydrofuranyl, tetrahydrothiopheneyl, pyrrolidinyl, imidazoalkyl, pyrazolyl, isoxthiolidinyl, thiazoalkyl, isothiazolyl, dioxacyclopentyl, dithiohexacyclopentyl, piperidinyl, tetrahydropyranyl, diazinanyl (e.g., piperazinyl), morpholinyl, thiomorpholinyl, dioxyl, dithiadinyl, azircycloheptyl, oxacycloheptyl, and thiohexacycloheptyl, each optionally separated by one or two R 8 Replacement. In some implementations, R 1 Selected from thioheterobutyl, oxoheterobutyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl, azirropropyl, azirrobutyl, pyrrolidinyl, morpholinyl, piperazine, and piperidinyl, each optionally distilled by an R 8 Replacement. In some implementations, R 1 Selected from oxetane, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, piperazineyl, and piperidinyl, each optionally distilled by an R 8 Replacement. In some implementations, R 1 Selected from oxetane, tetrahydrofuranyl, and tetrahydropyranyl, each optionally converted by an R 8 Replacement. In some implementations, R 1 It is selected from oxetane, tetrahydrofuranyl and tetrahydropyranyl, each of which is unsubstituted.

[0112] In some implementation schemes, X 1 Selected from N and CH. In some implementations, X 1 For N. In some implementations, X 1 For CH.

[0113] In some implementation schemes, R 2 Selected from H, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups. In some embodiments, R 2 Selected from H, C 1-4 Alkyl and C 1-4 Fluoroalkyl. In some embodiments, R 2 Selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3, and CH(CH3)3. In some embodiments, R 2 Selected from H, CH3, CH2CH3, and CH(CH3)2. In some embodiments, R 2Selected from H, CH3, and CH(CH3)2. In some embodiments, R 2 It consists of H and CH3.

[0114] In some implementation schemes, X 2 Selected from N and CH. In some implementations, X 1 It is N. In some implementations, X 2 It is CH.

[0115] In some implementation schemes, R 3 R 4 and R 5 Not all are H. In some implementations, R 3 R 4 and R 5 Independently selected from H, halogens, CN, C 1-4 Alkyl and C 1-4 Halogenated alkyl groups. In some embodiments, R 3 R 4 and R 5 At least one of them is selected from halogen, CN and C. 1-4 Halogenated alkyl groups. In some embodiments, R 3 and R 5 At least one of them is selected from halogen, CN and C. 1-4 Halogenated alkyl groups. In some embodiments, R 3 R 4 and R 5 At least one of them is selected from halogen and CN. In some embodiments, R 3 and R 5 At least one of them is selected from halogen and CN. In some embodiments, R 3 R 4 and R 5 At least one of them is a halogen. In some embodiments, R 3 and R 5 At least one of them is a halogen. In some embodiments, R 3 R 4 and R 5 Independently selected from H, Cl, F, Br, CN, C 1-4 Alkyl and C 1-4 Fluoroalkyl. In some embodiments, R 3 R 4 and R 5 At least one of them is selected from F, Cl, Br, CN and C. 1-4 Fluoroalkyl. In some embodiments, R 3 and R 5 At least one of them is selected from F, Cl, Br, CN and C. 1-4Fluoroalkyl. In some embodiments, R 3 R 4 and R 5 It is independently selected from H, Cl, F, CN, CH3, and CF3. In some embodiments, R 3 R 4 and R 5 At least one of them is selected from Cl, F, CN, and CF3. In some embodiments, R 3 R 4 and R 5 At least one of them is selected from Cl, F, and CN. In some embodiments, R 3 and R 5 At least one of them is selected from Cl, F, and CN. In some embodiments, R 3 R 4 and R 5 Independently selected from H, F, and CN. In some implementations, R 3 R 4 and R 5 At least one of them is selected from F and CN. In some embodiments, R 3 and R 5 At least one of them is selected from F and CN. In some embodiments, R 3 and R 5 At least one of them is selected from F and CN, and R 4 It is H. In some implementations, R 3 It is CN, R 4 It is H and R 5 It is H. In some implementations, R 3 It is H, R 4 It is H and R 5 It is CN. In some implementations, R 3 and R 5 Independently selected from H, F, and CN, and R 4 It is H. In some implementations, R 3 and R 5 Independently selected from H and F, and R 4 It is H. In some implementations, R 3 and R 5 At least one of them is F. In some implementations, R 3 and R 5 At least one of them is F and R 4 It is H. In some implementations, R 3 and R 5 Both are F and R 4 It is H. In some implementations, R 3 It is F, R 4 It is H and R5 It is F. In some implementations, R 3 It is F, R 4 It is H and R 5 It is H. In some implementations, R 3 It is H, R 4 It is H and R 5 It is F.

[0116] In some implementation schemes, X 3 Selected from N and CH. In some implementations, X 3 It is CH.

[0117] In some implementation schemes, R 6 and R 7 Independently selected from H, Cl, F, Br, CN, C 1-4 Alkyl, C 1-4 Fluoroalkyl, OC 1-4 Alkyl and OC 1-4 Fluoroalkyl. In some embodiments, R 6 and R 7 Independently selected from H, Cl, F, CN, CH3, CHF2, CF3, CH2CH3, OCH3, OCHF2, and OCF3. In some embodiments, R 6 and R 7 It is independently selected from Cl, F, CH3, CF3, CHF2, CH2CH3, OCH3, OCHF2, and OCF3. In some embodiments, R 6 Selected from OCH3 and OCF3, and R 7 Selected from Cl, F, CH3, and CF3. In some embodiments, R 6 Selected from OCH3 and OCF3, and R 7 It is Cl. In some implementations, R 6 It is OCH3 and R 7 It is Cl.

[0118] In some implementation schemes, each R 8 Selected independently from OR 12 C(O)NR 12 R 13 C(O)OR 12 =O,NR 12 R 13 Halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-10 cycloalkyl and C 3-10Heterocyclic alkyl groups, wherein all alkyl, alkenyl, ynyl, cycloalkyl, and heterocyclic alkyl groups are optionally selected from one or more halogens, OR 14 NR 14 R 15 and C 1-4 Alkyl substituents. In some embodiments, each R... 8 Selected independently from OR 12 C(O)NR 12 R 13 NR 12 R 13 Halogen, C 1-4 Alkyl, C 1-4 Haloalkyl, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, wherein all alkyl, alkenyl, ynyl, cycloalkyl, and heterocyclic alkyl groups are optionally selected from one or more halogens, OR 14 NR 14 R 15 and C 1-4 Alkyl substituents. In some embodiments, each R... 8 Selected independently from OR 12 C(O)NR 12 R 13 C(O)OR 12 =O,NR 12 R 13 Cl, F, Br, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-6 cycloalkyl and C 3-6 Heterocyclic alkyl groups, wherein all alkyl, cycloalkyl, and heterocyclic alkyl groups are optionally separated by one or more elements selected from Cl, Br, F, and OR. 14 NR 14 R 15 and C 1-4 Alkyl substituents. In some embodiments, each R... 8 Selected independently from OR 12 C(O)NR 12 R 13 NR 12 R 13 Cl, F, Br, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-6 cycloalkyl and C 3-6 Heterocyclic alkyl groups, wherein all alkyl, cycloalkyl, and heterocyclic alkyl groups are optionally separated by one or more elements selected from Cl, Br, F, and OR. 14 NR 14R 15 and C 1-4 Alkyl substituents. In some embodiments, each R... 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 C(O)OR 12 =O, Cl, F, CH3, CHF2, CH3CH3, and CF3. In some implementations, each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 Cl, F, CH3, CHF2, CH3CH3, and CF3. In some implementations, each R 8 The components are independently selected from Cl, F, CH3, CHF2, CH3CH3, and CF3. In some embodiments, each R... 8 The components are independently selected from CH3, CHF2, CH3CH3, and CF3. In some embodiments, each R... 8 Selected independently from OR 12 NR 12 R 13 and C(O)NR 12 R 13 In some implementation schemes, each R 8 Independently selected from NR 12 R 13 and C(O)NR 12 R 13 In some implementation schemes, each R 8 Independently selected from =O and C(O)OR 12 In some implementation schemes, each R 8 Yes = O. In some implementations, each R 8 It is C(O)OR 12 In some implementations, when each R 8 Selected independently from OR 12 C(O)NR 12 R 13 C(O)OR 12 =O,NR 12 R 13 C 3-10 cycloalkyl and C 3-10 In heterocyclic alkyl groups, R1 is only separated by one R 8 replace.

[0119] In some implementation schemes, R 9R 10 and R 11 Independently selected from H, Cl, Br, F, C 1-4 Alkyl and C 1-4 Fluoroalkyl. In some embodiments, R 9 R 10 and R 11 Independently selected from H, Cl, F, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3, and CH(CH3)3. In some embodiments, R 9 R 10 and R 11 It is independently selected from H, Cl, F, CH3, and CF3. In some embodiments, R 9 R 10 and R 11 Independently selected from H and F. In some implementations, R 9 R 10 and R 11 It is H.

[0120] In some implementation schemes, R 12 Selected from H, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally substituted with one or two substituents selected from Cl, F, Br, OH, OCH3, and OCF3. In some embodiments, R 12 Selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3, and CH(CH3)3. In some embodiments, R 12 Selected from H, CH3, and CF3. In some implementations, R 12 Selected from H and CH3. In some implementations, R 12 It is CH3.

[0121] In some implementation schemes, R 13 Selected from H, C 1-4 Alkyl and C 1-4 Fluoroalkyl. In some embodiments, R 13 Selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3, and CH(CH3)3. In some embodiments, R 13 Selected from H, CH3, and CF3. In some implementations, R 13 Selected from H and CH3.

[0122] In some implementation schemes, R 14 and R 15 Independently selected from H and C 1-4 Alkyl and C 1-4 Fluoroalkyl. In some embodiments, R 14 and R 15 Independently selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3, and CH(CH3)3. In some embodiments, R 14 and R 15 It is independently selected from H, CH3, and CF3. In some implementations, R 14 and R 15 It is independently selected from H and CH3.

[0123] In some embodiments, the compound of formula I is selected from:

[0124] Or its pharmaceutically acceptable salts, solvates and / or prodrugs.

[0125] In some embodiments, the pharmaceutically acceptable salt is an acid addition salt or a base addition salt. The selection of a suitable salt can be made by those skilled in the art (see, for example, SM Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19).

[0126] Acid addition salts suitable for or compatible with the individual's treatment are any non-toxic organic or inorganic acid addition salts of any basic compound. Basic compounds forming acid addition salts include, for example, compounds containing an amino group. Exemplary inorganic acids forming suitable salts include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, as well as acidic metal salts such as sodium monohydrogen phosphate and potassium hydrogen sulfate. Exemplary organic acids forming suitable salts include mono-, di-, and tricarboxylic acids. Exemplary examples of such organic acids are, for example, acetic acid, formic acid, trifluoroacetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid, hydroxybenzoic acid, phenylacetic acid, cinnamic acid, mandelic acid, salicylic acid, 2-phenoxybenzoic acid, p-toluenesulfonic acid, and other sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, and 2-hydroxyethanesulfonic acid. In one embodiment, a monoacid or diacid salt is formed, and such salts exist in a hydrated, solvated, or substantially anhydrous form. Generally, acid addition salts are more soluble in water and a variety of hydrophilic organic solvents and typically exhibit a higher melting point compared to their free base form. The selection criteria for suitable salts are known to those skilled in the art. Other non-pharmaceuticalally acceptable salts (such as, but not limited to, oxalates) may be used, for example, to isolate the compounds of this application for laboratory use or subsequently convert them into pharmaceutically acceptable acid addition salts. In some embodiments, the acid addition salt is a hydrochloric acid or formic acid addition salt.

[0127] Suitable or therapeutically compatible base addition salts are any non-toxic organic or inorganic base addition salts of any acidic compound. Acidic compounds that form base addition salts include, for example, compounds containing a carboxylic acid group. Exemplary inorganic bases that form suitable salts include lithium hydroxide, sodium, potassium, calcium, magnesium, or barium, and ammonia. Exemplary organic bases that form suitable salts include aliphatic, alicyclic, or aromatic organic amines such as isopropylamine, methylamine, trimethylamine, methylpyridine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, meglumine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Exemplary organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. Suitable salts may be chosen to prevent the hydrolysis of ester functional groups (if any) elsewhere in the compound. Criteria for choosing suitable salts are known to those skilled in the art.

[0128] The solvates of the compounds in this application include those prepared, for example, with pharmaceutically acceptable solvents. Examples of such solvents include water (the resulting solvate is referred to as hydrate) and ethanol. Suitable solvents are physiologically tolerable at the administered dose.

[0129] The compounds of this application may also exist in different tautomer forms, and any tautomer forms formed by the compounds, as well as mixtures thereof, are intended to be included within the scope of this application.

[0130] The compounds of this application may also exist in different polymorphs, and it is anticipated that any polymorph or mixture thereof is included within the scope of this application.

[0131] The compounds of this application may be further radiolabeled, therefore all radiolabeled forms of the compounds of this application are included within the scope of this application. The compounds of this application also include those compounds incorporating one or more radioactive atoms in their structure.

[0132] III. The Composition of this Application The compounds of this application are suitably formulated into compositions in a conventional manner using one or more carriers. Therefore, this application also includes compositions comprising one or more compounds of this application and a carrier. The compounds of this application are suitably formulated into pharmaceutical compositions for administration to an individual in a biologically compatible form suitable for in vivo administration. Therefore, this application also includes pharmaceutical compositions comprising one or more compounds of this application and a pharmaceutically acceptable carrier. In embodiments of this application, the pharmaceutical compositions are used to treat any of the diseases, disorders, or conditions described herein.

[0133] Those skilled in the art will understand that the compounds of this application can be administered to an individual in a variety of forms, depending on the chosen route of administration. For example, the compounds of this application can be administered orally, by inhalation, parenteral administration, sublingual administration, nasal administration, rectal administration, vaginal administration, patch administration, pump administration, microvacuum pump administration, topical or transdermal administration, and correspondingly formulated pharmaceutical compositions. In some embodiments, administration is performed by pump administration in a periodic or continuous manner. Conventional procedures and ingredients for selecting and preparing suitable compositions are described, for example, in Remington's Pharmaceutical Sciences (2000–20th edition) and The United States Pharmacopeia: The National Formulary (USP 24 NF19), published in 1999.

[0134] Parenteral administration includes systemic delivery routes other than the gastrointestinal (GI) tract, including, for example, intravenous, intra-arterial, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary (e.g., by using an aerosol), intrathecal, rectal, and local (including by using patches or other transdermal delivery devices) administration modes. Parenteral administration can be performed by continuous infusion over a selected time period.

[0135] In some embodiments, the compounds of this application are administered orally, for example, with an inert diluent or with an assimilated edible carrier, or encapsulated in hard or soft-shell gelatin capsules, or compressed into tablets, or directly incorporated into dietary foods. In some embodiments, the compounds are incorporated into excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, capsules, pills, granules, tablets, chewing gum, powders, syrups, elixirs, rice paper capsules, aqueous solutions, and suspensions. For tablets, carriers used include lactose, corn starch, sodium citrate, and phosphates. Pharmaceutically acceptable excipients include binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potato starch or sodium carboxymethyl starch); or wetting agents (e.g., sodium lauryl sulfate). In embodiments, tablets are coated using methods known in the art. In the case of tablets, capsules, pouches, pills, or granules for oral administration, a pH-sensitive enteric coating, such as Eudragits™ designed to control the release of the active ingredient, is optionally used. Oral dosage forms also include modified release formulations (e.g., immediate release and timed release). Examples of modified release formulations include sustained release (SR), extended release (ER, XR, or XL), delayed release or timed release, controlled release (CR), or continuous release (CR or Contin) in the form of, for example, coated tablets, permeation delivery devices, coated capsules, microencapsulated microspheres, aggregated particles (e.g., as molecular sieve-type particles), or fine hollow permeable fiber bundles, or chopped hollow permeable fibers aggregated or contained in fiber pouches. Timed release compositions are formulated, for example, into liposomes, or those in which the active compound is protected, for example, by microencapsulation, multiple coatings, etc., with a differentially degradable coating. Liposome delivery systems include, for example, small monolayer vesicles, large monolayer vesicles, and multilayer vesicles. In some implementations, liposomes are formed from various phospholipids, such as cholesterol, stearamine, or phosphatidylcholine. For oral administration in capsule form, useful carriers or diluents include lactose and dried corn starch.

[0136] In some embodiments, the liquid formulation for oral administration is in the form of, for example, a solution, syrup, or suspension, or suitably presented as a dry product prepared with water or other suitable medium prior to use. When an aqueous suspension and / or emulsion is administered orally, the compound of this application is suitably suspended or dissolved in an oil phase in combination with an emulsifier and / or suspending agent. If desired, certain sweeteners and / or flavoring agents and / or coloring agents are added. Such a liquid formulation for oral administration is prepared by conventional means using pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methylcellulose, or hydrogenated edible fats); emulsifiers (e.g., lecithin or gum arabic); non-aqueous mediums (e.g., almond oil, oily esters, or ethanol); and preservatives (e.g., methylparaben or propylparaben or sorbic acid). Available diluents include lactose and high molecular weight polyethylene glycol.

[0137] The compounds of this application can also be freeze-dried, and the obtained freeze-dried products can be used, for example, to prepare injectable products.

[0138] In some embodiments, the compounds of this application are for parenteral administration. For example, solutions of the compounds of this application are prepared in water appropriately mixed with a surfactant such as hydroxypropyl cellulose. In some embodiments, dispersions are prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof with or without alcohol, as well as oils. Under normal storage and use conditions, these formulations contain preservatives to prevent microbial growth. Those skilled in the art know how to prepare suitable formulations. For parenteral administration, sterile solutions of the compounds of this application are typically prepared, and the pH of the solution is appropriately adjusted and buffered. For intravenous use, the total concentration of the solute should be controlled to make the formulation isotonic. For ocular application, ointments or drop-able liquids are delivered via ocular delivery systems known in the art, such as applicators or eye drops. In some embodiments, such compositions include mucus mimics such as hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose, or polyvinyl alcohol, preservatives such as sorbic acid, EDTA, or benzyl chromium chloride, and a conventional amount of diluent or carrier. For pulmonary application, the diluent or carrier will be selected to appropriately allow the formation of an aerosol.

[0139] In some embodiments, the compounds of this application are formulated for parenteral administration by injection (including using conventional catheter insertion techniques or infusion). For example, the injectable formulation is provided in unit dosage form, such as in ampoules or multi-dose containers with added preservatives. In some embodiments, the composition is in the form of a sterile suspension, solution, or emulsion, such as in an oily or aqueous medium, and contains formulations such as suspending agents, stabilizers, and / or dispersants. In all cases, the form must be sterile and must be fluid to achieve a degree of injectability. Alternatively, the compounds of this application may suitably be in sterile powder form to be reconstituted with a suitable medium (e.g., sterile pyrogen-free water) prior to use.

[0140] In some embodiments, the composition for nasal application is conveniently formulated as an aerosol, drops, gel, or powder.

[0141] For intranasal or inhalation administration, the compounds of this application are conveniently delivered in the form of a solution, dry powder, or suspension from a pump-operated spray container squeezed or pumped by a patient, or in the form of an aerosol spray from a pressurized container or nebulizer. Aerosol formulations typically comprise a solution or fine-particle suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent, and are generally presented in a sterile, sealed container in single or multiple doses, for example, in cartridge or refill form for use with a nebulizer. Alternatively, the sealed container may be an integrated dispensing device such as a single-dose nasal inhaler, or an aerosol dispenser equipped with a metering valve designed for post-use disposal. In dosage forms that include an aerosol dispenser, it may contain a propellant, such as a compressed gas like compressed air, or an organic propellant like chlorofluorocarbons (CFCs). Suitable propellants include, but are not limited to, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, heptafluoroalkane, carbon dioxide, or another suitable gas. In the case of pressurized aerosols, the dosage unit is suitably determined by providing a valve for delivering the measured amount. In some embodiments, the pressurized container or nebulizer contains a solution or suspension of the active compound. Capsules and cartridges (made of, for example, gelatin) for inhalers or blowpipes are formulated, for example, into a powder mixture containing the compound of this application and a suitable powder base (such as lactose or starch). Aerosol dosage forms can also be in the form of a pump-nebulizer.

[0142] Compositions suitable for sublingual or oral administration include tablets, lozenges, and confectionery lozenges, wherein the compounds of this application are formulated with a carrier such as sugar, gum arabic, astragalus gum, or gelatin and glycerin. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0143] The suppository form of the compounds of this application is for vaginal, urethral, ​​and rectal administration. Such suppositories are typically constructed from mixtures of substances that are solid at room temperature but molten at body temperature. Commonly used substances to form such media include, but are not limited to, cocoa butter (also known as cocoa grease), glycerin gelatin, other glycerides, hydrogenated vegetable oils, and mixtures of polyethylene glycol and its fatty acid esters of various molecular weights. For further discussion of suppository dosage forms, see, for example: Remington's Pharmaceutical Sciences , 16th edition, Mack Publishing, Easton, PA, 1980, pp. 1530-1533.

[0144] In some embodiments, the compounds of this application are coupled to a soluble polymer that serves as a targeted drug carrier. Such polymers include, for example, polyvinylpyrrolidone, pyran copolymers, polyhydroxypropyl methacrylamide-phenol, polyhydroxyethyl asparagine, or polyethylene oxide polylysine substituted with palmitoyl residues. Furthermore, in some embodiments, the compounds of this application are coupled to a class of biodegradable polymers that can be used to achieve controlled drug release, such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, polycaprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyran, polycyanoacrylates, and crosslinked or amphiphilic block copolymers of hydrogels.

[0145] In some embodiments, the compounds of this application may be conjugated to viral, non-viral, or other vectors. Viral vectors may include retroviruses, lentiviruses, adenoviruses, herpesviruses, poxviruses, alphaviruses, vaccinia viruses, or adeno-associated viruses. Non-viral vectors may include nanoparticles, cationic lipids, cationic polymers, metal nanoparticles, nanorods, liposomes, micelles, microvesicles, cell-penetrating peptides, or lipospheres. Nanoparticles may include silica, lipids, carbohydrates, or other pharmaceutically acceptable polymers.

[0146] The compounds of this application (including their pharmaceutically acceptable salts and / or solvates) are suitably used alone, but are generally administered in the form of pharmaceutical compositions in which one or more of the compounds of this application (active ingredients) are bound to a pharmaceutically acceptable carrier. Depending on the administration method, the pharmaceutical composition may comprise about 0.05% to about 99% by weight or about 0.10% to about 70% by weight of the active ingredient, and about 1% to about 99.95% by weight or about 30% to about 99.90% by weight of a pharmaceutically acceptable carrier, all weight percentages being based on the total composition.

[0147] IV. Methods and Uses of this Application It has been shown that the compounds of this application inhibit or block the general regulatory repressor protein 2 (GCN2) kinase, thereby weakening the transcriptional function of ATF4 target gene expression. Therefore, the compounds of this application can be used to inhibit GCN2.

[0148] Therefore, this application includes a method for inhibiting general regulatory repressor protein 2 (GCN2) in cells in a biological sample or in a patient, comprising administering an effective amount of one or more compounds of this application to the cells.

[0149] This application also includes the use of one or more of the compounds of this application for inhibiting GCN2 in cells, and the use of one or more of the compounds of this application in the preparation of a medicament for inhibiting GCN2 in cells. This application further includes one or more of the compounds of this application for inhibiting GCN2 in cells.

[0150] Since the compounds of this application have been shown to inhibit GCN2 protein activity, they can be used to treat diseases, disorders, or conditions by inhibiting GCN2. Therefore, the compounds of this application can be used as pharmaceuticals. Accordingly, this application includes the compounds of this application for use as pharmaceuticals.

[0151] Therefore, this application also includes methods for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more compounds of this application to an individual in need.

[0152] This application also includes the use of one or more of the compounds of this application for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2, and the use of one or more of the compounds of this application in the preparation of a medicament for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2. This application further includes one or more of the compounds of this application for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2.

[0153] "GCN2" is a protein kinase belonging to the eukaryotic initiation factor 2α (eIF2α) kinase family. In some embodiments, this serine / threonine protein kinase is an enzyme encoded by human GCN2 or EIF2AK4 (gene ID: 851877), which contains... Mol. Cell. Biol. 1995 , The amino acid sequence disclosed in 15 (8): 4497-506.

[0154] In some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is a neoplastic condition. Therefore, this application also includes a method of treating a neoplastic condition, comprising administering a therapeutically effective amount of one or more of the compounds of this application to an individual in need. This application also includes the use of one or more of the compounds of this application for treating a neoplastic condition, and the use of one or more of the compounds of this application in the preparation of a medicament for treating a neoplastic condition. This application further includes one or more of the compounds of this application for treating a neoplastic condition. In some embodiments, the therapeutic amount effectively improves at least one symptom of the neoplastic condition, for example, by reducing cell proliferation or tumor mass in an individual in need of such treatment.

[0155] Tumors can be benign (such as uterine fibroids and melanocytic nevi), potentially malignant (such as carcinoma in situ), or malignant (i.e., cancer). Exemplary neoplastic conditions include so-called solid tumors and liquid tumors, including but not limited to carcinomas, sarcomas, metastatic conditions (e.g., tumors originating from the prostate), hematopoietic system neoplastic conditions (e.g., leukemia, lymphoma, myeloma, and other malignant plasma cell diseases), metastatic tumors, and other cancers. Common cancers include breast cancer, prostate cancer, colon cancer, lung cancer, liver cancer, brain cancer, ovarian cancer, and pancreatic cancer.

[0156] The compounds of this application have been shown to inhibit the growth of cancer cells. In some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer.

[0157] Therefore, this application also includes a method of treating cancer, comprising administering a therapeutically effective amount of one or more of the compounds of this application to an individual in need. This application also includes the use of one or more of the compounds of this application for treating cancer, and the use of one or more of the compounds of this application in the preparation of a medicament for treating cancer. This application further includes one or more of the compounds of this application for treating cancer. In one embodiment, the compound is administered for cancer prevention in individuals with a predisposition to cancer, such as mammals.

[0158] In some implementations, the cancer is selected from, but not limited to: adult acute lymphoblastic leukemia; childhood acute lymphoblastic leukemia; adult acute myeloid leukemia; adrenocortical carcinoma; childhood adrenocortical carcinoma; AIDS-related lymphoma; AIDS-related malignancies; anal cancer; childhood cerebellar astrocytoma; childhood cerebral astrocytoma; extrahepatic bile duct carcinoma; bladder cancer; childhood bladder cancer; bone cancer, osteosarcoma / malignant fibrous histiocytoma; childhood brainstem glioma; adult brain tumors; childhood brain tumors, brainstem gliomas; childhood brain tumors. Tumors, cerebellar astrocytoma; Pediatric brain tumors, cerebral astrocytoma / malignant glioma; Pediatric brain tumors, ependymoma; Pediatric brain tumors, medulloblastoma; Pediatric brain tumors, supratentorial primitive neuroectodermal tumors; Pediatric brain tumors, visual pathway and hypothalamic gliomas; Pediatric brain tumors (other); Breast cancer; Breast cancer and pregnancy; Pediatric breast cancer; Male breast cancer; Pediatric bronchial adenoma / carcinoid; Pediatric carcinoid tumors; Gastrointestinal carcinoid tumors; Adrenocortical carcinoma; Islet cell carcinoma; Carcinoma of Unknown Primary Occurrence Primary; Primary central nervous system lymphoma; Childhood cerebellar astrocytoma; Childhood cerebral astrocytoma / malignant glioma; Cervical cancer; Childhood cancer; Chronic lymphocytic leukemia; Chronic myeloid leukemia; Chronic myeloproliferative disorders; Tenosynovial clear cell sarcoma; Colon cancer; Childhood colorectal cancer; Cutaneous T-cell lymphoma; Endometrial cancer; Childhood ependymoma; Ovarian epithelial cancer; Esophageal cancer; Childhood esophageal cancer; Ewing's family tumors; Childhood extracranial germ cell tumors; Gonadal extragerminal tumors; Extrahepatic bile duct cancer; Ocular cancer, intraocular melanoma; Ocular cancer, retinoblastoma; Gallbladder cancer; Gastric cancer; Childhood gastric cancer; Gastrointestinal carcinoid tumors; Childhood extracranial germ cell tumors; Gonadal extragerminal tumors; Ovarian germ cell tumors; Gestational trophoblastic tumors; Childhood brainstem glioma; Childhood visual pathway and hypothalamic glioma; Hairy cell leukemia; Head and neck cancer; Adult hepatocellular carcinoma (hepatitis) (primary); Childhood hepatocellular carcinoma (hepatitis) Primary Hodgkin's lymphoma; adult Hodgkin's lymphoma; childhood Hodgkin's lymphoma; gestational Hodgkin's lymphoma; hypopharyngeal cancer; pediatric hypothalamic and visual pathway glioma; intraocular melanoma; islet cell carcinoma (endocrine pancreas); Kaposi's sarcoma; renal cell carcinoma; laryngeal cancer; childhood laryngeal cancer; adult acute lymphoblastic leukemia; childhood acute lymphoblastic leukemia; adult acute myeloid leukemia; childhood acute myeloid leukemia; chronic lymphoblastic leukemia; chronic myeloid leukemia; hairy cell leukemia. Blood disorders; lip and oral cancer; primary liver cancer in adults; primary liver cancer in children; non-small cell lung cancer; small cell lung cancer; acute lymphoblastic leukemia in adults; acute lymphoblastic leukemia in children; chronic lymphocytic leukemia; AIDS-related lymphoma; primary central nervous system lymphoma; cutaneous T-cell lymphoma; Hodgkin lymphoma in adults; Hodgkin lymphoma in children; Hodgkin lymphoma during pregnancy; non-Hodgkin lymphoma in adults; non-Hodgkin lymphoma in children;Non-Hodgkin's lymphoma during pregnancy; primary central nervous system lymphoma; Waldenström macroglobulinemia; male breast cancer; adult malignant mesothelioma; childhood malignant mesothelioma; malignant thymoma; childhood medulloblastoma; melanoma; intraocular melanoma; Merkel cell carcinoma; malignant mesothelioma; metastatic squamous cell carcinoma of unknown primary origin (Metastatic Squamous Neck Cancer with Occult Primary); multiple endocrine neoplasia syndrome in children; multiple myeloma / plasma cell tumor; mycosis fungoides; myelodysplastic syndrome; chronic myeloid leukemia; acute myeloid leukemia in children; multiple myeloma; chronic myeloproliferative disorders; nasal cavity and paranasal sinus carcinoma; nasopharyngeal carcinoma; nasopharyngeal carcinoma in children; neuroblastoma; non-Hodgkin lymphoma in adults; non-Hodgkin lymphoma in children; non-Hodgkin lymphoma during pregnancy; non-small cell lung cancer; oral cancer in children; oral and lip cancer; oropharyngeal cancer; osteosarcoma / malignant fibrous histiocytoma of bone; ovarian cancer in children; ovarian epithelial carcinoma; ovarian germ cell tumors; ovarian low-malignant potential tumors. Tumors; Pancreatic cancer; Pediatric pancreatic cancer; Islet cell pancreatic cancer; Paranasal sinus and nasal cavity cancer; Parathyroid cancer; Penile cancer; Pheochromocytoma; Pediatric pineal and supratentorial primitive neuroectodermal tumors; Pituitary tumors; Plasma cell tumors / multiple myeloma; Pleural pleuroblastoma; Pregnancy and breast cancer; Pregnancy and Hodgkin lymphoma; Pregnancy and non-Hodgkin lymphoma; Primary central nervous system lymphoma; Adult primary liver cancer; Pediatric primary liver cancer; Prostate cancer; Rectal cancer; Renal cell (kidney) cancer; Pediatric renal cell carcinoma; Transitional cell carcinoma of the renal pelvis and ureter; Retinoblastoma; Pediatric rhabdomyosarcoma; Salivary gland cancer; Pediatric salivary gland cancer; Sarcoma, Ewing's Family of Tumors; Kaposi's sarcoma; osteosarcoma / malignant fibrous histiocytoma of bone; pediatric sarcoma, rhabdomyosarcoma; adult soft tissue sarcoma; pediatric soft tissue sarcoma; Cézari syndrome; skin cancer; pediatric skin cancer; skin cancer (melanoma); Merkel cell skin cancer; small cell lung cancer; small bowel cancer; adult soft tissue sarcoma; pediatric soft tissue sarcoma; metastatic squamous cell carcinoma of the neck of unknown primary origin; gastric cancer; pediatric gastric cancer; pediatric supratentorial primitive neuroectodermal tumor; cutaneous T-cell lymphoma; testicular cancer; pediatric thymoma; malignant thymoma; thyroid cancer; pediatric thyroid cancer; transitional cell carcinoma of the renal pelvis and ureter; gestational trophoblastic tumor; pediatric cancer of unknown primary origin; rare pediatric cancer; transitional cell carcinoma of the ureter and renal pelvis; urethral cancer; uterine sarcoma; vaginal cancer; pediatric visual pathway and hypothalamic glioma; vulvar cancer; Waldenström macroglobulinemia; and nephroblastoma. Metastasis of the above cancers can also be treated according to the methods described in this article.

[0159] In some embodiments, the cancer is any cancer in which cells exhibit increased expression or activation of one or more genes encoding GCN2 under stress conditions. "Increased expression" means any increase in the expression of one or more genes encoding GCN2 in a cell compared to the expression of one or more genes encoding GCN2 in corresponding normal or healthy cells.

[0160] In some embodiments, the cancer is selected from one or more of solid tumors, breast cancer, colon cancer, bladder cancer, skin cancer, head and neck cancer, liver cancer, lung cancer, pancreatic cancer, ovarian cancer, prostate cancer, bone cancer, and glioblastoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is skin cancer. In some embodiments, the cancer is head and neck cancer. In some embodiments, the cancer is colorectal cancer (CRC). In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is glioblastoma. In some embodiments, the bone cancer is osteosarcoma.

[0161] In some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is a disease, disorder, or condition associated with uncontrolled and / or abnormal cell activity directly or indirectly affected by inhibiting GCN2. In another embodiment, the uncontrolled and / or abnormal cell activity directly or indirectly affected by inhibiting GCN2 is cell proliferative activity.

[0162] Therefore, this application also includes a method for inhibiting cell proliferation activity, comprising applying an effective amount of one or more compounds of this application to the cells. This application also includes the use of one or more compounds of this application for inhibiting cell proliferation activity, and the use of one or more compounds of this application in the preparation of a medicament for inhibiting cell proliferation activity. This application also includes one or more compounds of this application for inhibiting cell proliferation activity.

[0163] This application also includes a method for inhibiting uncontrolled and / or abnormal cell activity directly or indirectly affected by inhibiting GCN2 in cells of a biological sample or an individual, the method comprising applying an effective amount of one or more compounds of this application to the cells. This application also includes the use of one or more compounds of this application for inhibiting uncontrolled and / or abnormal cell activity directly or indirectly affected by inhibiting GCN2 in cells, and the use of one or more compounds of this application in the preparation of a medicament for inhibiting uncontrolled and / or abnormal cell activity directly or indirectly affected by inhibiting GCN2 in cells. This application also includes one or more compounds of this application for inhibiting uncontrolled and / or abnormal cell activity directly or indirectly affected by inhibiting GCN2 in cells.

[0164] In some embodiments, the disease, disorder, or symptom that can be treated by inhibiting GCN2 is peripheral neuropathy. Therefore, this application also includes a method of treating peripheral neuropathy, comprising administering a therapeutically effective amount of one or more of the compounds of this application to an individual in need. This application also includes the use of one or more of the compounds of this application for treating peripheral neuropathy, and the use of one or more of the compounds of this application in the preparation of a medicament for treating peripheral neuropathy. This application further includes one or more of the compounds of this application for treating peripheral neuropathy.

[0165] In some embodiments, the peripheral neuropathy is Charco-Marie-Tuss (CMT) peripheral neuropathy. Heterozygous mutations in six genes encoding cytosine aminoacyl-tRNA synthetase (AARS) result in axonal and intermediate CMT peripheral neuropathy. AARS is a universally expressed enzyme that covalently links amino acids to its homologous tRNA (tRNA aminoacylation). Aminoacylated tRNA is used by ribosomes for mRNA translation. In Charco-Marie-Tuss (CMT) peripheral neuropathy, the mutated tRNA synthetase activates the integrated stress response (ISR) via the sensor kinase GCN2. Chronic activation of ISR leads to pathophysiology, while gene deletion or pharmacological inhibition of GCN2 alleviates the peripheral neuropathy. Therefore, in some implementations, the disease, disorder, or condition that can be treated by inhibiting GCN2 is Charcot-Marie-Tuss (CMT) peripheral neuropathy.

[0166] Therefore, this application also includes a method of treating Charcot-Marie-Tuss (CMT) peripheral neuropathy, comprising administering a therapeutically effective amount of one or more of the compounds of this application to an individual in need. This application also includes the use of one or more of the compounds of this application for treating Charcot-Marie-Tuss (CMT) peripheral neuropathy, and the use of one or more of the compounds of this application in the preparation of a medicament for treating Charcot-Marie-Tuss (CMT) peripheral neuropathy. This application further includes one or more of the compounds of this application for treating Charcot-Marie-Tuss (CMT) peripheral neuropathy.

[0167] This application also includes methods for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more of the compounds of this application in combination with another known pharmaceutical agent that can be used to treat diseases, disorders, or conditions that can be treated by inhibiting GCN2 to an individual in need. This application also includes the use of one or more of the compounds of this application in combination with another known pharmaceutical agent that can be used to treat diseases, disorders, or conditions that can be treated by inhibiting GCN2, and the use of one or more of the compounds of this application in combination with another known pharmaceutical agent that can be used to treat diseases, disorders, or conditions that can be treated by inhibiting GCN2 in the preparation of a medicament for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2. This application also includes one or more of the compounds of this application in combination with another known pharmaceutical agent that can be used to treat diseases, disorders, or conditions that can be treated by inhibiting GCN2 for the purpose of treating diseases, disorders, or conditions that can be treated by inhibiting GCN2.

[0168] In one embodiment, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer and / or peripheral neuropathy.

[0169] In some implementations, GCN2 is suppressed in the uses and methods of this application.

[0170] In one implementation, an “individual in need” is an individual suffering from the aforementioned disease, disorder, or condition that requires treatment.

[0171] In one embodiment, the individual is a mammal. In another embodiment, the individual is a human.

[0172] In some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer, and the one or more compounds of this application are administered or used in combination with one or more additional cancer treatments. In another embodiment, the one or more additional cancer treatments are selected from one or more radiotherapy treatments, chemotherapy treatments, targeted therapies such as antibody therapy (including anti-PD1 and / or anti-PD-L1 antibodies) and small molecule therapies such as tyrosine kinase inhibitor therapy, glutaminase inhibitors (e.g., glutaminase-1 (GLS1) inhibitors) and asparagine synthase (ASNS) inhibitors, immunotherapy, hormone therapy, and anti-angiogenic therapy.

[0173] In some embodiments, the chemotherapy is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is cisplatin. Therefore, in some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer, and one or more of the compounds of this application are administered or used in combination with cisplatin. In some embodiments, the chemotherapeutic agent is L-asparaginase (L-ASNase). Therefore, in some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer, and one or more of the compounds of this application are administered or used in combination with L-asparaginase (L-ASNase).

[0174] In some embodiments, the small molecule therapy is a glutaminase (e.g., glutaminase-1 (GLS1)) inhibitor or an asparagine synthase (ASNS) inhibitor. Therefore, in some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer, and the one or more compounds of this application are administered or used in combination with one or more glutaminase inhibitors (e.g., GLS1 inhibitors) and / or asparagine synthase (ASNS) inhibitors.

[0175] In some embodiments, the disease, disorder, or condition that can be treated by inhibiting GCN2 is cancer, and the one or more compounds of this application are administered or used in combination with one or more glutaminase inhibitors (e.g., GLS1 inhibitors) and / or asparagine synthase (ASNS) inhibitors and / or L-asparaginase (L-ASNase).

[0176] Asparagine deprivation via L-asparaginase (L-ASNase) is an effective cancer treatment strategy, but resistance develops due to the upregulation of asparagine synthase (ASNS) (the only human enzyme that synthesizes asparagine). Annu. Rev. Biochem. 2006, 75 (1), 629-654. The efficacy of L-asparaginase in solid tumors is limited by dose-related toxicity. OncoTargets and Therapy2017, pp 1413-1422). Large-scale loss-of-function genetic in vitro screening identifies ASNS as a cancer-dependent factor in several solid malignancies. Cell 2017, 170 (3), 564-576.e16. Cell 2017, 170 (3), 577-592.e10). Genome-wide CRISPR screening has revealed that drug resistance mechanisms in cancer cells are mediated through the GCN2-ATF4 axis, which aims to restore amino acid levels to promote survival. Therefore, pharmacological inhibition of GCN2 synergizes with L-asparaginase-mediated asparagine deprivation in ASNS-deficient cells, suggesting novel potential therapeutic combinations in cancer treatment.

[0177] Therefore, in some embodiments, this application also includes a method for improving the efficacy of one or more cancer treatments for treating cancer, which includes administering an effective amount of one or more compounds of this application in combination with an effective amount of the one or more cancer treatments to an individual in need.

[0178] This application also includes the use of one or more of the compounds of this application in combination with one or more cancer treatments to enhance the efficacy of said one or more cancer treatments in treating cancer, and the use of one or more of the compounds of this application in combination with one or more cancer treatments to enhance the efficacy of said one or more cancer treatments in treating cancer. This application further includes one or more of the compounds of this application, in combination with one or more cancer treatments, for enhancing the efficacy of said one or more cancer treatments in treating cancer.

[0179] In some embodiments, the one or more cancer treatments are selected from one or more radiotherapy, chemotherapy, targeted therapies such as antibody therapy (including anti-PD1 and / or anti-PD-L1 antibodies) and small molecule therapies such as tyrosine kinase inhibitor therapy, glutaminase inhibitors (e.g., GLS1 inhibitors) and / or asparagine synthase (ASNS) inhibitors, immunotherapy, hormone therapy and anti-angiogenic therapy.

[0180] In some embodiments, the chemotherapy is a chemotherapeutic agent. In some embodiments, the chemotherapeutic agent is cisplatin. Therefore, in some embodiments, one or more of the compounds of this application are administered or used in combination with cisplatin to enhance the efficacy of cisplatin in treating cancer.

[0181] In some embodiments, the chemotherapeutic agent is L-asparaginase (L-ASNase). Therefore, in some embodiments, one or more of the compounds of this application are administered or used in combination with L-asparaginase (L-ASNase) to enhance the efficacy of L-ASNase in treating cancer.

[0182] In some embodiments, the small molecule therapy is a glutaminase inhibitor (e.g., a GLS1 inhibitor) or an asparagine synthase (ASNS) inhibitor. Therefore, in some embodiments, the one or more compounds of this application are administered or used in combination with one or more glutaminase inhibitors (e.g., GLS1 inhibitors) and / or one or more asparagine synthase (ASNS) inhibitors to enhance the efficacy of the one or more glutaminase inhibitors (e.g., GLS1 inhibitors) or the one or more ASNS inhibitors in treating cancer.

[0183] In some embodiments, the cancer is associated with low asparagine synthase (ASNS) expression. In some embodiments, the cancer is associated with low asparagine synthase (ASNS) expression, and the chemotherapeutic agent is L-asparaginase (L-ASNase). Therefore, in some embodiments, one or more of the compounds of this application are administered or used in combination with L-asparaginase (L-ASNase) to enhance the efficacy of L-ASNase in treating cancers associated with low asparagine synthase (ASNS) expression.

[0184] In some embodiments, the cancer is associated with overexpression or dysregulation of asparagine synthase (ASNS). In some embodiments, the cancer is associated with overexpression or dysregulation of asparagine synthase (ASNS), and the chemotherapeutic agent is one or more asparagine synthase (ASNS) inhibitors and / or L-asparaginase. Therefore, in some embodiments, the one or more compounds of this application are administered or used in combination with one or more asparagine synthase (ASNS) inhibitors and / or with L-asparaginase for the treatment of cancer associated with overexpression or dysregulation of asparagine synthase (ASNS). In some embodiments, the chemotherapeutic agent is one or more asparagine synthase (ASNS) inhibitors and L-asparaginase.

[0185] In some embodiments, the cancer is associated with low expression of asparaginase synthase (ASN) and low expression of glutaminase (e.g., GLS1). In some embodiments, the cancer is associated with low expression of asparaginase synthase (ASN) and low expression of glutaminase (e.g., GLS1), and the chemotherapeutic agent is an L-asparaginase (L-ASNase) and / or one or more glutaminase inhibitors. Therefore, in some embodiments, the one or more compounds of this application are administered or used in combination with L-asparaginase (L-ASNase) and / or one or more glutaminase inhibitors to enhance the efficacy of L-ASNase and / or the one or more glutaminase inhibitors in treating cancers associated with low expression of asparaginase synthase (ASN) and low expression of glutaminase (e.g., GLS1). In some embodiments, the glutaminase inhibitor is a GLS1 inhibitor. In some embodiments, the chemotherapeutic agent is an L-asparaginase (L-ASNase) and one or more glutaminase inhibitors.

[0186] In some embodiments, the cancer is associated with overexpression or dysregulation of asparaginase synthase (ASNS) and overexpression or dysregulation of glutaminase (e.g., GLS1). In some embodiments, the cancer is associated with overexpression or dysregulation of asparaginase synthase (ASNS) and overexpression or dysregulation of glutaminase (e.g., GLS1), and the chemotherapeutic agent is L-asparaginase (L-ASNase), one or more glutaminase inhibitors, and / or one or more asparaginase synthase (ASNS) inhibitors. Therefore, in some embodiments, the one or more compounds of this application are administered or used in combination with L-asparaginase (L-ASNase) and / or one or more glutaminase inhibitors and / or one or more asparaginase synthase (ASNS) inhibitors to enhance the efficacy of L-ASNase and / or the one or more glutaminase inhibitors and / or the one or more asparaginase synthase (ASNS) inhibitors in treating cancers associated with overexpression or dysregulation of asparaginase synthase (ASNS) and overexpression or dysregulation of glutaminase (e.g., GLS1). In some embodiments, the glutaminase inhibitor is a GLS1 inhibitor. In some embodiments, the chemotherapeutic agent is L-asparaginase (L-ASNase), one or more glutaminase inhibitors, and one or more asparagine synthase (ASNS) inhibitors.

[0187] The compounds of this application may be used alone or in combination with other known pharmaceutical agents for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2. When used in combination with other pharmaceutical agents for treating diseases, disorders, or conditions that can be treated by inhibiting GCN2, one embodiment is that the compounds of this application are administered simultaneously with those pharmaceutical agents. As used herein, “simultaneous administration” of two substances to an individual means providing each of the two substances such that they are simultaneously biologically active in the individual. Specific details of administration will depend on the pharmacokinetics of the two substances in the presence of each other and may include administration of the two substances within hours of each other, or even administration of one substance within 24 hours of administration of the other substance, if the pharmacokinetics are appropriate. The design of suitable administration regimens is conventional to those skilled in the art. In certain embodiments, the two substances will be administered substantially simultaneously (i.e., within minutes of each other) or as a single composition containing both substances. Another embodiment of this application is the administration of a combination of pharmaceutical agents to an individual in a non-simultaneous manner. In some embodiments, the compounds of this application are administered simultaneously or sequentially with another therapeutic agent in a separate unit dosage form or in a single unit dosage form. Therefore, this application provides a single-unit dosage form comprising one or more compounds of this application (e.g., compounds of Formula I), an additional therapeutic agent, and a pharmaceutically acceptable carrier.

[0188] Treatment methods include administering a therapeutically effective amount of one or more of the compounds of this application to an individual, and optionally consist of a single administration, or alternatively include a series of administrations, and optionally include concurrent administration or the use of one or more other therapeutic agents. For example, in some embodiments, the compounds of this application are administered at least once a week. In some embodiments, for a given treatment, the compound is administered to an individual about once every two or three weeks, or about once a week to about once a day. In another embodiment, the compound is administered 2, 3, 4, 5, or 6 times daily. The length of the treatment period depends on a variety of factors, such as the severity of the disease, disorder, or symptom, the individual's age, the concentration and / or activity of the compounds of this application, and / or combinations thereof. It should also be understood that the effective dose of the compound used for treatment may be increased or decreased during a particular treatment regimen. Changes in dose can be produced and become apparent by standard diagnostic assays known in the art. In some cases, chronic administration may be required. For example, the compound is administered to the individual in an amount and for a duration sufficient to treat the individual. In some embodiments, the treatment includes prophylactic treatment. For example, individuals with early-stage cancer can be treated to prevent progression, or alternatively, individuals in remission can be treated with the compounds or compositions of this application to prevent recurrence.

[0189] The dosage of the compounds in this application varies depending on many factors, such as the pharmacodynamic properties of the compound, the administration method, the recipient's age, health status and weight, the nature and severity of symptoms, the frequency of treatment and the type of concurrent treatment (if any), and the clearance rate of the compound in the individual being treated. Those skilled in the art can determine an appropriate dosage based on these factors. The compounds in this application may be initially administered at an appropriate dosage that can be adjusted as needed based on clinical response. The dosage is typically selected to maintain serum levels of the compounds in this application at about 0.01 µg / cc to about 1000 µg / cc, or about 0.1 µg / cc to about 100 µg / cc. As a representative example, the oral dosage range for one or more compounds in this application for adults is about 0.05 mg / day to about 1000 mg / day, suitably about 0.1 mg / day to about 500 mg / day, more suitably about 1 mg / day to about 200 mg / day. For parenteral administration, representative doses are about 0.001 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 1 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. For oral administration, representative doses are about 0.001 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.01 mg / kg to about 1 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. For suppository administration, representative doses are about 0.1 mg / kg to about 10 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. The compounds of this application can be administered in a single daily, weekly, or monthly dose, or the total daily dose can be divided into two, three, or four daily doses.

[0190] In one embodiment, the effective amount varies based on factors such as disease state, individual age, sex, and / or weight. In a further embodiment, the amount of a given one or more compounds corresponding to the effective amount may vary based on factors such as the given one or more drugs or compounds, pharmaceutical formulation, route of administration, condition, type of disease or disorder, and identity of the individual to be treated, but may be conventionally determined by those skilled in the art.

[0191] For clarity, in the foregoing, the term "compound" also includes embodiments in which one or more compounds are mentioned. Similarly, the term "compound of this application" also includes embodiments in which only one compound is mentioned.

[0192] V. Method for preparing the compounds of this application The compounds of this application can be prepared by various synthetic methods. The choice of specific structural features and / or substituents may affect the choice of one method relative to another. The choice of a specific method for preparing a given compound of formula I is within the scope of those skilled in the art. Some of the starting materials used to prepare the compounds of this application are available from commercial chemical sources. For example, other starting materials described below can be readily prepared from available precursors using simple transformations known in the art.

[0193] Compounds of Formula I can generally be prepared according to the methods shown in the following route. In the structural formulas shown below, unless otherwise indicated, the variables are as defined in Formula I. Those skilled in the art will understand that many of the reactions depicted in the following route are sensitive to oxygen and water, and will know that reactions should be carried out under anhydrous, inert atmospheres as needed. Those skilled in the art will understand that the reaction temperatures and times are presented for illustrative purposes only and can be varied to optimize yields.

[0194] Therefore, in some embodiments, the compound of formula I is prepared as shown in route 1.

[0195] Route 1 Commercially available 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (Hal = Br, X1 = C or N, formula A) is oxidized with a suitable oxidant such as m-chloroperoxybenzoic acid (m-CPBA) to a sulfoxide intermediate of formula B. Then, the compound of formula B is coupled with a suitable amine of formula C or its protected form in the presence of a suitable base such as Cs₂CO₃ or DIPEA and in a suitable solvent such as iPrOH, MeCN, or DMF to give an intermediate of formula D. Using suitable conditions such as the Suzuki-Miyaura coupling conditions, the intermediate is then oxidized with a borate ester or boric acid of formula G (where R...). a and R b Independently for C 1-6 Alkyl groups, or linked together with the B and O atoms therebetween, form a 4- to 6-membered saturated or unsaturated ring, which is optionally bounded by one or two C atoms. 1-3 Compounds of formula D are treated with alkyl substitution to obtain compounds of formula I.

[0196] Borate esters or borates of formula G are prepared, for example, by coupling an arylsulfonyl compound of formula E, wherein Y' is a halogen such as Cl, with a borate ester or borate aniline compound of formula F in the presence of a suitable base such as pyridine.

[0197] Route 2 In some embodiments, the compound of formula I is prepared as shown in route 2. Therefore, the intermediate compound of formula D, where Hal is a halogen such as Br, is coupled with a substituted 3-aniline borate ester compound of formula F (prepared from a commercially available or synthetic corresponding 3-haloaniline compound under standard boronylation conditions such as Miyaura borylation, using (pinacolyl)diboron under alkaline conditions with a suitable catalyst such as PdCl2(dppf) ([1,1'-bis(diphenylphosphino)ferrocene]palladium(II):CH2Cl2 complex)) under suitable coupling conditions such as Suzuki-Miyaura coupling conditions to give the intermediate compound of formula J. The compound of formula J is then sulfonated, for example, under alkaline conditions, with a heterocyclic sulfonyl halide compound of formula E (where Y' is a halogen) to give the compound of formula I.

[0198] Route 3 In some embodiments, the compound of formula I is prepared as shown in route 3. Therefore, the compound of formula A is borated under standard boration conditions such as Miyaura boration conditions, for example in the presence of suitable reagents such as diboronol, PdCl2dppf:CH2Cl2 complex and bases such as KOAc, in a suitable solvent such as dioxane, and at a suitable temperature such as 100-110°C, to obtain the borated compound of formula K, wherein R... c and R d Independently for C 1-6 Alkyl groups, or linked together with the B and O atoms therebetween, form a 4- to 6-membered saturated or unsaturated ring, which is optionally bounded by one or two C atoms. 1-3 Alkyl substitution. Subsequently, the compound of formula K is treated with various halosulfonamides of formula H, where Y'' is a halogen (prepared, for example, by coupling a sulfonyl halide of formula E (where Y' is a halogen) with an aniline compound of formula M under suitable conditions, such as the Suzuki-Miyaura coupling conditions), to obtain the compound of formula L. The compound of formula L is oxidized to the intermediate sulfoxide with a suitable oxidant such as m-chloroperoxybenzoic acid (m-CPBA), and then coupled with various suitable amine compounds of formula C under basic conditions to obtain the compound of formula I.

[0199] Route 4 In some embodiments, compounds of formula I are prepared as shown in route 4. Thus, under suitable coupling conditions, such as in the presence of a base (e.g., pyridine), a compound of formula N is coupled with a sulfonyl halide of formula E in which Y' is a halogen to obtain a compound of formula L. The compound of formula L is oxidized to the intermediate sulfoxide with a suitable oxidant such as m-chloroperoxybenzoic acid (m-CPBA), and then coupled with various suitable amine compounds of formula C under basic conditions to obtain a compound of formula I.

[0200] Route 5 In some embodiments, compounds of formula I are prepared as shown in route 5. Therefore, under suitable coupling conditions, such as the Suzuki-Miyaura coupling conditions, an intermediate compound of formula D, wherein Hal is a halogen such as Br, is coupled to a borate ester compound of formula G to obtain compounds of formula I.

[0201] Route 6 In some implementations, R 2 = H, compound of formula I is prepared as shown in route 6. Therefore, under acidic conditions, R in it is prepared... 2 It is the deprotection of intermediate compounds of formula O with suitable protecting groups such as p-methoxybenzyl (PMB) that yields R. 2 = H, a compound of formula I.

[0202] Route 7 In some implementations, such as route 7, X 1 =N compounds of formula N (compounds of formula N-1) are prepared from readily available 2,4-dichloro-5-nitropyrimidine (compounds of formula P). Nitro reduction is carried out under suitable conditions, such as in the presence of Fe, to give an amine compound of formula Q, which is then subjected to a suitable amine (e.g., R) in the presence of a base (e.g., DIPEA). 2 Substitution with NH2) yields the intermediate compound of formula R. Cyclation of the compound of formula R with ethyl glyoxylate, followed by substitution with sodium methanethiol, yields the intermediate methylthioyl compound of formula T. Radical-mediated coupling with tert-butyl nitrite yields the intermediate compound of formula V. Subsequent amination under suitable conditions, such as in the presence of diphenylmethylimine, Xantphos, Pd(OAc)2, and Cs2CO3, yields the compound of formula N-1.

[0203] Route 8 In some embodiments, as shown in Route 8, intermediate borate esters or boric acids of formula F are prepared from halogenated derivative compounds of formula W (where Hal' is a halogen, such as Br) under standard boration conditions such as Miyaura boration conditions, for example in the presence of suitable reagents such as bis(pinacol)diboron, PdCl2dppf:CH2Cl2 complex and bases such as potassium acetate (KOAc), in a suitable solvent such as dioxane, at a suitable temperature such as 100-110 °C.

[0204] Route 9 In some embodiments, as shown in route 9, the intermediate compound of formula D (compound of formula D-1), wherein Hal is Br, is prepared from a compound of formula Z, wherein Hal'' is a halogen such as Cl, by reacting it with various suitable amino compounds of formula C under suitable conditions, for example in the presence of a base (such as KOAc or DIPEA), to give a compound of formula AA. Subsequently, the compound of formula AA is brominated, for example with Br2 in a suitable solvent such as acetonitrile (MeCN), to give a compound of formula D-1.

[0205] Generally, the above reactions are carried out in a suitable inert organic solvent and at temperature and time conditions optimized for the yield of the desired compound. Examples of suitable inert organic solvents include, but are not limited to, 2-propanol, dimethylformamide (DMF), 1,4-dioxane, dichloromethane (DCM), chloroform, tetrahydrofuran (THF), toluene, etc.

[0206] Salts of the compounds in this application are typically formed by dissolving a neutral compound in an inert organic solvent and adding the desired acid or base, and then separating the resulting salt by filtration or other known means.

[0207] The formation of the desired compound salt is achieved using standard techniques. For example, a neutral compound is treated with an acid or base in a suitable solvent, and the resulting salt is separated by filtration, extraction, or any other suitable method.

[0208] The formation of solvates varies depending on the compound and the solvate. Generally, solvates are formed by dissolving the compound in a suitable solvent and separating the solvate by cooling or using an antisolvent. Solvates are typically dried or azeotropic under ambient conditions. Choosing suitable conditions to form a particular solvate is a skill in the art. Examples of suitable solvents are ethanol, water, etc. When water is used as the solvent, the molecule is called a "hydrate". The formation of solvates of the compounds in this application varies depending on the compound and the solvate. Generally, solvates are formed by dissolving the compound in a suitable solvent and separating the solvate by cooling or using an antisolvent. Solvates are typically dried or azeotropic under ambient conditions. Choosing suitable conditions to form a particular solvate is a skill in the art.

[0209] The prodrugs of the compounds in this application may be conventional esters, for example, formed with available hydroxyl, thiol, amino, or carboxyl groups. For example, the available hydroxyl or amino groups may be acylated using an activated acid in the presence of a base and optionally in an inert solvent (e.g., an acyl chloride in pyridine).

[0210] Throughout the methods described herein, it should be understood that, where appropriate, suitable protecting groups will be added to and subsequently removed from various reactants and intermediates in a manner readily understood by those skilled in the art. Conventional procedures using such protecting groups and examples of suitable protecting groups are described, for example, in… Protective Groups in Organic Synthesis ", TW Green, PGM Wuts, Wiley-Interscience, New York, (1999). It should also be understood that the conversion of a group or substituent into another group or substituent by chemical manipulation can be carried out on any intermediate or final product along the synthetic pathway toward the final product, wherein the possible types of conversion are limited only by the inherent incompatibility of the other functional groups carried by the molecule at that stage with the conditions or reagents used for the conversion. Such inherent incompatibility, and the ways to circumvent them by performing appropriate conversion and synthetic steps in the appropriate order, are readily understood by those skilled in the art. Examples of conversions are given herein, and it should be understood that the conversions described are not limited to the general groups or substituents that have been exemplified. References and descriptions of other suitable conversions can be found in "Comprehensive Organic Transformations - A Guide to Functional Group Preparations" RC Larock, VHC Publishers, Inc. (1989). References and descriptions of other suitable reactions can be found in organic chemistry textbooks, such as, " Advanced Organic Chemistry", March, 4th ed. McGraw Hill (1992) or" Organic Synthesis Smith, McGraw Hill, (1994). Purification techniques for intermediates and end products include, for example, normal and reversed-phase chromatography on a column or rotating disk, recrystallization, distillation, and liquid-liquid or solid-liquid extraction, which will be readily understood by those skilled in the art.

[0211] The product of the method described in this application can be separated by known methods, such as solvent evaporation, filtration, centrifugation, chromatography, or other suitable methods.

[0212] Those skilled in the art will recognize that when the reaction steps of this application are carried out in a variety of solvents or solvent systems, the reaction steps can also be carried out in a mixture of suitable solvents or solvent systems.

[0213] Example The following non-limiting embodiments illustrate this application: A. Synthesis and Characterization of Exemplary Compounds in this Application General Method General method MB (Miyaura borylation) In a microwave reactor (typically 90-100 °C) or an oil bath (typically 100-110 °C), a degassed mixture of aryl halides (1.0 equivalent), B2pin2 (1.3 equivalent), KOAc (3.5 equivalent), and typically PdCl2(dppf)*CH2Cl2 or PdCl2(dppf) (0.1 equivalent) of 1,4-dioxane is heated under argon atmosphere in a sealed environment. Subsequently, the crude mixture is typically used directly for the next step of the Suzuki-Miyaura cross-coupling reaction without further purification.

[0214] The general method is SMC (Suzuki Miyaura cross-coupling). Add arylboronic acid or arylboronic ester (usually 1-1.5 equivalents, arylboronic esters are often used as a crude mixture in 1,4-dioxane) to a vial equipped with a stir bar and filled with Ar or N2, a base (Cs2CO3, usually 3 equivalents), an aryl halide (usually 1 equivalent), and a catalyst / ligand (most commonly one of PdCl2(dppf)*CH2Cl2 or PdCl2(dppf); usually 0.1 equivalent). Seal the vial, add H2O and an organic solvent or mixture of organic solvents (DME or 1,4-dioxane). Degas the reaction mixture with argon or N2 by repeatedly evacuating and backfilling with an inert gas, then seal and heat in a microwave reactor or oil bath for the specified time. After LCMS analysis confirms the reaction is complete, the mixture is concentrated under reduced pressure and deposited onto a diatomaceous earth (Celite) short column or SiO2 sample tube. The mixture is then analyzed by rapid chromatography (typically using SiO2InnoFlash@column, SiO2Biotage@column, or SiO2RediSep@Rf column and hexane-EtOAc, CH2Cl2-MeOH, CH2Cl2-MeOH-NH3, or CH2Cl2-MeOH-concentrated aq NH3 (89:9:1 v / v / v) in CH2Cl2) or preparative HPLC (typically using Biotage@SNAP KP-C). 18 -HS column or RedisSep@Rf C 18 Purify with MeOH in H2O + 0.05% TFA or MeCN in H2O + 0.1% formic acid, optionally followed by filtration through a Waters PoraPak™ CX column or an Isolute™ SCX-2 column, wash with MeOH and elute the desired substance with a MeOH solution of 2 M NH3.

[0215] General method NS (N-sulfonation) A solution of substituted 3-bromoaniline (1 equivalent) and anhydrous pyridine (typically 1.5–2.0 equivalents) in CH₂Cl₂ (typically 0.06–0.19 M) is treated at 0 °C with a fraction of solid arylsulfonyl chloride (typically 1 equivalent). The reaction is allowed to slowly reach room temperature and stirred overnight. The reaction mixture is then concentrated under reduced pressure onto diatomaceous earth, or the organic extract is concentrated under reduced pressure by extraction and washing with H₂O, deposited onto Biotage@ sample tubes or diatomaceous earth, and purified by silica gel rapid chromatography using one of the following columns: InnoFlash@, Biotage@, or RediSep@Rf.

[0216] General method MO (mCPBA-mediated oxidation) At 0 °C, solid 3-chloroperoxybenzoic acid (mCPBA) (typically 1.1–2.2 equivalents, industrial grade, <77%) is added to a suspension of methylthioheteroaryl (1 equivalent) in CH₂Cl₂ or in a small amount of CH₂Cl₂. The reaction is then slowly heated to room temperature and stirred at room temperature until complete. The resulting mixture of sulfoxide and sulfone intermediates is washed by extraction with a saturated (satd) aqueous solution of NaHCO₃, followed by concentration of the organic layer or simple concentration under reduced pressure. The crude product is used directly without post-treatment or further purification.

[0217] intermediate Intermediate 1: N-(3-bromo-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide 3-Bromo-2,4-difluoroaniline (15.5 g, 74.3 mmol) was added to a stirred solution of 5-chloro-2-methoxypyridine-3-sulfonyl chloride (20 g, 83 mmol) in pyridine (200 mL), and the resulting reaction mixture was stirred at room temperature for 1 hour. Afterward, the reaction mixture was quenched with 2 M HCl aqueous solution (500 mL) and extracted with EtOAc (3 x 300 mL). The combined organic layers were washed with brine (3 x 300 mL), dried (Na₂SO₄), and concentrated under reduced pressure. The crude product was ground with a 2% EtOAc solution in hexane to give N-(3-bromo-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide as a white solid (17 g, 50%). MS (ESI) m / z [M+H] + 412.8 / 414.8. 1 H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H),8.53 (d, J = 2.4 Hz, 1H), 8.07 (d, J = 2.6 Hz, 1H), 7.37-7.24 (m, 2H), 3.92(s, 3H).

[0218] Intermediate 2: ** 6-Bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one Step 1: 6-Bromo-8-methyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one Under N2, K2CO3 (7.6 g, 55 mmol) was added to a cooled DMF (150 mL) solution of 10 g, 37 mmol of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one at -5 °C. The reaction mixture was stirred at -5 °C for 10 min, and then MeI (6.26 g, 44.1 mmol) was added at -5 °C. Stirring was continued at cooling for 45 min. The reaction mixture was then diluted with ice-cold H2O (150 mL), filtered, washed with hexane, and dried under vacuum to give a white solid of 6-bromo-8-methyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (4 g, 38 %). MS (ESI) m / z [M+H] + 286.0 / 288.0.

[0219] Step 2: 6-Bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one Under N2 conditions, m-CPBA (7.3 g, 41.9 mmol) was added fractionally to a cooled CH2Cl2 (45 mL) solution of 6-bromo-8-methyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (3 g, 10.5 mmol) at 0 °C. The reaction mixture was then stirred at room temperature for 16 h, diluted with a saturated aqueous solution of NaHCO3 (150 mL), and extracted with CH2Cl2 (3 x 60 mL). The combined organic layers were dried (anhydrous Na2SO4) and concentrated under reduced pressure. The mixture was ground with a 20% EtOAc solution in hexane to give a light brown solid of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (2.4 g, 72%). MS (ESI) m / z [M+H] + 318.0 / 320.0. 1 H NMR (400 MHz, DMSO- d6 ) δ 3.51 (s, 3H), 3.74 (s, 3H), 8.75 (s, 1H), 9.29 (s, 1H).

[0220] Intermediate 3: 6-(5-amino-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one Step 1: 2,4-Dichloropyrimidine-5-amine Fe powder (2.89 g, 51.5 mmol) was added to a stirred solution of 2,4-dichloro-5-nitropyrimidine (5 g, 26 mmol) in AcOH (50 mL) at 0 °C, and the reaction mixture was stirred at room temperature for 5 hours. The reaction mixture was filtered through diatomaceous earth, and the filter cake was washed with EtOAc (300 mL). The filtrate was concentrated under reduced pressure. The solid residue was ground with hexane to give an orange solid of 2,4-dichloropyrimidine-5-amine (3 g, 71%). MS (ESI) m / z [M+H] + 164.0.

[0221] Step 2: 2-Chloro-N 4 -Methylpyrimidine-4,5-diamine DIPEA (6.3 mL, 36 mmol) was added to a stirred solution of 2,4-dichloropyrimidin-5-amine (3 g, 18 mmol) in EtOH (30 mL) at room temperature. The reaction mixture was then stirred for 10 minutes, followed by the dropwise addition of MeNH2 (2 M THF solution, 19 mL, 37 mmol). The reaction mixture was then stirred at 80 °C for 16 hours. The reaction mixture was subsequently concentrated under reduced pressure. The crude product was ground with methyl tert-butyl ether to give a deep purple semi-solid 2-chloro-N 4 4,5-Methylpyrimidine-4,5-diamine (2.8 g, 96%). MS (ESI) m / z [M+H] + 159.04.

[0222] Step 3: 2-Chloro-8-methylpterin-7(8H)-one To the stirring 2-chloro-N 4 To a solution of 4,5-methylpyrimidine-4,5-diamine (11 g, 70 mmol) in EtOH (110 mL), a PhMe solution of ethyl glyoxylate (30%, 7.07 mL, 69.6 mmol) was added dropwise. The reaction mixture was stirred at 80 °C for 16 hours. After completion, the reaction mixture was concentrated under reduced pressure. The crude product was ground with methyl tert-butyl ether to give a white solid 2-chloro-8-methylpteridin-7(8H)-one (4.5 g, 33%). MS (ESI) m / z [M+H] + 197.0.

[0223] Step 4: 8-Methyl-2-(meththio)pterin-7(8H)-one MeSNa (0.91 g, 13 mmol) was added fractionally to a stirred solution of 2-chloro-8-methylpteridine-7(8H)-one (1.6 g, 8.1 mmol) in THF (16 mL). The reaction mixture was stirred at 80 °C for 16 h, then diluted with water (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were dried (anhydrous Na₂SO₄) and concentrated under reduced pressure. The crude substance was purified by rapid chromatography using a hexane solution of EtOAc to give a white solid, 8-methyl-2-(methylthio)pteridine-7(8H)-one (0.9 g, 53%). 1 H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.22 (s,1H), 3.54 (s, 3H), 2.63 (s, 3H).

[0224] Step 5: 6-(5-bromo-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one Under N2, tert-butyl nitrite (1.48 g, 14.3 mmol) was added to a solution of 5-bromo-2-fluoroaniline (1.8 g, 9.5 mmol) in MeCN (50 mL) at 0 °C. The resulting solution was stirred at 0 °C for 30 min. Subsequently, a solution of 8-methyl-2-(meththio)pteridine-7(8H)-one (0.5 g, 2.4 mmol) in MeCN (5 mL) was added at 0 °C. The reaction mixture was then stirred at room temperature for 16 h. The reaction mixture was then diluted with water (100 mL) and extracted with CH2Cl2 (3 x 100 mL). The combined organic layers were dried (anhydrous Na2SO4) and concentrated under reduced pressure. 6-(5-bromo-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one (0.2 g, 22%), an off-white solid, was obtained by rapid chromatographic purification using a hexane solution of EtOAc. MS (ESI) m / z [M+H] + 381.1 / 383.1.

[0225] Step 6: 6-(5-amino-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one To a solution of 1,4-dioxane (11 mL) of 6-(5-bromo-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one (1.1 g, 2.9 mmol), diphenylmethyleneimine (0.57 g, 3.2 mmol), Xantphos (0.5 g, 0.86 mmol), Pd(OAc)₂ (0.097 g, 0.43 mmol), and Cs₂CO₃ (2.8 g, 8.7 mmol) were added under N₂. The reaction mixture was degassed with N₂ for 10 min and stirred at 100 °C for 3 h. After completion, the reaction mixture was acidified with 2 M HCl aqueous solution (to pH=2) and extracted with EtOAc (3 x 100 mL). The organic layer was discarded, the aqueous layer was alkalized with 2 M NaOH aqueous solution and extracted with EtOAc (3 x 100 mL). The combined organic layers were dried (using anhydrous Na₂SO₄) and concentrated under reduced pressure. The crude substance was purified by column chromatography using 60-120 mesh silica gel and a hexane solution of EtOAc to give an orange solid, 6-(5-amino-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one (0.8 g, 87%). MS (ESI) m / z [M+H] + 318.2. 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (s, 1H), 6.98 (t, J = 8.6 Hz, 1H), 6.60-6.85 (m,2H), 5.31 (br.s., 2H), 3.62 (s, 3H), 2.65(s, 3H).

[0226] Intermediate 4: 6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpteridine-7(8H)-one Step 1: 2-((1R,4R)-4-(dimethylamino)cyclohexyl)amino)-8-methylpteridine-7(8H)-one To a stirred solution of 1.7 g (88 mmol) of 2-chloro-8-methylpteridine-7(8H)-one in DMF (17 mL), trans-N,N1-dimethylcyclohexane-1,4-diamine (1.69 g, 95.4 mmol) and K₂CO₃ (3.5 g, 26 mmol) were added at room temperature. Stirring was continued at room temperature for 16 hours. Afterward, the reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (25 mL), dried (anhydrous Na₂SO₄), and concentrated under reduced pressure. The crude product was purified by column chromatography using a solution of MeOH in CH₂Cl₂ to give a brown solid, 2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpteridine-7(8H)-one (0.83 g, 32%). MS (ESI) m / z [M+H] + 303.2.

[0227] Step 2: 6-Bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpteridine-7(8H)-one *HBr To a solution of 2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpteridine-7(8H)-one (0.5 g, 1.6 mmol) in MeCN (5.0 mL), Br2 (0.5 mL, 12 mmol) was added at 0 °C. The reaction mixture was stirred at 0 °C for 2 hours. After completion, the reaction mixture was directly injected into a silica gel column and purified by column chromatography using a solution of MeOH in CH2Cl2. The mixture was ground with n-pentane and Et2O to give a brown solid, 6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpteridine-7(8H)-one*HBr (0.467 g, 73 %). MS (ESI) m / z [M+H] + 381.3 / 383.2. 1 H NMR (400MHz, DMSO-d6) δ 9.64 (br.s., 1H), 8.50-8.80 (m, 1H), 8.30-8.00 (m, 1H), 3.70-3.40 (m, 3H)3.30-3.05 (m, 2H), 2.74 (s, 6H), 2.25-1.85 (m, 4H), 1.75-1.20 (s, 4H).

[0228] Intermediate 5: 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl)-2-methoxypyridine-3-sulfonamide and (5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid 4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (4.0 g, 17 mmol) was added to a stirred solution of 5-chloro-2-methoxypyridine-3-sulfonyl chloride (4.0 g, 17 mmol) in pyridine (40 mL). The resulting reaction mixture was stirred at room temperature for 1 hour. After completion, the reaction mixture was directly loaded onto silica gel and purified by column chromatography using a solution of MeOH in CH2Cl2. Grinding with CH₂Cl₂ and Et₂O yielded a mixture of 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl)-2-methoxypyridine-3-sulfonamide and (5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid as a white solid (1.7 g, 23%). MS (ESI) m / z [M+H] + 361.2.

[0229] Intermediate 6: (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid A solution of 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)aniline (2.5 g, 10.5 mmol) in anhydrous pyridine (30 mL) was added at 0 °C with 5-chloro-2-methoxypyridine-3-sulfonyl chloride (2.5 g, 10.3 mmol), and the reaction mixture was stirred at room temperature for 16 hours. Afterward, the reaction mixture was directly loaded onto a silica gel column and purified using a solution of MeOH in CH₂Cl₂ to give (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid as a brown solid (2.8 g, 74%). MS (ESI) m / z [MH] - 359.3.

[0230] Intermediate 7: 6-Bromo-8-methyl-2-(methylamino)pterin-7(8H)-one Step 1: 8-Methyl-2-(methylamino)pterin-7(8H)-one A solution of 2-chloro-8-methylpteridine-7(8H)-one (0.9 g, 4.57 mmol) in EtOH (9 mL) and DIPEA (1.1 g, 8.5 mmol) was stirred at room temperature for 15 minutes, followed by dropwise addition of MeNH2 (2 M THF solution, 2.5 mL, 5.0 mmol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours, then filtered, washed with EtOAc, and dried under vacuum to give a white solid of 8-methyl-2-(methylamino)pteridine-7(8H)-one (0.4 g, 46%). MS (ESI) m / z [M+H] + 192.19.

[0231] Step 2: 6-Bromo-8-methyl-2-(methylamino)pterin-7(8H)-one Br2 (4 g, 25.1 mmol) was added dropwise to a stirred solution of 8-methyl-2-(methylamino)pterin-7(8H)-one (0.8 g, 4.2 mmol) in MeCN (16 mL) at 0 °C. Stirring was continued at this temperature for 2 hours. After completion, the reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were dried (anhydrous Na2SO4) and concentrated under reduced pressure. The crude product was purified by preparative HPLC to give 6-bromo-8-methyl-2-(methylamino)pterin-7(8H)-one (0.15 g, 13%) as a pale yellow solid. 1 H NMR (400 MHz, DMSO- d6 ) δ 8.75-8.58 (m, 1H), 8.15-7.95 (m, 1H), 3.62-3.45 (m, 3H), 2.89 (d, J= 4.8 Hz, 3H). MS (ESI)m / z [M+H] + 270.2 / 272.2.

[0232] Intermediate 8: 2,4-Difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)aniline A solution of 3-bromo-2,4-difluoroaniline (15 g, 72.1 mmol), Pd(dppf)Cl2 (5.27 g, 7.21 mmol), B2pin2 (27.4 g, 108.1 mmol), and KOAc (21.2 g, 216.3 mmol) in 1,4-dioxane (275 mL) was stirred at 100 °C for 16 hours under N2. After completion, the solid was removed by filtration and washed with CH2Cl2 (150 mL). The filtrate was concentrated under reduced pressure and purified by column chromatography using a hexane (Hex) solution of EtOAc to give 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline as a white solid (10 g, 54%). MS (ESI) m / z [M+H] + 256.2 and 174.0 (corresponding boric acid). 1 H NMR (400 MHz, DMSO- d6 ) δ 6.83-6.79 (m, 1H),6.71 (t, J = 8.6 Hz, 1H), 4.91 (s, 2H), 1.29 (s, 12H).

[0233] Exemplary compounds of this application Example 1: N-(3-(2-amino-8-isopropyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-1) Step 1: 6-Bromo-8-isopropyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one A capped, dry microwave-safe vial containing 0.5 g (1.8 mmol) of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one and NaH (60% mineral oil, 0.110 g, 2.8 mmol) was evacuated and purged with argon. Anhydrous DMF (10 mL) was added, and the mixture was stirred at room temperature for 10 minutes. The reaction mixture was then heated at 50 °C for 30 minutes. i-PrI (0.276 mL, 2.76 mmol) was added in a single batch at room temperature. After brief stirring at room temperature, the reaction mixture was placed in an oil bath at 40 °C for 1.5 hours, and then allowed to stand at room temperature for 3 days and 18 hours. DMF was removed under reduced pressure, and the mixture was suspended in EtOH. H₂O was added, and the solid was collected by filtration. The filter cake was washed with H2O and vacuum dried to give a white solid, 6-bromo-8-isopropyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (565 mg, 75% yield based on 77% purity). MS (ESI) m / z [M+H] + 314.05 / 316.01.

[0234] Step 2: (8-Isopropyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid The reaction was prepared using the universal method MB, with 6-bromo-8-isopropyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.49 mmol, 77% purity), B2pin2 (162 mg, 0.64 mmol), KOAc (168 mg, 1.72 mmol), and PdCl2dppf•CH2Cl2 (40.0 mg, 0.049 mmol) in anhydrous 1,4-dioxane (12 mL), by sealed heating in an oil bath at 100 °C for 2 hours. The reaction was then left to stand overnight at room temperature. The sealed reaction was then heated in an oil bath at 120 °C for 3.5 hours. The crude mixture was used directly for the next step. MS (ESI) m / z [M+H] + 280.28.

[0235] Step 3: 5-Chloro-N-(2,4-difluoro-3-(8-isopropyl-2-(methylthio)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide The reaction mixture was prepared using the general method SMC, with Cs₂CO₃ (177 mg, 0.543 mmol), N-(3-bromo-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (100 mg, 0.242 mmol), (8-isopropyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid (137 mg, 0.152 mmol) (in 12 mL dioxane, 31%), and PdCl₂dppf•CH₂Cl₂ (14.79 mg, 0.018 mmol) and H₂O (6 mL), by heating in a microwave reactor at 90 °C for 2 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and analyzed by preparative HPLC (C0.05). 18 The solution was purified by column chromatography using an aqueous solution of MeCN + 0.1% HCO₂H₂O solution to give a yellow solid 5-chloro-N-(2,4-difluoro-3-(8-isopropyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (63.0 mg, 40% yield based on 55% purity). MS (ESI) m / z [M+H) + 568.25.

[0236] Step 4: N-(3-(2-amino-8-isopropyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-1) Using the universal method MO, 5-chloro-N-(2,4-difluoro-3-(8-isopropyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (63 mg, 0.061 mmol, 55%) and mCPBA (26 mg, 0.13 mmol) were reacted in CH2Cl2 (6 mL), followed by another portion of 3-chloroperoxybenzoic acid (8.2 mg, 0.043 mmol). After the reaction was complete, saturated NaHCO3 aqueous solution (5 mL) was added. After stirring, the layers were separated, the organic phase was concentrated under reduced pressure and dried under high vacuum to obtain an orange solid crude product, which was dissolved in i-PrOH (9 mL) and sonicated, and then NH4OH aqueous solution (28-30%, 0.32 mL, 2.4 mmol) was added dropwise over 30 minutes at room temperature. Another portion of an aqueous NH4OH solution (28-30%, 0.32 mL, 2.4 mmol) was added at room temperature, and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and analyzed by preparative HPLC (C60-2000 ppm).18 The solution was purified by filtration through a Waters PoraPak CX column, washed with MeOH, and eluted with 2 M NH3 in MeOH solution to give a white solid N-(3-(2-amino-8-isopropyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (8.0 mg, 24% yield based on 97% purity). MS (ESI) m / z [M+H) + 537.25. 1 H NMR (500 MHz, CD 3 OD ) δ ppm 8.53 (s, 1 H), 8.32 (d, J=2.32 Hz, 1 H), 8.02 - 8.07 (m, 1 H), 7.67 (s, 1 H), 7.47 (td, J=8.74, 5.87Hz, 1 H), 7.01 (t, J=8.80 Hz, 1 H), 5.86 (br. s., 1 H), 4.01 (s, 3 H), 1.56 (d, J=6.85 Hz, 6H).

[0237] Example 2: trans-5-chloro-N-(3-(2-(4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-2) Step 1: 6-Bromo-8-ethyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one A capped, dry microwave-safe vial containing 0.5 g (1.8 mmol) of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one and NaH (60% in mineral oil, 0.110 g, 2.8 mmol) was evacuated and purged with argon. Anhydrous DMF (10 mL) was added. After stirring at room temperature for 10 minutes, the reaction mixture was heated in an oil bath at 50 °C for 30 minutes, followed by a single addition of EtBr (0.21 mL, 2.8 mmol) at room temperature. After brief stirring at room temperature, the reaction mixture was stirred in an oil bath at 40 °C for 1.5 hours. After standing overnight at room temperature, the DMF was removed under reduced pressure, and the mixture was suspended in EtOH. H₂O was added to concentrate the mixture and remove the EtOH. The material was filtered, and the filter cake was washed with excess H2O to give a brownish-red solid, 6-bromo-8-ethyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (543.0 mg, 65% yield based on 66% purity). MS (ESI) m / z [M+H + 300.13 / 302.09.

[0238] Step 2: trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-ethylpyrido[2,3-d]pyrimidin-7(8H)-one Using the universal method MO, a solution of 6-bromo-8-ethyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.44 mmol, 66%) in CH2Cl2 (6 mL) was added at 0 °C to a solution of mCPBA (93 mg, 0.48 mmol) in CH2Cl2 (~1 mL). After 1.5 hours, the reaction was cooled to 0 °C again, and a slurry of mCPBA (45 mg, 0.26 mmol) in CH2Cl2 (1 mL) was added. The reaction was then heated to room temperature. After completion, a saturated aqueous solution of NaHCO3 (5 mL) was added. The layers were separated, and the organic layer was concentrated and dried under high vacuum. The solid residue was suspended in i-PrOH (9 mL) and sonicated. Add trans-N1,N1-dimethylcyclohexane-1,4-diamine*2HCl (284 mg, 1.3 mmol), followed by DIPEA (0.77 mL, 4.4 mmol) at room temperature. After sonication to remove solid lumps, the reaction mixture was stirred overnight at room temperature. Subsequently, the reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and analyzed by preparative HPLC (C6000-C6000 HPLC). 18The mixture was purified from an aqueous solution of MeCN + 0.1% HCO2H solution to obtain a pale yellow solid, trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-ethylpyridano[2,3-d]pyrimidin-7(8H)-one (198 mg, quantified). MS (ESI) m / z [M+H + 394.25 / 396.32.

[0239] Step 3: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-2) The solution was prepared using the general method SMC, with Cs2CO3 (147 mg, 0.452 mmol) in H2O (2 mL), 5-chloro-N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl)-2-methoxypyridine-3-sulfonamide (5.6 mL, 0.23 mmol, crude product in 0.04 M dioxane solution), 6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-ethylpyridano[2,3-d]pyrimidin-7(8H)-one (89 mg, 0.23 mmol) and PdCl2dppf•CH2Cl2 (18 mg, 0.023 mmol), by sealed heating at 90 °C for 1.5 h in a microwave reactor. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography using a solution of MeOH in CH2Cl2. The separated material was filtered through a Waters PoraPak CX column to give a white solid, trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (66 mg, 46% yield). 1H NMR (500 MHz, DMSO-d6) δ ppm 8.59 - 8.68 (m,1 H), 8.37 (d, J=2.45 Hz, 1 H), 8.03 (d, J=2.45 Hz, 1 H), 7.88 (d, J=7.46 Hz,1 H), 7.65 - 7.76 (m, 2 H), 7.19 (t, J=7.34 Hz, 1 H), 6.94 - 7.05 (m, 2 H), 4.23 - 4.34 (m, 2 H), 3.86 (s, 3 H), 3.71 (br. s., 1 H), 2.41 (s, 6H), 2.09 (d, J=9.90 Hz, 1 H), 1.81 - 2.01 (m, 3 H), 1.30 - 1.45 (m, 4H), 1.15 - 1.26 (m, 4H). MS (ESI)m / z [M+H] + 630.24.

[0240] Example 3: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-3) The solution was prepared using the general method SMC, with Cs2CO3 (147 mg, 0.452 mmol) in H2O (3 mL) and DME (5.4 mL), 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl)-2-methoxypyridine-3-sulfonamide (3.6 mL, 0.23 mmol, 0.063 M 1,4-dioxane solution), 6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-ethylpyridano[2,3-d]pyrimidin-7(8H)-one (89 mg, 0.23 mmol) and PdCl2dppf•CH2Cl2 (18 mg, 0.023 mmol), by heating in an oil bath at 110–90 °C for 2.8 h. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid SiO2 chromatography using a solution of MeOH in CH2Cl2. The collected material was then filtered through a Waters PoraPak CX column to give a beige solid of 5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (63 mg, 42% yield based on 95% purity). 1 H NMR (500 MHz, DMSO-d6) δ ppm 8.61 - 8.70 (m, 1 H), 8.44 - 8.52 (m, 1 H), 8.22 (s, 1 H), 8.16 (d,J=2.45 Hz, 1 H), 7.93 (d, J=7.34 Hz, 1 H), 7.70 - 7.79 (m, 1 H), 7.20 (d, J=4.40 Hz, 1 H), 7.12 - 7.17 (m, 1 H), 7.03 - 7.10 (m, 1 H), 4.23 - 4.38 (m, 2H), 3.96 (s, 3 H), 2.38 (d, J=11.37 Hz, 1 H), 2.32 (s,6H), 1.82 - 2.12 (m,4H), 1.30 - 1.43 (m,4H), 1.13 - 1.28 (m,4H). MS (ESI)m / z [M+H] + 630.24.

[0241] Example 4: trans-5-chloro-N-(3-(2-(4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-4) Step 1: (8-Methyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid The mixture was prepared using the universal method MB, with 6-bromo-8-methyl-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (100 mg, 0.35 mmol), B2pin2 (266 mg, 1.0 mmol), KOAc (120 mg, 1.2 mmol), and PdCl2dppf•CH2Cl2 (28 mg, 0.035 mmol) in anhydrous 1,4-dioxane (10 mL), by heating in an oil bath at 100 °C for 3 days and 20 hours. The crude mixture was used directly in the subsequent Suzuki-Miyaura coupling step. MS (ESI) m / z [M+H] + 252.13.

[0242] Step 2: 5-Chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide The solution was prepared using the general method SMC, with (8-methyl-2-(meththio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid (88 mg, 0.35 mmol, crude in 1,4-dioxane, 10 mL), Cs2CO3 (229 mg, 0.70 mmol), N-(3-bromo-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (145 mg, 0.35 mmol), PdCl2dppf•CH2Cl2 (28.7 mg, 0.035 mmol), and H2O (5 mL), by sealed heating in an oil bath at 90 °C–100 °C for 4 hours. The reaction mixture was then cooled to room temperature, and N-(3-bromo-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (116 mg, 0.28 mmol) and PdCl2dppf•CH2Cl2 (18 mg, 0.025 mmol) were added. After degassing, the mixture was heated in an oil bath at 90–100 °C for 1 day and 19 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO2, using CH2Cl2 solution of MeOH), followed by further purification by rapid chromatography (SiO2, using CH2Cl2 solution of EtOAc) to give a white solid 5-chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (29 mg, 14% yield based on 90% purity).

[0243] Step 3: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-4) To a suspension of 5-chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (29 mg, 0.054 mmol, 90%) in CH2Cl2 (10 mL), a solution of mCPBA (15.5 mg, 0.081 mmol) in CH2Cl2 (1 mL) was added at 0 °C. The mixture was stirred and slowly heated to room temperature for a total time of 1.9 hours. A saturated aqueous solution of NaHCO3 (5 mL) was then added. After stirring, the layers were separated. The organic layer was filtered through an anhydrous Na2SO4 short column, concentrated, and dried under high vacuum. The obtained crude product, trans-N1,N1-dimethylcyclohexane-1,4-diamine*2HCl (34.7 mg, 0.16 mmol), and DIPEA (0.094 mL, 0.54 mmol) were suspended in i-PrOH (8 mL) and sonicated. The suspension was stirred at room temperature for 6 days. The reaction mixture was then concentrated on diatomaceous earth and analyzed by preparative HPLC (C1000- ... 18 Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, the product yielded a white solid trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide carbamate (15.8 mg, 43% yield). 1 H NMR (500 MHz, DMSO-d6) δ 8.59-8.69 (m, 1H), 8.34 (s, 1H), 8.17 (br.s., 1H), 8.01 (d, J=1.59 Hz, 1H), 7.96 (d, J=7.34 Hz, 1H), 7.81 (d, , 2.64 (br. s., 1H), 2.45 (br.s., 6H), 1.84-2.16 (m, 4H), 1.30-1.50 (m, 4H). MS (ESI)m / z [M+H] + 634.23.

[0244] Example 5: N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-5) A 10 mL suspension of 5-chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (40 mg, 0.074 mmol)) in CH2Cl2 was added at 0 °C. The mixture was stirred under cooling and slowly heated to room temperature for a total of 2 hours. The reaction was then allowed to stand overnight at -20 °C. After reaching room temperature, 5 mL of saturated NaHCO3 aqueous solution was added. The organic layer was separated by stirring at room temperature, filtered through an anhydrous Na2SO4 short column, concentrated, and dried under vacuum. The residue was briefly sonicated in i-PrOH (8 mL), followed by the dropwise addition of 0.96 mL of NH4OH aqueous solution (7.4 mmol) at room temperature. The reaction was vigorously stirred at room temperature for 1.5 hours, then concentrated onto diatomaceous earth and analyzed by preparative HPLC (C10-C20). 18 Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, it yielded a white solid N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide*formic acid (10 mg, 24% yield). MS (ESI) m / z [M+H] + 509.18. 1 H NMR (500 MHz, DMSO-d6) δ 9.94-11.07 (br m, 1H), 8.57-8.70 (m, 1H), 8.40 (br. s., 1H), 8.03 (d, J=2.32 Hz, 1H), 7.75-7.86 (m,1H), 7.42 (s, 2H), 7.24 (d, J=5.75 Hz, 1H), 7.01 (br. s., 1H), 3.87 (s, 3H), 3.53 (s, 3H).

[0245] Example 6: 5-Chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-6) To a CH2Cl2 solution (10 mL) of 5-chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (42 mg, 0.078 mmol) cooled to -78 °C, a CH2Cl2 solution (2 mL) of mCPBA (20 mg, 0.082 mmol) was added. The mixture was stirred continuously and slowly heated to room temperature for a total time of 1.5 hours. A saturated aqueous solution of NaHCO3 was then added, and after brief stirring, the organic layer was separated, and the CH2Cl2 was concentrated to remove it. The recovered solid was then vigorously stirred at room temperature for 19.5 hours in i-PrOH (8 mL) and MeNH2 (2.0 M THF solution, 0.97 mL, 1.9 mmol) for 19.5 hours. The reaction mixture was then concentrated on diatomaceous earth and analyzed by preparative HPLC (C1000-2000 HPLC). 18 Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, it yielded a white solid 5-chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide*formic acid (10 mg, 23%). MS (ESI) m / z [M+H] + 523.25. 1 H NMR (500 MHz, DMSO-d6) δ 8.56-8.78 (m, 1H), 8.39 (br. s., 1H), 8.03 (d, J=1.96 Hz, 1H), 7.92 (d, J=3.30 Hz, 1H), 7.80 (s, 1H), 7.23 (d, J=5.87 Hz, 1H), 7.00 (br. s.,1H), 6.53 (br. s., 1H), 3.86 (s, 3H), 3.50-3.63 (m, 3H), 2.92 (d, J=4.40 Hz,3H).

[0246] Example 7: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-7) Step 1: trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one Solid trans-N1,N1-dimethylcyclohexane-1,4-diamine*2HCl (101 mg, 0.47 mmol), CsF (95 mg, 0.63 mmol), and 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (100 mg, 0.31 mmol) were added to DMSO (5 mL). DIPEA (0.55 mL, 3.1 mmol) was added, and the reaction mixture was heated at 50 °C for 2 days and 19 hours. The reaction was then partitioned between H₂O and EtOAc. The aqueous phase was extracted with EtOAc (2x). The combined organic layers were concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO2, using MeOH in CH2Cl2 solution 0 to 85%, followed by 2 M NH3-MeOH in CH2Cl2 solution) to give a pale orange membrane of trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpyridano[2,3-d]pyrimidin-7(8H)-one (55 mg, 45% yield based on 98% purity). MS (ESI) m / z [M+H] + 380.28 / 382.29.

[0247] Step 2: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans-I-7) The solution was prepared using the general method SMC, with 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl)-2-methoxypyridine-3-sulfonamide (2.3 mL, 0.14 mmol in 1,4-dioxane solution), trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one (55 mg, 0.14 mmol), Cs2CO3 (141 mg, 0.43 mmol), PdCl2dppf•CH2Cl2 (11.8 mg, 0.014 mmol), DME (5.6 mL), and H2O (4 mL), by sealed heating in an oil bath at 80 °C for 2.5 days. The reaction mixture was then concentrated under reduced pressure, deposited on diatomaceous earth, purified by rapid chromatography (SiO2, using a CH2Cl2 solution of MeOH), and then purified by preparative HPLC (C 18 Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, it yielded a white powder of trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide carboxylate (20 mg, yield 21% based on 98% purity). MS (ESI) m / z [M+H + 616.36. 1 H NMR (500 MHz, DMSO-d6) δ 8.59-8.69 (m,1H), 8.43 (d, J=2.20 Hz, 1H), 8.25 (br. s., 1H), 8.13 (d, J=2.45 Hz, 1H), 7.90 (d, J=7.58 Hz, 1H), 7.68-7.79 (m, 2H), 7.15 (d, J=4.40 Hz, 1H), 7.07-7.12 (m, 1H), 7.03 (br. s., 1H), 3.93 (s, 3H), 3.71-3.86 (m, 2H), 3.51-3.60(m, 3H), 2.23 (br. s., 6H), 1.80-2.11 (m, 4H), 1.26-1.37 (m, 4H).

[0248] Example 8: 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-8) Step 1: 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide 6-(5-amino-2-fluorophenyl)-8-methyl-2-(methylthio)pterin-7(8H)-one (200 mg, 0.62 mmol, 98%) and 5-chloro-2-methoxypyridine-3-sulfonyl chloride (164 mg, 0.68 mmol) were cooled at -10 °C, and then CH2Cl2 (20 mL) was added. After further cooling (10 min), pyridine (0.25 mL, 3.1 mmol) was added, and the vial was removed from the cooling bath, allowing the mixture to warm to room temperature with stirring. The reaction was stirred at room temperature for 4 days, then concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO2, using CH2Cl2 solution of EtOAc) to give a light orange solid, 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (259 mg, 80% yield). MS (ESI) m / z [M+H + 523.25.

[0249] Step 2: 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-8) To a CH2Cl2 solution (10 mL) of 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (60 mg, 0.11 mmol) cooled to -10 °C, a CH2Cl2 solution (2 mL) of mCPBA (30 mg, 0.12 mmol) was added. The reaction was heated to room temperature and stirred for a total of 18 hours at room temperature. CH2Cl2 was removed under reduced pressure. i-PrOH (8 mL) was added, followed by MeNH2 (40 wt% H2O solution, 0.50 mL, 5.7 mmol). After stirring at room temperature for 1.5 hours, the material was deposited on diatomaceous earth and analyzed by preparative HPLC (C60-C5 ... 18Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, it yielded a pale yellow solid, 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide carboxylic acid (25 mg, yield 39% based on 99% purity). MS (ESI) m / z [M+H + 506.25. 1 H NMR (500MHz, DMSO-d6) δ 10.64 (br. s., 1H), 8.63-8.78 (m, 1H), 8.47 (d, J=2.45 Hz, 1H), 7.93-8.23 (m, 2H), 7.30 (d, J=5.87 Hz, 1H), 7.18 (d, J=7.21 Hz, 2H), 3.95 (s, 3H), 3.46-3.61 (m, 3H), 2.93 (d, J=4.65 Hz, 3H).

[0250] Example 9: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-9) To a CH₂Cl₂ (10 mL) solution of 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (70 mg, 0.134 mmol) cooled to -78 °C, a CH₂Cl₂ (2 mL) solution of mCPBA (35 mg, 0.14 mmol) was added. The reaction was then slowly heated to room temperature and stirred at room temperature for a total time of 18 hours. CH₂Cl₂ was removed under reduced pressure. trans-N,N,1-dimethylcyclohexane-1,4-diamine*2HCl (89 mg, 0.41 mmol) was added. The entire mixture was suspended in i-PrOH (8 mL), sonicated, and then DIPEA (0.23 mL, 1.3 mmol) was added. The mixture was vigorously stirred at room temperature for 2.5 hours. Then, more i-PrOH (8 mL) was added, followed by brief sonication and stirring overnight at room temperature. The reaction was then heated in an oil bath at 50 °C for 3 hours. Subsequently, the reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and analyzed by preparative HPLC (C10-C20). 18Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, it yielded a pale yellow solid, 5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (16.5 mg, yield 18% based on 98% purity). MS (ESI) m / z [M+H + 617.30. 1 H NMR (500 MHz, DMSO-d6) δ 8.55-8.69 (m,1H), 8.38 (d, J=2.57 Hz, 1H), 8.16 (br. s., 1H), 7.87-8.07 (m, 2H), 7.20 (d,J=5.38 Hz, 1H), 7.09 (d, J=7.70 Hz, 2H), 3.87 (s, 3H), 3.64-3.82 (m, 1H), 3.42-3.47 (m, 3H), 2.27-2.37 (m, 1H), 2.22 (s, 6H), 1.76-2.04 (m, 4H), 1.21-1.38 (m, 4H).

[0251] Example 10: trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (trans I-10) The sample was prepared using the general method SMC, with PdCl2dppf•CH2Cl2 (15.0 mg, 0.018 mmol), Cs2CO3 (180 mg, 0.55 mmol), trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (70 mg, 0.18 mmol), H2O (1.8 mL), and (3-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (3.5 mL, 0.18 mmol, 0.052 M crude 1,4-dioxane solution), by heating in a microwave reactor at 90 °C for 100 min. The sample was then analyzed by rapid chromatography (SiO2, using a CH2Cl2 solution of MeOH) followed by preparative HPLC (C 18Purified using an aqueous solution of MeCN + 0.1% HCO2H solution, it yielded a white solid trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide carboxylate (23 mg, yield 17% based on 99% purity). MS (ESI) m / z [M+H + 616.43. 1 H NMR (500 MHz, DMSO-d6) δ 8.57-8.68 (m,1H), 8.33-8.40 (m, 1H), 8.18 (d, J=10.39 Hz, 1H), 8.03 (d, J=2.57 Hz, 1H), 7.82-7.91 (m, 1H), 7.67-7.75 (m, 1H), 7.19 (t, J=7.64 Hz, 1H), 6.98-7.03 (m,1H), 6.94 (br. s., 1H), 3.82-3.90 (m, 3H), 3.75 (br. s., 1H), 3.50-3.58 (m,3H), 2.41 (br. s., 6H), 1.85-2.14 (m, 4H), 1.27-1.49 (m, 4H).

[0252] Example 11: trans-5-chloro-N-(2-cyano-3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (trans I-11) Step 1: N-(3-bromo-2-cyanophenyl)-5-chloro-N-((5-chloro-2-methoxypyridin-3-yl)sulfonyl)-2-methoxypyridin-3-sulfonamide Solid 5-chloro-2-methoxypyridine-3-sulfonyl chloride (307 mg, 1.27 mmol) and 2-amino-6-bromobenzonitrile (250 mg, 1.27 mmol) were cooled at -78 °C, and then CH2Cl2 (10 mL) and pyridine (0.51 mL, 6.3 mmol) were added. The reaction mixture was then removed from the cooling bath and stirred at room temperature for 2 days. Subsequently, another portion of 5-chloro-2-methoxypyridine-3-sulfonyl chloride (307 mg, 1.27 mmol) was added at room temperature, and stirring was continued for 3 days. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO2, using a hexane solution of EtOAc) to give a white powder, N-(3-bromo-2-cyanophenyl)-5-chloro-N-((5-chloro-2-methoxypyridin-3-yl)sulfonyl)-2-methoxypyridin-3-sulfonamide (333 mg, 43%). MS (ESI) m / z [M+H] + 606.99 / 609.1.

[0253] Step 2: (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-cyanophenyl)boronic acid Preparation using the general method MB: N-(3-bromo-2-cyanophenyl)-5-chloro-N-((5-chloro-2-methoxypyridin-3-yl)sulfonyl)-2-methoxypyridin-3-sulfonamide (100 mg, 0.16 mmol), B2pin2 (63 mg, 0.25 mmol), KOAc (48 mg, 0.49 mmol), and PdCl2dppf•CH2Cl2 (13 mg, 0.016 mmol) in 1,4-dioxane (5 mL) was microwave-controlled at 100 °C for 2 h, followed by heating in an oil bath at 100 °C for 1 day and 19 h. The entire crude product was used for the next step. MS ESI [M+H] + 368.18.

[0254] Step 3: trans-5-chloro-N-(2-cyano-3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (trans I-11) SMC was prepared using a general method: (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-cyanophenyl)boronic acid (4.9 mL, 0.16 mmol, 5.0 mL, 0.033 M crude 1,4-dioxane solution), PdCl2dppf•CH2Cl2 (13 mg, 0.016 mmol), Cs2CO3 (266 mg, 0.81 mmol), trans-6-bromo-2-((4-(dimethylamino)cyclohexyl)amino)-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (62 mg, 0.16 mmol) and H2O (2.5 mL) were used in a microwave reactor under sealed heating at 90 °C for 2 hours. The solution was purified by rapid chromatography (using a CH2Cl2 solution of MeOH) and by preparative HPLC (C 18 The mixture was repurified using an aqueous solution of MeCN + 0.1% HCO2H solution to give a white solid trans-5-chloro-N-(2-cyano-3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (38 mg, 33%). MS (ESI) m / z [M+H] + 623.43. 1 H NMR (500 MHz, DMSO-d6) δ 10.83 (br. s., 0.3H), 9.33 (d, J=8.44Hz, 0.6H), 8.64-8.75 (m, 1H), 8.39 (br. s., 1H), 8.12-8.19 (m, 1H), 7.99-8.07(m,1H), 7.92 (d, J=2.69 Hz, 1H), 7.78-7.90 (m, 1H), 7.47 (br. s., 1H), 7.25(d, J=8.07 Hz, 1H), 3.84-3.86 (m, 4H), 3.53-3.63 (m, 3H), 3.09-3.25 (m, 1H), 2.72-2.79 (m, 6H), 1.98-2.20 (m, 4H), 1.31-1.69 (m, 4H).

[0255] Example 12: 5-Chloro-N-(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-12) Step 1: 6-Bromo-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one Prepared using general method NS, with a suspension of 6-bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one (600 mg, 2.082 mmol) and MeNH2 (40 wt% H2O solution, 3.6 mL, 42 mmol) in i-PrOH (20 mL). The reaction was shaken at room temperature for 10 min, then stirred at room temperature for 19 h. Subsequently, the reaction was shaken again at room temperature for 2 h. Filtered and washed with EtOH, yielding a white solid of 6-bromo-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (414 mg, 78% yield). MS (ESI) m / z [M+H] + 255.10|257.20.

[0256] Step 2: 5-Chloro-N-(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-12) The mixture was prepared using the general method SMC with Cs₂CO₃ (644 mg, 1.98 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (261 mg, 0.72 mmol), 6-bromo-2-(methylamino)pyridino[2,3-d]pyrimidin-7(8H)-one (168 mg, 0.66 mmol), and PdCl₂dppf•CH₂Cl₂ (54 mg, 0.066 mmol) in H₂O (5 mL) and 1,4-dioxane (10 mL); by sealed heating in an oil bath at 95 °C for 22 h. The crude mixture was then cooled to room temperature and partitioned between H₂O and CH₂Cl₂. The aqueous layer was extracted with a 2% v / v MeOH solution in CH₂Cl₂ (2x). The combined organic layers were concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography to give a white solid, 5-chloro-N-(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (62.0 mg, 19% yield). MS (ESI) m / z [M+H + 491.27. 1H NMR (500 MHz, DMSO-d6)δ ppm 11.82 - 12.11 (m, 1 H), 10.37 (br. s., 1 H), 8.53 - 8.70 (m, 1 H), 8.46(d, J=2.32 Hz, 1 H), 8.05 (d, J=2.45 Hz,1 H), 7.65 - 7.81 (m, 1.6 H), 7.45 -7.57 (m, 0.4 H), 7.25 (dt, J=14.09, 6.95 Hz, 2 H), 7.12 - 7.19 (m, 1 H), 3.89(s, 3 H), 2.86 (d,J=4.65 Hz, 3H).

[0257] Example 13: 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-13) To a CH2Cl2 solution (20 mL) of 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (129 mg, 0.25 mmol) cooled to 0 °C, a CH2Cl2 solution (1-2 mL) of mCPBA (64 mg, 0.26 mmol) was added, and the mixture was stirred and slowly heated to room temperature for a total of 4 hours. 50% of the crude mixture was concentrated to remove CH2Cl2. The solid residue was suspended in i-PrOH (6 mL) and treated with i-PrNH2 (0.32 mL, 3.7 mmol) at room temperature. The suspension was stirred at room temperature for 1 day and 19 hours, then concentrated onto diatomaceous earth and analyzed by preparative HPLC (C10-C2 ... 18 Purification with an aqueous solution of MeCN + 0.1% HCO2H solution yielded a white solid, 5-chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (28 mg, 40% yield based on 94% purity). MS (ESI) m / z [M+H + 534.53. 1HNMR (500 MHz, DMSO-d6) δ ppm 8.62 - 8.75 (m, 1 H), 8.46 (d, J=2.32 Hz, 1 H), 8.13 (d, J=2.45 Hz, 1 H), 7.88 - 8.08 (m, 1 H), 7.29 (d, J=5.75 Hz, 1 H), 7.12 - 7.22 (m, 2 H), 4.09 - 4.26 (m, 1 H), 3.95 (s, 3 H), 1.15 - 1.27 (m, 6H). The N-Me signal is assumed to be masked by the "H2O" peak.

[0258] Example 14: 5-Chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-14) Prepared using the same method as for 5-chloro-N-(3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide, with EtNH2 (66-72% H2O solution), to obtain a pale yellow solid of 5-chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (25.0 mg, 40% yield). MS (ESI) m / z [MH - 518.31. 1 ¹H NMR (500MHz, DMSO-d⁶) δ ppm 10.55 (br. s., 1 H), 8.61 - 8.78 (m, 1 H), 8.47 (d, J=2.45 Hz, 1 H), 8.14 (d, J=2.57 Hz, 2 H), 7.96 - 8.06 (m, 1 H), 7.30 (d, J=5.99 Hz, 1 H), 7.11 - 7.24 (m, 2 H), 3.96 (s, 3 H), 1.06 - 1.29 (m, 3 H). The N-Me signal is assumed to be masked by the “H₂O” peak.

[0259] Example 15: 5-Chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-15) Step 1: 6-(3-amino-2,6-difluorophenyl)-8-methyl-2-(methylamino)pterin-7(8H)-one A mixture of 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (153 mg, 0.60 mmol), 6-bromo-8-methyl-2-(methylamino)pterin-7(8H)-one (67.5 mg, 0.25 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (29.5 mg, 0.037 mmol), and K3PO4 (186 mg, 0.875 mmol) was suspended in 1,4-dioxane (5 mL), and then H2O (0.5 mL) was added. The vial was sealed and degassed. The mixture was then heated in a microwave reactor at 65 °C for 5 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (using MeOH in CH2Cl2 solution) to give a pale yellow solid, 6-(3-amino-2,6-difluorophenyl)-8-methyl-2-(methylamino)pterin-7(8H)-one (67 mg, 84% yield). MS (ESI) m / z [M+H] + 319.41.

[0260] Step 2: 5-Chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-15) Prepared using a general method (NS): using CH2Cl2 (11 mL), 6-(3-amino-2,6-difluorophenyl)-8-methyl-2-(methylamino)pterin-7(8H)-one (67 mg, 0.211 mmol), 5-chloro-2-methoxypyridine-3-sulfonyl chloride (56 mg, 0.23 mmol), and pyridine (0.093 mL, 1.2 mmol). After completion, the reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, purified by rapid chromatography (using a CH2Cl2 solution of MeOH), and then by preparative HPLC (C60-2000 ppm). 18Purification with an aqueous solution of MeCN + 0.1% HCO2H solution yielded a white solid (slightly pale yellow tinge) of 5-chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (43 mg, yield 39% based on 97% purity). MS (ESI) m / z [M+H) + 524.39. 1 ¹H NMR (500 MHz, DMSO-d⁶) δ ppm 8.76 (s, 0.3 H), 8.68 (s, 0.7 H), 8.43–8.50 (m, 1 H), 8.16 (d, J=4.77 Hz, 1 H), 8.05 (d, J=2.57 Hz, 1 H), 7.34–7.45 (m, 1 H), 7.15 (t, J=8.80 Hz, 1 H), 3.89 (s, 3 H), 2.88–2.99 (m, 3 H). An N-Me signal is hidden beneath the solvent peak.

[0261] Example 16: 5-Chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (I-16) Step 1: 6-Bromo-2-(ethylamino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one A mixture of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (300 mg, 0.94 mmol), i-PrOH (10 mL), and EtNH2 (66-72% H2O solution, 1.5 mL, 19 mmol) was stirred at room temperature for 1 day. The solid was collected by filtration and washed with EtOH to give a white solid of 6-bromo-2-(ethylamino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one (227 mg, 85% yield). MS (ESI) m / z [M+H] + 282.94|284.97.

[0262] Step 2: 6-(3-amino-2,6-difluorophenyl)-2-(ethylamino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one Preparation using the general method of SMC: 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (130 mg, 0.51 mmol), 6-bromo-2-(ethylamino)-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (60 mg, 0.21 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (25 mg, 0.032 mmol), and K3PO4 (157 mg, 0.74 mmol) in 1,4-dioxane (5 mL) and H2O (0.5 mL) in a microwave reactor under sealed heating at 65 °C for 10 h. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (using MeOH in CH2Cl2 solution) to give a light brown solid, 6-(3-amino-2,6-difluorophenyl)-2-(ethylamino)-8-methylpyridano[2,3-d]pyrimidin-7(8H)-one (83 mg, quantitative yield). MS (ESI) m / z [M+H] + 332.20.

[0263] Step 3: 5-Chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (I-16) The reaction mixture was prepared using the general method NS with CH2Cl2 (11 mL), 6-(3-amino-2,6-difluorophenyl)-2-(ethylamino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one (70 mg, 0.21 mmol), 5-chloro-2-methoxypyridine-3-sulfonyl chloride (56 mg, 0.23 mmol), and pyridine (0.094 mL, 1.2 mmol) at 0 °C. After stirring at room temperature for 1 day and 18 hours, the reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and analyzed by preparative HPLC (C60-C5 ... 18 The mixture was purified by aqueous solution of MeCN + 0.1% HCO2H solution, and then re-purified twice by rapid chromatography (using CH2Cl2 solution with CH2Cl2 / MeOH / concentrated NH4OH at a ratio of 89 / 10 / 1) to give a white solid 5-chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (9.0 mg, 8% yield). MS (ESI) m / z [M+H) + 537.42.1 H NMR (500 MHz, CD 3 OD ) δ ppm 8.40 - 8.50 (m, 1 H), 8.16- 8.23 ​​(m, 1 H), 7.95 (d, J=2.57 Hz, 1 H), 7.62 (s, 1 H), 7.37 (td, J=8.86,5.75 Hz, 1 H), 6.91 (td, J=8.89, 1.41 Hz, 1 H), 3.86 - 3.92 (m, 3 H), 3.51 -3.66 (m, 3 H), 3.43 (d, J=6.72 Hz, 2 H), 1.10 - 1.24 (m, 3 H).

[0264] Example 17: 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-17) Step 1: 6-Bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one A cold (0 °C) suspension of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (500 mg, 1.8 mmol) and mCPBA (476 mg, 1.9 mmol) in 100 mL CH₂Cl₂ was slowly heated to room temperature with stirring for a total time of 1 day and 19 hours. The product was separated by filtration and washed with CH₂Cl₂ to give a white solid of 6-bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one (490 mg, 93%). MS (ESI) m / z [M+H] + 288.24 |290.25.

[0265] Step 2: 6-Bromo-2-(ethylamino)pyrido[2,3-d]pyrimidin-7(8H)-one A single addition of EtNH2 (66-72% H2O solution, 0.55 mL, 6.9 mmol) to an i-PrOH suspension of 100 mg (100 mg, 0.35 mmol) of 6-bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one was made. After stirring at room temperature for 21 hours, the product was collected by filtration and washed with EtOH to give a white solid of 6-bromo-2-(ethylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (70 mg, 75% yield). MS (ESI) m / z [M+H] + 269.25 |271.25.

[0266] Step 3: 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-17) SMC was prepared using a general method: 6-bromo-2-(ethylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (60 mg, 0.22 mmol), PdCl2dppf•CH2Cl2 (21.24 mg, 0.026 mmol), Cs2CO3 (212 mg, 0.650 mmol), (5-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (113 mg, 0.31 mmol), H2O (2 mL), and 1,4-dioxane (4 mL) were used in a microwave reactor and heated in a sealed container at 100 °C for 2.5 h. The mixture was purified by rapid chromatography (using a CH2Cl2 solution of MeOH) and then by preparative HPLC (C 18 Purification with an aqueous solution of MeCN + 0.1% HCO₂H₂O solution yielded a white solid 5-chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (20.0 mg, 18% yield). MS (ESI) m / z [M+H] + 505.33. 1¹H NMR (500 MHz, DMSO-d⁶) δ ppm 11.97 (br. s., 1 H), 8.54 - 8.68 (m, 1 H), 8.44 (d, J=1.59 Hz, 1 H), 8.13 (d, J=1.83 Hz, 1 H), 7.76 (br. s., 0.5 H), 7.70 (s, 1 H), 7.58 (br. s., 0.5 H), 7.19 (d, J=4.28 Hz, 1 H), 7.09 -7.16 (m, 1 H), 7.07 (br. s., 1 H), 3.95 (s, 3 H), 1.03 - 1.25 (m, 3 H). (A CH₂ signal is masked by the "solvent / H₂O" peak).

[0267] Example 18: 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-18) Step 1: 6-Bromo-2-(isopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one i-PrNH2 (0.60 mL, 6.9 mmol) was added in a single batch to a suspension of 10 mL of i-PrOH containing 100 mg (100 mg, 0.35 mmol) of 6-bromo-2-(isopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one. After stirring at room temperature for 3 days, the product was collected by filtration and washed with EtOH to give a white solid of 6-bromo-2-(isopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (84.0 mg, 85% yield). MS (ESI) m / z [M+H] + 283.24|285.27.

[0268] Step 2: 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-18) SMC was prepared using a general method: 6-bromo-2-(isopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (73 mg, 0.26 mmol), PdCl2dppf•CH2Cl2 (21 mg, 0.026 mmol), Cs2CO3 (210 mg, 0.64 mmol), and (5-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (112 mg, 0.31 mmol), H2O (2 mL), and 1,4-dioxane (4 mL) were used in a microwave reactor and heated in a sealed container at 100 °C for 2.5 h. The mixture was purified by rapid chromatography (using a CH2Cl2 solution of MeOH) and then by preparative HPLC (C 18 The mixture was purified with an aqueous solution of MeCN and 0.1% HCO₂H₂ solution, followed by grinding with MeCN to give a white solid 5-chloro-N-(4-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (12.0 mg, 9% yield based on 95% purity). MS (ESI) m / z [M+H) + 519.31. 1 H NMR (500 MHz, DMSO-d6) δ ppm11.94 (br. s., 1 H), 8.52 - 8.69 (m, 1 H), 8.43 (d, J=1.83 Hz, 1 H), 8.12 (d,J=1.96 Hz, 1 H), 7.60 - 7.76 (m, 1.7 H), 7.41 - 7.53 (m, 0.3 H), 7.18 (d, J=4.16 Hz, 1 H), 7.08 - 7.15 (m, 1 H), 7.07 (br. s., 1 H), 4.14 (br. s., 1 H), 3.95 (s, 3 H), 1.17 (d, J=6.24 Hz, 6H).

[0269] Example 19: 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-19) Step 1: 6-Bromo-2-(isopropylamino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one i-PrNH2 (2.43 mL, 28 mmol) was added dropwise to an i-PrOH (10 mL) suspension of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (300 mg, 0.94 mmol). After stirring at room temperature for 1 day and 3 hours, the product was collected by filtration and washed with EtOH to give a white solid of 6-bromo-2-(isopropylamino)-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one (110.0 mg, 39% yield). MS (ESI) m / z [M+H] + 296.93 / 298.96.

[0270] Step 2: 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-19) SMC was prepared using a general method: 6-bromo-2-(isopropylamino)-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (77 mg, 0.26 mmol), PdCl2dppf•CH2Cl2 (21 mg, 0.026 mmol), Cs2CO3 (211 mg, 0.65 mmol), and (5-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (112 mg, 0.31 mmol) were heated in a microwave reactor at 90 °C for 1.5 h in H2O (2 mL) and 1,4-dioxane (4 mL). The mixture was purified by rapid chromatography (using a CH2Cl2 solution of MeOH) and then by preparative HPLC (C 18 The mixture was purified by an aqueous solution of MeCN plus 0.1% HCO2H solution to give a white powdery 5-chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (73.0 mg, 53% yield). 1¹H NMR (500 MHz, DMSO-d⁶) δ ppm 8.58 - 8.70 (m, 1 H), 8.45 (d, J=2.45 Hz, 1 H), 8.14 (d, J=2.57 Hz, 1 H), 7.83 (d, J=7.34 Hz, 0.5 H), 7.64 -7.76 (m, 1.5 H), 7.20 (dd, J=6.36, 2.69 Hz, 1 H), 7.11 - 7.17 (m, 1 H), 7.05 - 7.10 (m, 1 H), 4.09 - 4.30 (m, 1 H), 3.86 - 3.98 (m, 3 H), 1.07 - 1.44 (m, 6 H). (CH₃ signal is masked). MS (ESI)m / z [M+H] + 533.48.

[0271] Example 20: 5-Chloro-N-(2-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-20) SMC was prepared using a general method: 6-bromo-8-methyl-2-(methylamino)pterin-7(8H)-one (67.5 mg, 0.25 mmol), Cs₂CO₃ (293 mg, 0.90 mmol), PdCl₂dppf•CH₂Cl₂ (20 mg, 0.025 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (108 mg, 0.30 mmol), 1,4-dioxane (4.8 mL), and H₂O (2.4 mL) were heated in a microwave reactor at 90 °C for 1.5 hours in a sealed container. Purification by rapid chromatography (using CH2Cl2 / MeOH / concentrated NH4OH in a CH2Cl2 solution of 89 / 10 / 1) yielded a pale yellow solid, 5-chloro-N-(2-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (18.0 mg, 14% yield). MS (ESI) m / z [M+H] + 506.41. 1¹H NMR (500 MHz, DMSO-d⁶) δ ppm: 10.43 (br. s., 1 H), 8.73 (s, 0.3 H), 8.65 (s, 0.7 H), 8.46 (d, J=2.45 Hz, 1 H), 8.01 - 8.09 (m, 1.7 H), 7.93 (d, J=4.28 Hz, 0.3 H), 7.29 - 7.41 (m, 2 H), 7.14 - 7.27 (m, 1 H), 3.89 (s, 3 H), 2.93 (d, J=4.77 Hz, 3 H). A “Me” signal was masked by the solvent peak.

[0272] Example 21: 5-Chloro-N-(3-(2-(cyclopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-21) Solid mCPBA (173 mg, 0.70 mmol) was added to a stirred, cold (0 °C) suspension of 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (350 mg, 0.67 mmol) in CH2Cl2 (20 mL), and the mCPBA was transferred using CH2Cl2 (1 mL). The reaction was then stirred, and the mixture was slowly heated to room temperature and then stirred overnight at room temperature. At this point, the conversion is complete, and the reaction mixture contains 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylsulfinyl)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide and 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylsulfinyl)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide. The reaction mixture is aliquoted into six equal portions, each concentrated under reduced pressure, and used directly for subsequent S-phase reactions without further purification. N Ar steps. MS (ESI)m / z [M+H] + 539.30 and 555.29.

[0273] One-sixth fraction of the solids from the mCPBA oxidation step was suspended in i-PrOH (10 mL) and treated with c-PrNH2 (0.23 mL, 3.3 mmol), followed by shaking at room temperature for 3 days and 22 hours. The solids were collected by filtration and washed with EtOH to give a pale yellow solid of 5-chloro-N-(3-(2-(cyclopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (49.3, 83% yield). MS (ESI) m / z [M+H] + 532.28. 1 ¹H NMR (500 MHz, DMSO-d⁶) δ ppm 9.99 - 10.94 (m, 1 H), 8.62 - 8.80 (m, 1 H), 8.41 - 8.50 (m, 1 H), 8.17 - 8.33 (m, 1 H), 8.02 - 8.15 (m, 1 H), 7.29 (d, J=6.36 Hz, 1 H), 7.12 - 7.22 (m, 2 H), 3.95 (s, 3 H), 2.89 (br. s., 1 H), 0.76 (br. s., 2 H), 0.56 (br. s., 2 H) correspond to N-Me signals that are masked by solvent peaks.

[0274] Example 22: (R)-5-chloro-N-(4-fluoro-3-(8-methyl-7-oxo-2-((tetrahydrofuran-3-yl)amino)-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide ((R)-I-22) Similar to the synthesis of 5-chloro-N-(3-(2-(cyclopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide, a 1 / 6 portion of the mCPBA oxidation product from 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (350 mg, 0.669 mmol) was suspended in i-PrOH (10 mL) and treated with (R)-3-aminotetrahydrofuran (0.29 mL, 3.3 mmol) and shaken at room temperature for 3 days and 22 hours. The solid was collected by filtration and washed with EtOH to give a pale yellow solid (R)-5-chloro-N-(4-fluoro-3-(8-methyl-7-oxo-2-((tetrahydrofuran-3-yl)amino)-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (46 mg, 73% yield based on 99% purity). MS (ESI) m / z [M+H + 562.37. 1 H NMR (500 MHz, DMSO-d6) δ ppm 8.66 - 8.75 (m, 1 H), 8.44 (d, J=2.57 Hz, 1 H), 8.39 (d, J=5.87 Hz, 0.7 H), 8.25 (d, J=5.01 Hz, 0.3 H), 8.12(d, J=2.57 Hz, 1 H), 7.28 (d, J=5.99 Hz, 1 H), 7.17 (d, J=7.58 Hz, 2 H), 4.43- 4.56 (m, 2 H), 3.97 - 4.01 (m, 1 H), 3.95 (s, 3 H), 3.82 - 3.91 (m, 2 H),3.71 - 3.79 (m, 2 H), 2.16 - 2.29 (m, 1 H), 1.88 - 2.03 (m, 1 H).

[0275] Example 23: 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(oxetane-3-ylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-23) Similar to the synthesis of 5-chloro-N-(3-(2-(cyclopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide, a 1 / 6 portion of the mCPBA oxidation product from 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (350 mg, 0.669 mmol) was suspended in i-PrOH (10 mL) and treated with 3-oxacyclobutylamine (269 mg, 3.7 mmol). The reaction mixture was shaken at room temperature for 3 days and 22 hours. The solid was collected by filtration and washed with EtOH to give a pale yellow solid, 5-chloro-N-(4-fluoro-3-(8-methyl-2-(oxetane-3-ylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (30.3 mg, 47% yield based on 95% purity). MS (ESI) m / z [M+H + 548.38. 1 ¹H NMR (500 MHz, DMSO-d⁶) δ ppm 10.27 (br.s, 1 H), 8.73 - 8.85 (m, 1 H), 8.71 (s, 1 H), 8.45 (d, J=2.57 Hz, 1 H), 8.13 (d, J=2.57 Hz, 1 H), 7.29 (d, J=6.24 Hz, 1 H), 7.09 - 7.23 (m, 2 H), 4.94 - 5.12 (m, 1 H), 4.73 - 4.88 (m, 2 H), 4.46 - 4.66 (m, 2 H), 3.95 (s, 3 H). The N-Me signal was masked by the solvent peak.

[0276] Example 24: 5-Chloro-N-(4-fluoro-3-(8-methyl-7-oxo-2-((tetrahydro-2H-pyran-4-yl)amino)-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-24) Similar to the synthesis of 5-chloro-N-(3-(2-(cyclopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide, a 1 / 6 portion of the mCPBA oxidation product from 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylthio)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (350 mg, 0.67 mmol) was suspended in i-PrOH (10 mL) and treated with tetrahydropyran-4-ylamine (113 mg, 1.1 mmol). The reaction mixture was shaken at room temperature for 3 days and 22 hours. The solid was collected by filtration and washed with EtOH to give a pale yellow solid of 5-chloro-N-(4-fluoro-3-(8-methyl-7-oxo-2-((tetrahydro-2H-pyran-4-yl)amino)-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (39.4 mg, 61% yield based on 99% purity). MS (ESI) m / z [MH] - 574.34. 1 H NMR (500 MHz, DMSO-d6) δ ppm 9.79 - 10.91 (m, 1 H),8.62 - 8.77 (m, 1 H), 8.46 (d, J=2.45 Hz, 1 H), 8.17 (d, J=7.34 Hz, 0.7 H),8.13 (d,J=2.57 Hz, 1 H), 8.05 (d, J=7.83 Hz, 0.3 H), 7.29 (d, J=5.6 Hz, 1H), 7.18 (d, J=7.58 Hz, 2 H), 3.99 - 4.18 (m, 1 H), 3.95 (s, 3 H), 3.83 - 3.93(m, 3 H), 1.76 - 1.97 (m, 2 H), 1.43 - 1.66 (m, 2 H). The CH and N-Me signals are masked by the solvent peaks.

[0277] Example 25: 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-25) Step 1: 6-Bromo-2-(ethylamino)pyrido[2,3-d]pyrimidin-7(8H)-one Solid mCPBA (761 mg, 3.1 mmol) was added to a cold (0 °C) suspension of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (800 mg, 2.9 mmol) in CH₂Cl₂ (200 mL). The reaction mixture was then stirred and allowed to slowly heat to room temperature for a total time of 1 day and 19 hours. After washing with CH₂Cl₂, the material was collected by filtration to give a white solid of 6-bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one (759.0 mg, 86% yield based on 96% purity). MS (ESI) m / z [M+H] + 288.13|290.16.

[0278] 6-Bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one (159 mg, 0.22 mmol) was suspended in i-PrOH (10 mL), and EtNH2 (66-72% H2O solution, 0.93 mL, 11 mmol) was added in a single addition at room temperature. The reaction mixture was stirred at room temperature for 19 hours. The precipitate was collected by filtration and washed with EtOH to give 6-bromo-2-(ethylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (128 mg, 86% yield) as a white solid. MS (ESI) m / z [M+H] + 269.17 | 271.20.

[0279] Step 2: 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-25) Preparation was performed using the general method of SMC: Cs₂CO₃ (465 mg, 1.427 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (171 mg, 0.48 mmol), 6-bromo-2-(ethylamino)pyridino[2,3-d]pyrimidin-7(8H)-one (128 mg, 0.48 mmol), and PdCl₂dppf•CH₂Cl₂ (39 mg, 0.048 mmol) were heated in an oil bath at 95 °C for 23 h in H₂O (5 mL) and 1,4-dioxane (10 mL). The reaction mixture was cooled to room temperature and partitioned between H₂O and CH₂Cl₂. The aqueous layer was extracted with CH₂Cl₂ containing a small amount of MeOH (2x). The combined organic layers were concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (using MeOH|CH2Cl2|concentrated NH4OH 89:10:1) to give a white solid 5-chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (60.0 mg, 25% yield). MS (ESI) m / z [M+H] + 505.29. 1 H NMR (500 MHz, DMSO-d6) δ ppm 1.07 - 1.17 (m, 3 H)3.35 (br d, J =5.50 Hz, 2 H)3.88(s, 3 H)7.11 - 7.18 (m, 1 H)7.19 - 7.29 (m, 2 H)7.51 - 7.63 (m, 0.3 H)7.69(s, 1 H)7.73 - 7.85 (m, 1 H)7.98 - 8.15 (m, 0.7 H)8.46 (d, J =2.45 Hz, 1 H)8.51 - 8.70 (m, 1 H)10.25 - 10.47 (m, 1 H)11.82 -12.05 (m, 1 H).

[0280] Example 26: 5-Chloro-N-(4-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-26) Preparation was performed using the general method SMC: 6-bromo-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (120 mg, 0.47 mmol), PdCl2dppf•CH2Cl2 (38 mg, 0.047 mmol), Cs2CO3 (383 mg, 1.2 mmol), and (5-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (204 mg, 0.56 mmol) were heated in a microwave reactor at 100 °C for 4 hours in H2O (5 mL) and 1,4-dioxane (10 mL). The reaction mixture was partitioned between H2O and CH2Cl2. The aqueous layer was extracted with CH2Cl2 containing a small amount of MeOH (2x). The combined organic layers were concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (using MeOH|CH2Cl2|concentrated NH4OH 89:10:1) to give a white solid 5-chloro-N-(4-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (129.0 mg, 56% yield). MS (ESI) m / z [M+H + 491.35. 1 H NMR (500MHz, DMSO-d6) δ ppm 11.78 - 12.17 (m, 1 H), 10.23 - 10.72 (m, 1 H), 8.53 -8.68 (m, 1 H), 8.45 (d, J= 2.4 Hz), 8.14 (d, J=2.4 Hz, 1 H), 7.66 - 7.75 (m,1.7 H), 7.50 (br s, 0.3 H), 7.20 (dd, J =6.24, 2.32 Hz, 1 H), 7.11 - 7.16 (m, 1 H), 7.05 - 7.10 (m, 1 H), 3.96 (s, 3 H), 2.83 - 2.88 (m, 3 H).

[0281] Example 27: 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-27) Step 1: 6-Bromo-8-methyl-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one To a suspension of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (500 mg, 1.57 mmol) in i-PrOH (17 mL), MeNH2 (40 wt% H2O solution, 3.4 mL, 39 mmol) was added at room temperature, and the reaction mixture was shaken at room temperature for 6 days and 23 hours. The precipitate was collected by filtration and washed with i-PrOH to give a white solid of 6-bromo-8-methyl-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (345.0 mg, 82% yield). MS (ESI) m / z [M+H] + 269.24|271.20.

[0282] Step 2: 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-27) Preparation using the general method SMC: 6-bromo-8-methyl-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (100 mg, 0.37 mmol), (5-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (147 mg, 0.41 mmol), Cs₂CO₃ (303 mg, 0.93 mmol), and PdCl₂dppf•CH₂Cl₂ (15 mg, 0.019 mmol) were heated in a microwave reactor at 90 °C for 3 hours in 1,4-dioxane (4 mL) and H₂O (2 mL). The reaction mixture was cooled to room temperature, diluted with H₂O (30 mL), and treated with 1 M HCl aqueous solution (1 mL). The solids were collected by filtration, and the filter cake was washed with H₂O. The beige precipitate was dissolved in CH₂Cl₂ / MeOH, dried, and deposited on diatomaceous earth. It was then purified by rapid chromatography (using a CH₂Cl₂ solution with a CH₂Cl₂ / MeOH / concentrated NH₄OH ratio of 89 / 10 / 1) to obtain a white solid, 5-chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (75.0 mg, 40%). MS (ESI) m / z [M+H] + 505.34. 1¹H NMR (500 MHz, DMSO-d⁶) δ ppm 10.10 - 10.81 (m, 1 H), 8.57 - 8.72 (m, 1 H), 8.46 (d, J=2.45 Hz, 1 H), 8.15 (d, J=2.45 Hz, 1 H), 7.84 (d, J=4.40 Hz, 0.7 H), 7.70-7.78 (m, 1.3 H), 7.21 (d, J=4.03 Hz, 1 H), 7.13 - 7.18 (m, 1 H), 7.05 - 7.11 (m, 1 H), 3.96 (s, 3 H), 2.92 (d, J=4.52 Hz, 3 H). One signal corresponding to Me is masked by the solvent peak.

[0283] Example 28: 5-Chloro-N-(2-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-28) Preparation using a general method for SMC: 6-bromo-8-methyl-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (100 mg, 0.37 mmol), (3-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (134 mg, 0.37 mmol), Cs₂CO₃ (303 mg, 0.93 mmol), and PdCl₂dppf•CH₂Cl₂ (15 mg, 0.019 mmol) were heated in a microwave reactor at 90 °C for 3 h in 1,4-dioxane (4 mL) and H₂O (2 mL). The reaction mixture was diluted with H₂O (30 mL) and treated with 1 M HCl aqueous solution (1 mL). The solids were collected by filtration, and the filter cake was washed with H₂O. The beige precipitate was redissolved in CH₂Cl₂ / MeOH, dried, and deposited on diatomaceous earth. It was then purified by rapid chromatography (using a CH₂Cl₂ solution with a CH₂Cl₂ / MeOH / concentrated NH₄OH ratio of 89 / 10 / 1) to obtain a white solid, 5-chloro-N-(2-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (70.0 mg, 37%). MS (ESI) m / z [M+H] + 505.34. 1¹H NMR (500 MHz, DMSO-d⁶) δ ppm 10.25–10.49 (m, 1 H), 8.56–8.76 (m, 1 H), 8.46 (d, J=2.32 Hz, 1 H), 8.06 (d, J=2.57 Hz, 1 H), 7.83 (d, J=4.28 Hz, 0.7 H), 7.68–7.77 (m, 1.3 H), 7.19–7.30 (m, 2 H), 7.13–7.19 (m, 1 H), 3.89 (s, 3 H), 2.92 (d, J=4.40 Hz, 3 H). A signal corresponding to Me is masked by a solvent peak.

[0284] Example 29: N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-29) Step 1: 2-Amino-6-bromo-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one To a suspension of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (511 mg, 1.61 mmol) in i-PrOH (15 mL), NH3 (7 N MeOH solution, 4.6 mL, 32 mmol) was added at room temperature. The mixture was shaken at room temperature for 1.2 h. The reaction mixture was diluted with 1 M KOH aqueous solution (10 mL, 10 mmol), shaken for a few minutes, and left to stand overnight at room temperature. The yellow precipitate was collected by filtration and washed with H2O to give a yellow solid of 2-amino-6-bromo-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one (189.0 mg, 44%). MS (ESI) m / z [M+H] + 255.10|257.13.

[0285] Step 2: N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-29) SMC was prepared using a general method: 5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (85 mg, 0.23 mmol), 2-amino-6-bromo-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (60 mg, 0.23 mmol), Cs₂CO₃ (192 mg, 0.59 mmol), and PdCl₂dppf (17 mg, 0.024 mmol) were heated in a microwave reactor at 90 °C for 2 hours in H₂O (2 mL) and 1,4-dioxane (4 mL). The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1, to give a white solid N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (48 mg, 42% yield). MS (ESI) m / z [M+H + 491.20. 1 HNMR (500 MHz, DMSO-d6) δ ppm 10.38 (br. s, 1 H), 8.62 (s, 1 H), 8.45 (d, J=2.57 Hz, 1 H), 8.14 (d, J=2.57 Hz, 1 H), 7.74 (s, 1 H), 7.35 (s, 2 H), 7.19 (dd, J=6.36, 2.57 Hz, 1 H), 7.10 - 7.17 (m, 1 H), 7.04 - 7.10 (m, 1 H), 3.95 (s, 3 H). The signal corresponding to “N-Me” appears to be masked by the “DMSO” peak.

[0286] Example 30: N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-30) Preparation using the universal SMC method: 2-amino-6-bromo-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (60 mg, 0.23 mmol), (3-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (85 mg, 0.23 mmol), Cs₂CO₃ (192 mg, 0.59 mmol), and PdCl₂dppf (17 mg, 0.024 mmol) were heated in a microwave reactor at 90 °C for 2 hours in H₂O (2 mL) and 1,4-dioxane (4 mL). The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (12 g SiO₂RediSep GOLD® column, using a CH₂Cl₂ / MeOH / concentrated NH₄OH solution at 89 / 10 / 1). The collected solid was then dissolved in MeOH, filtered through a Waters PoraPak CX column, washed with MeOH, and eluted with a 1.4 M NH3 MeOH solution to give a white solid N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (33.4 mg, 29% yield). MS (ESI) m / z [M+H] + 491.20. 1 H NMR (500 MHz, DMSO-d6) δ ppm10.35 (br. s., 1 H), 8.62 (s, 1 H), 8.48 (d, J=1.83 Hz, 1 H), 8.06 (d, J=2.57Hz, 1 H), 7.66 - 7.80 (m, 1 H), 7.38 (s, 2 H), 7.05 - 7.30 (m, 3 H), 3.82 -3.98 (m, 3 H), 3.49 - 3.58 (m, 3 H).

[0287] Example 31: 5-Chloro-N-(5-fluoro-4-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide (I-31) Step 1: 5-Chloro-N-(5-fluoro-4-iodopyridin-2-yl)-2-methoxypyridin-3-sulfonamide 5-Fluoro-4-iodopyridin-2-amine (0.70 g, 2.9 mmol) was added to anhydrous pyridine (7 mL) at 0 °C, and the reaction mixture was stirred at room temperature for 2 h. Then, 5-chloro-2-methoxypyridin-3-sulfonyl chloride (0.70 g, 2.9 mmol) was added at 0 °C, and the reaction mixture was stirred at room temperature for 2 h. Another batch of 5-chloro-2-methoxypyridin-3-sulfonyl chloride (1 equivalent) was added at 0 °C, and stirring was continued at room temperature for 16 h. The reaction mixture was then diluted with 10% citric acid solution (40 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were dried (anhydrous Na₂SO₄) and concentrated under reduced pressure. The white solid 5-chloro-N-(5-fluoro-4-iodopyridin-2-yl)-2-methoxypyridin-3-sulfonamide (0.70 g, 55 %) was obtained by column chromatography purification using a hexane solution of EtOAc. 1 H NMR (400 MHz, DMSO-d6) δ 11.47 (s, 1H), 8.49 (d, J=2.0 Hz, 1H), 8.25 (d, J=2.0 Hz, 1H), 8.07 (s, 1H), 7.49 (d, J = 3.6Hz, 1H), 3.88 (s, 3H). MS (ESI)m / z [M+H] + 443.9.

[0288] Step 2: (8-Methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid Preparation using the universal method MB: 6-bromo-8-methyl-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (140 mg, 0.52 mmol), B2pin2 (396 mg, 1.56 mmol), KOAc (179 mg, 1.82 mmol), and PdCl2dppf (38 mg, 0.052 mmol) were heated in an oil bath at 100 °C for 4 hours in anhydrous 1,4-dioxane (10 mL). The crude reaction mixture (0.052 M) was used directly for the next step. MS (ESI) m / z [M+H] + 235.23.

[0289] Step 3: 5-Chloro-N-(5-fluoro-4-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide (I-31) SMC was prepared using a general method: 5-chloro-N-(5-fluoro-4-iodopyridin-2-yl)-2-methoxypyridin-3-sulfonamide (80 mg, 0.18 mmol), PdCl2dppf (13.2 mg, 0.018 mmol), Cs2CO3 (176 mg, 0.54 mmol), and crude product (8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid (4.2 mL, 0.22 mmol, 0.052 M 1,4-dioxane solution) were heated in a microwave reactor at 65 °C for 3 hours, followed by heating at 90 °C for 3 hours. The reaction mixture was concentrated under reduced pressure and deposited on diatomaceous earth. It was purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 to obtain a solid. This solid was ground with Et2O to give a white solid, 5-chloro-N-(5-fluoro-4-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide (13.4 mg, 15% yield). MS (ESI) m / z [M+H] + 506.17. 1 H NMR (500 MHz, DMSO-d6) δ ppm 11.37 (br. s., 1 H), 8.59 - 8.80 (m, 1 H), 8.50 (d, J=2.32 Hz, 1 H), 8.28 (d, J=2.57 Hz, 1 H), 8.19 (s, 1 H), 8.02 (d, J=4.77 Hz, 0.65 H), 7.99 (s, 1 H), 7.91 (d, J=4.52 Hz, 0.35 H), 7.24 (br. s., 1 H), 3.90 (s, 3 H), 3.61 (s, 2 H), 3.53 (s, 1 H), 2.86 - 2.96 (m, 3H).

[0290] Example 32: 5-Chloro-N-(3-fluoro-4-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide (I-32) Step 1: 5-Chloro-N-(3-Fluoro-4-iodopyridin-2-yl)-2-methoxypyridin-3-sulfonamide To a cooled solution of t-BuOK (0.7 g, 6.3 mmol) in THF (5 mL), 3-fluoro-4-iodopyridin-2-amine (0.50 g, 2.1 mmol) was added at 0 °C. Stirring was continued at room temperature for 1 hour, followed by the addition of 5-chloro-2-methoxypyridin-3-sulfonyl chloride (0.51 g, 2.1 mmol) at 0 °C. After stirring at room temperature for 4 hours, the reaction mixture was diluted with H₂O (5 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were dried (anhydrous Na₂SO₄) and concentrated under reduced pressure. Purification by column chromatography with hexane and EtOAc yielded a white solid, 5-chloro-N-(3-fluoro-4-iodopyridin-2-yl)-2-methoxypyridin-3-sulfonamide (0.3 g, 33 %). 1 H NMR (400 MHz, DMSO-d6) δ 11.58 (br.s., 1H), 8.48 (s, 1H), 8.22 (s, 1H), 7.60 (s, 1H), 7.48 (br.s., 1H), 3.86 (s, 3H). MS(ESI)m / z [M+H] + 444.1.

[0291] Step 2: (2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid Preparation using the universal method MB: 6-bromo-2-(methylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (150 mg, 0.59 mmol), B2pin2 (224 mg, 0.88 mmol), KOAc (202 mg, 2.06 mmol), and PdCl2dppf (55.9 mg, 0.076 mmol) were heated in an oil bath at 105 °C for 1 day and 19 hours in anhydrous 1,4-dioxane (24 mL). The crude mixture was estimated to contain 84% (2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid. The reaction mixture was used as a crude product for the next step. MS (ESI) m / z [M+H] + 221.24.

[0292] Step 3: 5-Chloro-N-(3-fluoro-4-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide (I-32) SMC was prepared using a general method: 5-chloro-N-(3-fluoro-4-iodopyridin-2-yl)-2-methoxypyridin-3-sulfonamide (80 mg, 0.18 mmol), PdCl2dppf (13 mg, 0.018 mmol), Cs2CO3 (176 mg, 0.54 mmol) in H2O (4.3 mL) and crude product (2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)boronic acid in 1,4-dioxane (8.7 mL, 0.22 mmol) was heated in a microwave reactor at 60 °C for 2.5 h in a sealed environment, followed by heating at 100 °C for 2 h. The solution was purified by rapid chromatography using a CH2Cl2 solution (0-100%) with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1, followed by purification with a MeOH CH2Cl2 solution to obtain a brown solid. This solid was ground with Et2O and then repeatedly ground with MeCN until a gray solid of 5-chloro-N-(3-fluoro-4-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide (30.0 mg, 33% yield based on 97% purity) was obtained. MS (ESI) m / z [M+H] + 492.22. 1 H NMR (500 MHz, DMSO-d6) δ ppm 11.97 - 12.32 (m,1 H), 8.54 - 8.76 (m, 1 H), 8.45 (br. s., 1 H), 8.22 (br. s., 1 H), 7.96 (br.s., 1 H), 7.87 (br.s., 1.5 H), 7.56 - 7.72 (m, 0.5 H), 6.84 - 7.30 (m, 2 H), 3.74 - 4.03 (br.s., 3 H), 2.81 - 2.96 (m, 3 H).

[0293] Example 33: (trans-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (trans I-33) Step 1: trans-4-aminocyclohexane-1-carboxamide trifluoroacetic acid A mixture of TBTU (693 mg, 2.2 mmol), trans-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (500 mg, 2.0 mmol), and DIPEA (1.1 mL, 6.2 mmol) in DMF (3 mL) was stirred at room temperature for 30 minutes, followed by the addition of solid NH4Cl (220 mg, 4.1 mmol). The reaction mixture was stirred overnight at room temperature, then diluted with Et2O and washed with 0.1 M HCl aqueous solution. The organic layer was separated and concentrated under reduced pressure to give a white solid (100 mg). Another batch of white solid was collected by filtering the aqueous layer. The filter cake was washed with excess H2O. The two batches were combined to give tert-butyl (trans-4-carbamoylcyclohexyl)carbamate (476.0 mg, 96%). All materials were stirred in CH2Cl2 (5 mL) and TFA (0.16 mL, 2.0 mmol) at room temperature for 1 day and 18 hours. The reaction mixture was then concentrated under reduced pressure and dried under vacuum to obtain a light yellow gel, trans-4-aminocyclohexane-1-carboxamide trifluoroacetic acid (505 mg, 96%), which was used directly in the next step.

[0294] Step 2: trans-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide A mixture of trans-4-aminocyclohexane-1-carboxamide trifluoroacetic acid (168 mg, 0.66 mmol) and K₂CO₃ (453 mg, 3.3 mmol) in DMF (4 mL) was stirred at room temperature for 0.2 h. 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (217 mg, 0.68 mmol) was added in a single addition, and stirring was continued at room temperature for 2.5 h, followed by the addition of H₂O (4 mL). The solvent was removed by vacuum evaporation, and the residue was redissolved in MeOH, deposited on diatomaceous earth, and purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1, followed by purification with a MeOH-CH2Cl2 solution to obtain a white solid trans-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (216 mg, purity 77%). MS (ESI) m / z [M+H + 380.20|382.16.

[0295] Step 3: trans-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (trans I-33) SMC was prepared using a general method: 5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (80 mg, 0.22 mmol), Cs₂CO₃ (180 mg, 0.55 mmol), trans-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (109 mg, 0.22 mmol, 77 %) and PdCl₂dppf (16 mg, 0.022 mmol) were heated in a microwave reactor at 90 °C for 2 hours in 1,4-dioxane (4 ml) and H₂O (2 ml). Purification by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 gave a white solid, trans-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (38.0 mg, 28% yield based on 99.7% purity). MS (ESI) m / z [M+H + 616.21. 1 H NMR (500 MHz, DMSO-d6) δ = 10.58 - 10.36 (m, 1H), 8.70 - 8.59 (m, 1H), 8.47 (d, J = 2.4 Hz, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.91 (br d, J = 7.7 Hz, 1H), 7.79 - 7.71 (m, 1H), 7.25 - 7.18 (m, 2H), 7.18 - 7.12 (m, 1H), 7.11 - 7.05 (m, 1H), 6.72 - 6.64(m, 1H),, 3.96 (s, 3H), 3.90 -3.73 (m, 1H), 3.60 - 3.51 (m, 3H), 2.14 - 1.74 (m, 5H), 1.54 - 1.21 (m, 4H).

[0296] Example 34: cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (cis I-33) Step 1: cis-4-aminocyclohexane-1-formamide (TFA) A mixture of TBTU (693 mg, 2.2 mmol) and cis-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (500 mg, 2.0 mmol) in DMF (3 mL) was treated with DIPEA (1.1 mL, 6.2 mmol). After stirring at room temperature for 30 minutes, solid NH4Cl (220 mg, 4.1 mmol) was added. The reaction mixture was stirred overnight at room temperature, then diluted with Et2O and washed with 0.1 M HCl aqueous solution. The organic layer was concentrated under reduced pressure to give an off-white solid, which was dissolved in CH2Cl2 (5 mL) and TFA (0.79 mL, 10 mmol) and stirred at room temperature for 1 day and 18 hours. The reaction mixture was concentrated under reduced pressure and dried under vacuum to give an off-white solid of cis-4-aminocyclohexane-1-carboxamide trifluoroacetate (318 mg, 60%).

[0297] Step 2: cis-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide A mixture of cis-4-aminocyclohexane-1-carboxamide*TFA (175 mg, 0.68 mmol) and K₂CO₃ (472 mg, 3.4 mmol) in DMF (3 mL) was shaken at room temperature for 0.2 h. Then, 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (120 mg, 0.38 mmol) was added in a single batch. Stirring was continued at room temperature for 2.5 h. The mixture was then diluted with water and filtered to give a white solid cis-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (109 mg, 41% yield based on 54% purity). MS (ESI) m / z [M+H] + 380.20|382.16.

[0298] Step 3: cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (cis I-33) SMC was prepared using a general method: 5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (114 mg, 0.31 mmol), Cs₂CO₃ (233 mg, 0.72 mmol), cis-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (109 mg, 0.15 mmol, 54 %), and PdCl₂dppf (21 mg, 0.029 mmol) were heated at 100 °C for 2.5 h in 1,4-dioxane (6 mL) and H₂O (3 mL). Purification by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 yielded a beige solid, cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (42.0 mg, 42% yield based on 97% purity). MS (ESI) m / z [M+H + 616.29. 1 H NMR (500 MHz, DMSO-d6) δ = 10.64 - 10.34(m, 1H), 8.69 - 8.62 (m, 1H), 8.48 (d, J = 2.4 Hz, 1H), 8.19 - 8.14 (m, 1H),7.91 - 7.84 (m, 1H),7.75 (br s, 1H), 7.24 - 7.12 (m, 3H), 7.11 - 7.04 (m,1H), 6.76 - 6.64 (m, 1H), 4.08 - 3.98 (m, 1H), 3.96 (s, 3H), 3.56 (s, 3H),,2.26 - 2.17 (m, 1H), 1.98 - 1.46 (m, 8H).

[0299] Example 35: trans-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (trans I-34) Step 1: trans-4-amino-N-methylcyclohexane-1-carboxamide, trifluoroacetic acid A DMF (3 mL) suspension of TBTU (693 mg, 2.2 mmol) and trans-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (500 mg, 2.0 mmol) was treated with DIPEA (1.074 mL, 6.17 mmol). After stirring at room temperature for 10 min, solid MeNH2*HCl (166 mg, 2.5 mmol) was added. The reaction mixture was shaken at room temperature for 30 min, then stirred at room temperature for 2 days. The reaction mixture was then diluted with Et2O and washed with 0.1 M HCl aqueous solution. The organic layer was concentrated under reduced pressure. Another batch of off-white solid was collected by filtering the aqueous layer. The filtered solid was washed with excess H2O, combined with the material from the Et2O layer, and dried to give a white solid trans-4-(methylcarbamoyl)cyclohexyl)carbamate tert-butyl ester (417 mg, 79% yield). MS (ESI) m / z [M+H] + 257.36. All materials were stirred in CH2Cl2 (5 mL) and TFA (0.71 mL, 9.2 mmol) at room temperature for 2 days. The reaction was concentrated under reduced pressure. The material was sonicated with Et2O (5 mL), left to stand overnight at room temperature, and then filtered. The filter cake was washed with excess Et2O to give a white solid trans-4-amino-N-methylcyclohexane-1-carboxamide trifluoroacetate (370.0 mg, 67%). MS (ESI) m / z [M+H] + 157.33.

[0300] Step 2: trans-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide A mixture of trans-4-amino-N-methylcyclohexane-1-carboxamide trifluoroacetate (0.102 g, 0.38 mmol) and K₂CO₃ (0.217 g, 1.6 mmol) in DMF (5 mL) was shaken at room temperature for 30 minutes, followed by the addition of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (0.100 g, 0.31 mmol) in a single batch. The reaction mixture was then shaken for 2 hours and stirred at room temperature for 5 days. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 to give a white solid trans-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (39.0 mg, 32% yield). MS (ESI) m / z [M+H + 394.20|396.23.

[0301] Step 3: trans-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (trans I-34) A mixture of (5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (46.4 mg, 0.13 mmol), Cs₂CO₃ (81 mg, 0.25 mmol), trans-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (39 mg, 0.099 mmol) and PdCl₂dppf (7 mg, 0.010 mmol) in 1,4-dioxane (6 mL) and H₂O (3 mL) was degassed with an argon stream. The reaction was then heated at 100 °C for 2.1 h. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1, to give a white solid trans-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (29 mg, 46% yield based on 98% purity). MS (ESI) m / z [M+H + 630.28.1 H NMR (500 MHz, DMSO-d6) δ = 10.62 - 10.37 (m, 1H), 8.70 -8.61 (m, 1H), 8.48 (d, J = 2.4 Hz, 1H), 8.16 (d, J = 2.6 Hz, 1H), 7.91 (br d, J = 7.6 Hz, 1H), 7.79 - 7.72 (m, 1H), 7.68 (br s, 1H),, 7.25 - 7.19 (m, 1H), 7.18 - 7.12 (m, 1H), 7.11 - 7.04 (m, 1H), 3.96 (s, 3H), 3.90 - 3.72 (m, 1H),3.63 - 3.50 (m, 3H), 2.57 (br d, J = 4.3 Hz, 3H), 2.14 - 1.88 (m, 3H), 1.86 -1.72 (m, 2H), 1.58 - 1.40 (m, 2H), 1.38 - 1.22 (m, 2H).

[0302] Example 36: (cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (cis I-34) Step 1: cis-4-amino-N-methylcyclohexane-1-carboxamide, trifluoroacetic acid A DMF (3 mL) suspension of TBTU (693 mg, 2.2 mmol) and cis-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (500 mg, 2.0 mmol) was treated with DIPEA (1.1 mL, 6.2 mmol). After stirring at room temperature for 10 min, solid MeNH2*HCl (166 mg, 2.5 mmol) was added. The reaction mixture was shaken at room temperature for 30 min, then stirred overnight at room temperature, diluted with Et2O, and then washed with 0.1 M HCl aqueous solution. The organic layer was concentrated under reduced pressure to give a pale yellow oil (cis-4-(methylcarbamoyl)cyclohexyl)carbamate (640.0 mg). The entire mixture was stirred overnight at room temperature in CH2Cl2 (5 mL) and TFA (0.71 mL, 9.2 mmol). The reaction solution was concentrated under reduced pressure and then briefly dried under high vacuum to obtain a white solid, which was suspended in Et2O (5 mL) and sonicated. The mixture was aged overnight at room temperature, and the white precipitate was collected by filtration. The filter cake was washed with excess Et2O to obtain a colorless colloidal substance, cis-4-amino-N-methylcyclohexane-1-carboxamide trifluoroacetate (472.0 mg, 85%).

[0303] Step 2: cis-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide Cis-4-amino-N-methylcyclohexane-1-carboxamide trifluoroacetate (0.102 g, 0.38 mmol) and K₂CO₃ (0.22 g, 1.6 mmol) were shaken in DMF (5 mL) at room temperature for 30 minutes. 6-Bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (0.100 g, 0.31 mmol) was added in a single dose, and shaking was continued at room temperature for 2 hours. The reaction mixture was then stirred overnight at room temperature, concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 to give a white solid (1S,4S)-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (49.0 mg, 38% yield based on 96% purity). MS (ESI) m / z [M+H + 394.20|396.23.

[0304] Step 3: (cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (cis I-34) SMC was prepared using a general method: (5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (56 mg, 0.15 mmol), Cs₂CO₃ (97 mg, 0.30 mmol), (cis-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (49 mg, 0.12 mmol), and PdCl₂dppf (8.7 mg, 0.012 mmol) were heated at 100 °C for 2 hours under argon in 1,4-dioxane (6 mL) and H₂O (3 mL). The analytes were analyzed by rapid chromatography using CH₂Cl₂ / MeOH / concentrated NH₄OH. Purification with CH2Cl2 solution at 89 / 10 / 1 yielded a white solid (cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (41 mg, 53% yield based on 98% purity). MS (ESI) m / z [M+H + 630.21. 1 H NMR (500 MHz, DMSO-d6) δ =10.57 - 10.39 (m, 1H), 8.70 - 8.61 (m, 1H), 8.48 (d, J = 2.4 Hz, 1H), 8.16(d, J = 2.6 Hz, 1H), 7.88 (br d, J = 6.2 Hz, 1H), 7.79 - 7.70 (m, 1H), 7.67 -7.58 (m, 1H), 7.24 - 7.19 (m, 1H), 7.18 - 7.12 (m, 1H), 7.11 - 7.05 (m, 1H), 4.10 - 4.00 (m, 1H), 3.97 (s, 3H), 3.56 (s, 3H), 2.58 (d, J= 4.5 Hz, 3H), 2.28 - 2.19 (m, 1H), 1.95 - 1.83 (m, 3H), 1.82 - 1.72 (m, 1H), 1.70 - 1.58 (m, 2H), 1.57 - 1.47 (m, 2H).

[0305] Example 37. Trans-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclobutane-1-carboxamide (trans I-35) Step 1: trans-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclobutane-1-carboxamide Trans-aminocyclobutane-1-carboxylate (36 mg, 0.24 mmol) and 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (50 mg, 0.16 mmol) were treated in NMP (3 mL) with DIPEA (0.14 mL, 0.79 mmol) and stirred at room temperature for 2 days and 20 hours. MS (ESI) m / z [M+H] + 353.1. MeNH2 (33 wt% EtOH solution, 0.20 mL, 1.6 mmol) was added to the crude reaction mixture, and the reaction mixture was cooled to 0 °C. Solid TBTU (96 mg, 0.30 mmol) was added, and the reaction was stirred while cooling, then stirred at room temperature for a total of 1 day and 4 hours. MS (ESI) m / z [M+H] + 366.21|368.17. The reaction mixture was diluted with water (6 mL) and then concentrated under reduced pressure to remove most of the volatiles.

[0306] Step 2: trans-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclobutane-1-carboxamide (trans I-35) Preparation using the general method SMC: using the crude product (1r,3r)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclobutane-1-carboxamide from the previous step in NMP (2 mL), PdCl2dppf (11 mg, 0.016 mmol), Cs2CO3 (154 mg, 0.47 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (79 mg, 0.22 mmol) in 1,4-dioxane (6 mL) and H2O (3 mL), and heated in a sealed container at 90 °C for 20 hours. The product was purified twice by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1, yielding a pale yellow solid, trans-3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclobutane-1-carboxamide (30.0 mg, yield 31% based on 97% purity). MS (ESI) m / z [M+H + 602.29. 1 H NMR (500 MHz, DMSO-d6) δ ppm 10.37 (br. s., 1 H), 8.60 - 8.68 (m, 1 H), 8.49 (d, J=2.69 Hz, 1 H), 8.30 (d, J=6.85 Hz, 0.7 H), 8.16 (d, J=7.46 Hz, 0.3H), 8.07 (d, J=2.57 Hz, 1 H), 7.70 - 7.77 (m, 2 H), 7.13 - 7.31 (m, 3 H), 3.90 (s, 3 H), 3.50 - 3.58 (m, 3 H), 3.29 - 3.34 (m, 1 H), 2.83 - 2.96 (m, 1H), 2.60 (d, J=4.65 Hz, 3 H), 2.16 - 2.30 (m, 3 H), 1.83 - 1.98 (m, 1 H).

[0307] Example 38. Racemic-(1R,3S)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide((racemic-(1R,3S) I-36) Step 1: Racemic-(1R,3S)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid A mixture of cis-3-aminocyclohexanecarboxylic acid (500 mg, 3.5 mmol), di-tert-butyl dicarbonate (915 mg, 4.2 mmol), and NaHCO3 (880 mg, 10 mmol) in THF (6 mL) and H2O (6 mL) was stirred at 0 °C and slowly heated to room temperature. The reaction was then held as a suspension at room temperature and stirred for 1 day and 18 hours. The reaction was then extracted with Et2O. The aqueous phase was carefully acidified to pH 4–5 with 1 M citric acid solution, producing a white precipitate, which was collected by filtration. The solid was washed with a small amount of Et2O and then dried to give a white solid racemic-(1R,3S)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (783.0 mg, 93%). MS (ESI) m / z [M+H] + 244.27.

[0308] Step 2: Racemic-(1R,3S)-3-amino-N-methylcyclohexane-1-carboxamide TFA salt Racemic-(1R,3S)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (200 mg, 0.82 mmol) and TBTU (317 mg, 0.99 mmol) were sonicated in CH2Cl2 (10 mL). DIPEA (0.29 mL, 1.6 mmol) was added, and the reaction mixture was shaken at room temperature for several minutes. Then, MeNH2 (33 wt.% EtOH solution, 0.31 mL, 2.5 mmol) was added in a single addition at room temperature. Shaking was continued at room temperature for 1.5 hours. The evaporation product was evaporated under reduced pressure to give crude racemic-((1R,3S)-3-(methylcarbamoyl)cyclohexyl)carbamate tert-butyl. MS (ESI) m / z [M+H] + 257.29.

[0309] The residue was then shaken in CH2Cl2 (6 mL) and TFA (1.0 mL, 13 mmol) at room temperature for 1.5 h. The reaction mixture was then left to stand overnight at -20 °C. The mixture was concentrated under reduced pressure and dried under vacuum to give crude racemic-(1R,3S)-3-amino-N-methylcyclohexane-1-carboxamide TFA salt as a colorless gel, which was used directly in the next step without further purification.

[0310] Step 3: Racemic-(1R,3S)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide((racemic-(1R,3S) I-36) K₂CO₃ (373 mg, 2.7 mmol) and crude racemic-(1R,3S)-3-amino-N-methylcyclohexane-1-carboxamide TFA salt (73 mg, 0.27 mmol) were shaken in NMP (1.5 mL) at room temperature for 10 minutes. Then, solid 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (95 mg, 0.297 mmol) was added at room temperature. The mixture was stirred overnight at room temperature for a total of 3 days and 20 hours. After the reaction was complete, the entire crude product containing racemic-(1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide was mixed with MeOH and concentrated under reduced pressure. MS (ESI)m / z [M+H] + 394.27|396.23.

[0311] Then, using all the crude racemic-(1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide remaining in NMP, it was purified by general method SMC using Pd(dppf)Cl2 (19.79 mg, 0.027 mmol), Cs2CO3 (264 mg, 0.810 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (136 mg, 0.38 mmol), H2O (3 mL), and 1,4-dioxane (6 mL) at 100 °C for 2.4 h. The purified product was purified by rapid chromatography (SiO2 was purified using a CH2Cl2 solution with CH2Cl2 / MeOH / concentrated NH4OH at a concentration of 89 / 10 / 1). The collected solid material was ultrasonically ground with Et2O to obtain a white solid racemic-(1R,3S)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (10.4 mg, yield 6% based on 95% purity). Yields are reported as the total yield from all steps starting with racemic-(1R,3S)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid. MS (ESI) m / z [M+H] + 630.43.

[0312] 1 H NMR (500 MHz, DMSO) δ 10.36 (s, 1H), 8.70 -- 8.59 (m, 1H), 8.47(s, 1H), 8.07 (d, J = 2.4 Hz, 1H), 7.95 (d, J = 7.8 Hz, 0.6H), 7.82 (d, J =7.3 Hz, 0.4 H), 7.76 -- 7.68 (m, 2H), 7.32 -- 7.09 (m, 3H), 3.98 -- 3.82 (m,4H), 3.59 -- 3.50 (m, 3H), 2.58 -- 2.54 (m, 3H), 2.31 -- 2.21 (m, 1H), 2.02-- 1.75 (m, 2H), 1.75-1.65 (m, 1H), 1.48 -- 1.18 (m, 5H).

[0313] Example 39. Racemic-(1R,3R)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide((racemic-(1R,3R) I-36) Step 1: Racemic-(1R,3R)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid A mixture of di-tert-butyl dicarbonate (729 mg, 3.3 mmol), racemic-(1R,3R)-3-aminocyclohexane-1-carboxylic acid salt (500 mg, 2.8 mmol), and NaHCO3 (701 mg, 8.35 mmol) in THF (6 mL) and H2O (6 mL) was stirred at 0 °C and slowly heated to room temperature. The reaction mixture was then stirred as a suspension at room temperature for 1 day and 18 hours. The reaction was then extracted with Et2O. The aqueous phase was carefully acidified to pH 4–5 with 1 M citric acid solution and extracted with DCM (3x). The combined organic extracts were concentrated under reduced pressure and dried to give racemic-(1R,3R)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (138 mg, 20%) as a white solid. MS (ESI) m / z [M+H] + 244.27.

[0314] Step 2: Racemic-(1R,3R)-3-amino-N-methylcyclohexane-1-carboxamide, TFA Racemic-(1R,3R)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (138 mg, 0.57 mmol) and TBTU (219 mg, 0.68 mmol) were sonicated in CH2Cl2 (6 mL). DIPEA (0.198 mL, 1.134 mmol) was added, and the reaction mixture was shaken at room temperature for several minutes. Then, MeNH2 (33 wt.% EtOH solution, 0.21 mL, 1.7 mmol) was added in a single addition at room temperature. Shaking was continued at room temperature for 1.5 hours. The evaporation product was evaporated under reduced pressure to give crude racemic-((1R,3R)-3-(methylcarbamoyl)cyclohexyl)carbamate tert-butyl. MS (ESI) m / z [M+H] + 257.36.

[0315] The residue was then shaken in CH2Cl2 (6 mL) and TFA (0.7 mL, 9 mmol) at room temperature for 1.5 h. The reaction mixture was then left to stand overnight at -20 °C. The mixture was concentrated under reduced pressure and dried under vacuum to give crude racemic-(1R,3S)-3-amino-N-methylcyclohexane-1-carboxamide, TFA, as a colorless gel, which was used directly in the next step without further purification.

[0316] Step 3: Racemic-(1R,3R)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide((racemic-(1R,3R) I-36) The crude racemic-(1R,3S)-3-amino-N-methylcyclohexane-1-carboxamide TFA salt (74 mg, 0.27 mmol) and K₂CO₃ (378 mg, 2.7 mmol) were shaken in NMP (1.5 mL) at room temperature for 15 min, followed by the addition of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (96 mg, 0.30 mmol) at room temperature. The mixture was stirred at room temperature for 3 days and 20 hours. The entire crude product containing racemic-(1R,3R)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide was diluted with MeOH and concentrated under reduced pressure. MS (ESI) m / z [M+H] +394.27|396.23. Then, using all the crude racemic-(1R,3R)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide from the residual NMP, according to the general method SMC, using Pd(dppf)Cl2 (20 mg, 0.027 mmol), Cs2CO3 (268 mg, 0.82 mmol) and (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (138 mg, 0.38 mmol), H2O (3 mL) and 1,4-dioxane (6 mL), heated at 100 °C for 2.3 h. Purification by rapid chromatography (SiO2 using CH2Cl2 solution with CH2Cl2 / MeOH / concentrated NH4OH at a ratio of 89 / 10 / 1) yielded a white solid racemic-(1R,3R)-3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (11.2 mg, yield 6% based on 99% purity). MS (ESI) m / z [M+H + 630.43. 1 H NMR (500 MHz, DMSO-d 6) δ ppm 10.36(br. s., 1 H), 8.58 - 8.72 (m, 1 H), 8.48 (d, J =2.69 Hz, 1 H), 8.07 (d, J =2.57 Hz, 1 H), 7.79 (d, J =7.21 Hz, 0.6 H), 7.75 (s, 1 H), 7.67-7.59 (m, 0.4H), 7.53 - 7.60 (m, 1 H), 7.12 - 7.31 (m, 3 H), 4.14 - 4.33 (m, 1 H), 3.90 (s, 3 H), 3.55 (s, 3 H), 2.56 (d, J =4.52 Hz, 3 H), 1.47 - 1.89 (m, 9 H).

[0317] Example 40. Cis-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide (cis-I-37) Step 1: (1S,3S)-3-aminocyclobutane-1-carboxamide, trifluoroacetic acid A suspension of cis-3-aminocyclobutane-1-carboxylic acid (200 mg, 1.74 mmol) and Boc₂O (398 mg, 1.82 mmol) in CH₂Cl₂ (10 mL) and DIPEA (0.91 mL, 5.21 mmol) was sonicated and then stirred at room temperature for 3 days and 21 hours. TBTU (586 mg, 1.824 mmol) was added at room temperature, and the reaction was shaken at room temperature for 5 minutes, followed by the addition of 4-methoxybenzylamine (0.24 mL, 1.8 mmol). Stirring was continued at room temperature for 21 hours. The white solid was collected by filtration, washed with DCM, and dried under vacuum to give a white solid (cis-3-((4-methoxybenzyl)carbamoyl)cyclobutyl)carbamate (432.0 mg, 54% yield based on 72% purity). MS (ESI) m / z [M+H] + 335.37. Tert-butyl (cis-3-((4-methoxybenzyl)carbamoyl)cyclobutyl)carbamate (209 mg, 0.62 mmol) was dissolved in TFA (1.0 mL, 13.7 mmol) and stirred at room temperature for 5 days and 20 hours. The reaction mixture was then heated overnight at 60 °C. The solvent was removed under reduced pressure, followed by high vacuum treatment to obtain a red gel, all of which was used in the next step.

[0318] Step 2: cis-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide A DMF (3 mL) suspension of cis-3-aminocyclobutane-1-carboxamide trifluoroacetate (143 mg, 0.63 mmol) (from all residues in the previous step) and K₂CO₃ (346 mg, 2.507 mmol) was shaken at room temperature for 10 minutes, followed by the addition of 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (219 mg, 0.69 mmol) in a single batch. Stirring was continued at room temperature for 3 days and 17 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 to give a beige solid cis-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide (68.0 mg, 30% yield based on 97% purity). MS (ESI) m / z [M+H + 352.22|354.17.

[0319] Step 3: cis-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide (cis-I-37) SMC was prepared according to the general method: cis-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide (68 mg, 0.18 mmol), Cs2CO3 (153 mg, 0.47 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (88 mg, 0.24 mmol) and Pd(dppf)Cl2*CH2Cl2 (15 mg, 0.019 mmol) in 1,4-dioxane (4 mL) and H2O (2 mL) was heated at 100 °C for 2.5 h in a microwave reactor. Purification by rapid chromatography using a CH2Cl2 solution with a CH2Cl2 / MeOH / concentrated NH4OH ratio of 89 / 10 / 1 gave a beige solid (cis-3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide (42.0 mg, 38% yield). MS (ESI) m / z [M+H] + 588.38. 1H NMR (500 MHz, DMSO) δ 8.68 -- 8.59(m, 1H), 8.48 (s, 1H), 8.23 ​​(d, J = 6.3 Hz, 0.7H), 8.11 (d, J = 5.9 Hz,0.3H), 8.06 (s, 1H), 7.74 (s, 1H), 7.31 -- 7.20 (m, 3H), 7.17 (t, J = 7.7 Hz,1H), 6.76 (s, 1H), 4.45 -- 4.34 (m, 0.5H), 4.33 -- 4.24 (m, 1H), 4.16 (q, J =5.0 Hz, 0.5H), 3.89 (s, 3H), 3.62 -- 3.50 (m, 3H), 2.78 -- 2.62 (m, 1H), 2.47 -- 2.31 (m, 1H), 2.22 -- 2.06 (m, 2H).

[0320] Example 41: ((5-chloro-2-methoxypyridin-3-yl)sulfonyl)(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)aminopotassium (I-38 K salt) 5-Chloro-N-(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (272 mg, 0.55 mmol) was suspended in EtOH (80 mL) and sonicated. KOH aqueous solution (0.5 M, 1.11 mL, 0.55 mmol) was added dropwise at room temperature. The reaction was sonicated and diluted with H2O (70 mL). After repeated brief sonication, excess EtOH was removed under reduced pressure, and the remaining material was freeze-dried to obtain a light yellow, light powder of ((5-chloro-2-methoxypyridin-3-yl)sulfonyl)(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)amino potassium (291.0 mg, 99%). MS (ESI)m / z [M+H] + 491.4. 1HNMR(500 MHz, DMSO-d6) δ 8.62 -- 8.44 (m, 1H), 8.11 (s, 1H), 7.91 (s, 1H), 7.59 (s, 1H), 7.53 (brs, 0.7H), 7.35 (brs, 0.3H), 6.98 (t, J = 8.1 Hz, 1H), 6.69 (t, J = 7.6 Hz, 1H), 6.46 (t, J = 6.1 Hz, 1H), 3.75 (s, 3H), 2.80 (d, J = 3.6 Hz, 3H). 19 F NMR (471 MHz, DMSO-d6) δ -128.40, -128.53.

[0321] Example 42: ((5-chloro-2-methoxypyridin-3-yl)sulfonyl)(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)aminopotassium (I-25 K salt) 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (374.6 mg, 0.742 mmol) was suspended in EtOH (110 mL) and H₂O (100 mL) and sonicated. Aqueous KOH solution (0.5 M, 1.52 g, 0.74 mmol) was added dropwise at room temperature. After sonication, excess EtOH was removed under reduced pressure, and the water sample was freeze-dried to obtain a light yellow powder (361 mg, 89%). MS (ESI) m / z [M+H] + 505.16.

[0322] 1 H NMR(500 MHz, DMSO-d6) δ 11.76 (brs, 1H), 8.66 -- 8.48 (m, 1H), 8.17(s, 1H), 7.97 (s, 1H), 7.64 (s, 1.6H), 7.48 (s, 0.4H), 7.05 (t, J= 7.8 Hz, 1H), 6.76 (t, J=7.0 Hz, 1H), 6.54 (s, 1H), 3.81 (s, 3H), 1.18 -- 1.00 (m, 3H).

[0323] Example 43A: 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide (I-39A) (Unspecified isomer A, from two possible isomers: (1s,4s) and (1r,4r)) Step 1: A mixture of (1s,4s)-4-amino-1-fluorocyclohexane-1-carboxamide, CF3COOH and (1r,4r)-4-amino-1-fluorocyclohexane-1-carboxamide, CF3COOH A mixture of TBTU (295 mg, 0.92 mmol), 4-(tert-butoxycarbonylamino)-1-fluorocyclohexanecarboxylic acid (200 mg, 0.76 mmol), and DIPEA (0.40 mL, 2.3 mmol) in DMF (4 mL) was stirred at room temperature for 10 minutes, followed by the addition of solid NH4Cl (82 mg, 1.53 mmol). The reaction mixture was shaken at room temperature for 4.7 hours, then stirred overnight at room temperature. Subsequently, 4-methoxybenzylamine (0.10 mL, 0.76 mmol) was added at room temperature, and the reaction mixture was shaken at room temperature for 1 hour. An additional TBTU (246 mg, 0.76 mmol) was added in a single dose at room temperature, and stirring was continued for 7 days at room temperature. The reaction mixture was diluted with Et2O and washed successively with 1 M aqueous HCl, H2O, and finally with saturated aqueous NaHCO3 solution. The organic layer was separated and concentrated under reduced pressure to obtain a crude mixture of (4-fluoro-4-((4-methoxybenzyl)carbamoyl)cyclohexyl)carbamate tert-butyl and (4-carbamoyl-4-fluorocyclohexyl)carbamate tert-butyl, as a pale yellow solid. All materials were stirred in CH₂Cl₂ (6 mL) and TFA (0.94 mL, 12 mmol) at room temperature for 3 hours. The mixture was concentrated under reduced pressure to obtain crude 4-amino-1-fluorocyclohexane-1-carboxamide, CF₃COOH, which was used directly in the next step. MS (ESI) m / z [M+H] + 161.22.

[0324] Step 2: (1s,4s)-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide and (1r,4r)-4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide A suspension of K₂CO₃ (212 mg, 1.53 mmol) and 4-amino-1-fluorocyclohexane-1-carboxamide, CF₃COOH (105 mg, 0.38 mmol) in DMF (3 mL) was shaken at room temperature for 10 min. Then, 6-bromo-8-methyl-2-(methanesulfonyl)pyridino[2,3-d]pyrimidin-7(8H)-one (128 mg, 0.40 mmol) was added in a single addition, followed by shaking for another 10 min. The mixture was then stirred at room temperature for 2 days and 19 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (CH₂Cl₂ solution of SiO₂, CH₂Cl₂ / MeOH / concentrated NH₄OH at 89 / 10 / 1), followed by preparative HPLC (C60 ... 18 Purified with an aqueous solution of MeCN + 0.1% HCO2H2O solution, two isomers ("Isomer 1" and "Isomer 2") were separated: the first eluted "Isomer 1" was a white solid (8.0 mg, yield 5% based on 95% purity), MS (ESI) m / z [M+H2O] + 398.5|400.4; "Isomer 2" is a white solid (10 mg, yield 6% based on 93% purity), MS (ESI) m / z [M+H] + 398.5|400.4.

[0325] Step 3: 4-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide (from "Isomer 2") (I-39A) SMC was prepared using a general method: 4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide ("Isomer 2", 10 mg, 95%, 0.023 mmol), Cs2CO3 (31 mg, 0.095 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (14 mg, 0.038 mmol) and PdCl2dppf*CH2Cl2 (3.9 mg, 4.8 µmol) were heated at 100 °C for 2.8 h in 1,4-dioxane (4 mL) and H2O (2 mL). Purification by rapid chromatography (SiO2, using CH2Cl2 solution with CH2Cl2 / MeOH / concentrated NH4OH at a ratio of 89 / 10 / 1) yielded a white solid 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide (11 mg, 72% yield based on 99% purity). MS (ESI) m / z [M+H + 634.57. 1 H NMR (500 MHz, DMSO-d6) δ 10.37 (s, 1H), 8.66 (s, 1H), 8.46 (s, 1H), 8.08 -- 7.99 (m, 1.6H), 7.90 (brs, 0.4H), 7.75 (s, 1H), 7.53 (s, 1H), 7.37 -- 6.99 (m, 4H), 4.17 --4.04 (m, 1H), 4.04 (s, 3H), 3.89 (s, 3H), 2.28 -- 2.07 (m, 2H), 2.03 -- 1.77(m, 4H), 1.71 -- 1.48 (m, 2H).

[0326] Example 40B: 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide (unspecified isomer B, derived from two possible isomers (I-39B): (1s,4s) and (1r,4r)) SMC was prepared using a general method: 4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide ("Isomer 1", 8 mg, 95%, 0.02 mmol), Cs2CO3 (24 mg, 0.075 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (11 mg, 0.03 mmol), and PdCl2dppf*CH2Cl2 (3.0 mg, 3.7 µmol) in 1,4-dioxane (4 mL) and H2O (2 mL) were heated at 100 °C for 2.6 h in a microwave reactor. Purified by rapid chromatography (CH2Cl2 solution with SiO2, CH2Cl2 / MeOH / concentrated NH4OH at a ratio of 89 / 10 / 1), followed by ultrasonic grinding with Et2O, a white solid 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide (4 mg, yield 32% based on 95% purity) was obtained.

[0327] MS (ESI)m / z [M+H] + 634.42. 1 H NMR(500 MHz, DMSO-d6) δ 8.69 -- 8.59 (m,1H), 8.35 (s, 0.7H), 8.22 (s, 0.3H), 8.03 (s, 1H), 7.93 (d, J = 7.3 Hz,0.7H), 7.82 -- 7.77 (m, 0.3H), 7.72 (s, 1H), 7.56 (s, 1H), 7.35 (s, 1H), 7.22 -- 7.15 (m, 1H), 7.05 -- 6.81 (m, 2H), 4.01 -- 3.89 (m, 1H), 3.85 (s, 3H), 2.09 -- 1.79 (m, 6H), 1.68 -- 1.52 (m, 2H). 19 F NMR (471 MHz, DMSO-d6) δ -73.49, -163.19, -163.31.

[0328] Example 44. (1R,3S)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclopentane-1-carboxamide (cis I-40) Step 1: (1R,3S)-3-amino-N-(4-methoxybenzyl)cyclopentane-1-carboxamide trifluoroacetate (1R,3S)-3-aminocyclopentane-1-carbamate (315 mg, 1.90 mmol) and Boc₂O (436 mg, 2.0 mmol) were suspended in CH₂Cl₂ (10 mL) at room temperature. DIPEA (0.99 mL, 5.7 mmol) was added in a single addition, and the reaction was stirred for 3 days and 21 hours. Subsequently, TBTU (641 mg, 2.0 mmol) was added at room temperature, and the reaction was shaken for 5 minutes at room temperature. Then, 4-methoxybenzylamine (0.27 mL, 2.1 mmol) was added at room temperature. The mixture was stirred overnight at room temperature. The reaction mixture was then diluted with Et₂O, washed (once with 1 M HCl aqueous solution, twice with H₂O), and concentrated under reduced pressure to give crude ((1S,3R)-3-((4-methoxybenzyl)carbamoyl)cyclopentyl)carbamate tert-butyl ester, a white solid. The material was dissolved in CH2Cl2 (7 mL) and stirred with TFA (1.0 mL, 13 mmol) at room temperature for 21 hours. The solvent was removed under reduced pressure, and the residue was dried under high vacuum to give a light orange gel-like substance (1R,3S)-3-amino-N-(4-methoxybenzyl)cyclopentane-1-carboxamide, trifluoroacetic acid MS (ESI) m / z [M+H]. + 249.28.

[0329] Step 2: (1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-(4-methoxybenzyl)cyclopentane-1-carboxamide A suspension of K₂CO₃ (305 mg, 2.21 mmol) and (1R,3S)-3-amino-N-(4-methoxybenzyl)cyclopentane-1-carboxamide and CF₃COOH (200 mg, 0.55 mmol) in DMF (3 mL) was shaken at room temperature for 5 minutes. 6-Bromo-8-methyl-2-(methanesulfonyl)pyridino[2,3-d]pyrimidin-7(8H)-one (184 mg, 0.580 mmol) was added in a single addition, and shaking was continued at room temperature for 1 minute. The mixture was then stirred at room temperature for 20 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by two rounds of rapid chromatography (a CH2Cl2 solution with SiO2, CH2Cl2 / MeOH / concentrated NH4OH at a concentration of 89 / 10 / 1) to give a clear film-like product (1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-(4-methoxybenzyl)cyclopentane-1-carboxamide (55 mg, 21% yield in two steps). MS (ESI) m / z [M+H + 486.35|488.39.

[0330] Step 3: (1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclopentane-1-carboxamide (1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)-N-(4-methoxybenzyl)cyclopentane-1-carboxamide (55 mg, 0.11 mmol) was dissolved in TFA (1.4 mL, 18 mmol) and stirred sequentially at 60 °C for 10 min, at 50 °C for 3 days, at room temperature for 8 days, and finally at 65 °C for 19 h. The reaction mixture was concentrated under reduced pressure to give a gray solid (1R,3S)-3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclopentane-1-carboxamide (41 mg, 94% yield based on 94% purity). This material was used directly in the next step without further purification. MS (ESI)m / z [M+H] + 366.29|368.32.

[0331] Step 4: (1R,3S)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclopentane-1-carboxamide (cis I-40) SMC was prepared according to the general method: cyclopentane-1-carboxamide (41.4 mg, 0.11 mmol), Cs2CO3 (242 mg, 0.74 mmol), H2O (2 mL), Pd(dppf)Cl2*CH2Cl2 (8.7 mg, 11 µmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (54 mg, 0.15 mmol), and 1,4-dioxane (4 mL) were heated at 100 °C for 2.6 h. Purification by rapid chromatography (CH2Cl2 solution with SiO2, CH2Cl2 / MeOH / concentrated NH4OH at a ratio of 89 / 10 / 1) yielded a white solid (1R,3S)-3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)cyclopentane-1-carboxamide (13.0 mg, 20% yield based on 98% purity). MS (ESI) m / z [M+H + 602.52. 1 H NMR(500 MHz, DMSO-d6) δ 10.35 (s,1H), 8.68 -- 8.57 (m, 1H), 8.46 (s, 1H), 8.05 (s, 1.7H), 7.92 (d, J = 6.8 Hz,0.3H), 7.73 (s, 1H), 7.38 (s, 2H), 7.29 -- 7.07 (m, 2H), 6.81 (s, 1H), 4.38 -- 4.24 (m, 1H), 3.85 (s, 3H), 3.56 (s, 3H), 2.75 -- 2.67 (m, 1H), 2.24 --2.04 (m, 1H), 2.00 -- 1.89 (m, 1H), 1.86 -- 1.76 (m, 2H), 1.75 -- 1.54 (m, 2H). 19 F NMR (471 MHz, DMSO-d6) δ -124.26.

[0332] Example 45: 5-Chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((6-oxopiridin-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-41) Step 1: 6-Bromo-8-methyl-2-((6-oxopiperidin-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one A DMF (5 mL) suspension of K₂CO₃ (130 mg, 0.94 mmol), 5-aminopiperidin-2-one (97 mg, 0.85 mmol), and 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (150 mg, 0.47 mmol) was shaken at room temperature for 3.7 h, then stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (a CH₂Cl₂ solution of SiO₂, CH₂Cl₂ / MeOH / concentrated NH₄OH at 89 / 10 / 1) to give a beige solid of 6-bromo-8-methyl-2-((6-oxoperidin-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one (33.0 mg, 19% yield based on 95% purity). MS (ESI)m / z [M+H] + 352.22, 354.17.

[0333] Step 2: 5-Chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((6-oxopiperidin-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-41) Preparation was performed using the general method SMC: 6-bromo-8-methyl-2-((6-oxopiperidin-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one (33 mg, 0.094 mmol), Cs₂CO₃ (153 mg, 0.47 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (61 mg, 0.17 mmol), and PdCl₂dppf*CH₂Cl₂ (11 mg, 0.014 mmol) were heated at 100 °C for 2 hours in 1,4-dioxane (4 mL) and H₂O (2 mL); the solution was obtained by rapid chromatography. (A CH2Cl2 solution with a SiO2 / CH2Cl2 / MeOH / concentrated NH4OH ratio of 8:9 / 10 / 1 ) Purified and then ground with MeOH, the product yielded a beige solid, 5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((6-oxopiperidin-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (10 mg, 17% yield based on 93% purity). MS (ESI) m / z [M+H + 588.4. 1 H NMR(500 MHz, DMSO-d6) δ 10.38 (s, 1H), 8.73 --8.63 (m, 1H), 8.46 (s, 1H), 8.10 -- 8.03 (m, 1H), 7.94 (s, 1H), 7.77 (s, 1H),7.47 (s, 1H), 7.27 (t, J = 7.0 Hz, 1H), 7.23 -- 7.10 (m, 2H), 4.22 (br.s,1H), 3.88 (s, 3H), 3.55 (s, 3H), 3.20 -- 3.06 (m, 1H), 2.40 -- 2.20 (m, 3H),2.11 -- 1.95 (m, 1H), 1.93 -- 1.74 (m, 1H). 19 F NMR (471 MHz, DMSO-d6) δ -123.95.

[0334] Example 46: 3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylic acid tert-butyl ester (I-42) Step 1: 3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylic acid tert-butyl ester A DMF (5 mL) suspension of K2CO3 (174 mg, 1.257 mmol), 1-Boc-3-(amino)azacyclobutane (206 mg, 1.2 mmol), and 6-bromo-8-methyl-2-(methanesulfonyl)pyridino[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.63 mmol) was shaken at room temperature for 10 minutes and then stirred overnight at room temperature for a total of 1 day and 19 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (a CH2Cl2 solution with SiO2, CH2Cl2 / MeOH / concentrated NH4OH at a concentration of 89 / 10 / 1) to give a white solid tert-butyl 3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylate (102 mg, 39%). MS (ESI) m / z [M+H] + 410.45|412.40.

[0335] Step 2: 3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylic acid tert-butyl ester (I-42) Preparation using the general method SMC: tert-butyl 3-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylic acid (50 mg, 0.12 mmol), Cs₂CO₃ (199 mg, 0.61 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (79 mg, 0.22 mmol), and PdCl₂(dppf)*CH₂Cl₂ (15 mg, 0.018 mmol) in 1,4-dioxane (4 mL) and H₂O (2 mL) was performed in a microwave reactor heated to 90 °C for 90 min; the solution was obtained by rapid chromatography (SiO₂, CH₂Cl₂ / MeOH / concentrated NH₄OH). Purified with 89 / 10 / 1 CH2Cl2 solution, the product yielded a white solid tert-butyl 3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylate (39 mg, 50% yield). MS (ESI) m / z [M+H) + 646.5. 1H NMR(500 MHz, DMSO-d6) δ 10.37(brs, 1H), 8.67 (s, 1H), 8.53 -- 8.50 (m, 1H), 8.48 -- 8.39 (m,1H), 8.06 (s,1H), 7.78 (s, 1H), 7.33 -- 6.92 (m, 3H), 4.71 -- 4.52 (m, 1H), 4.28 -- 4.09(m, 2H), 3.88 (s, 3H), 3.90-3.76 (m, 1H), 3.53 (s, 3H), 1.39 (s, 9H). 19 F NMR (471 MHz, DMSO-d6) δ -123.92.

[0336] Example 47: Ethyl 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate (I-43) Step 1: Ethyl 4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate A DMF (5 mL) suspension of K₂CO₃ (174 mg, 1.3 mmol), ethyl 4-aminopiperidin-1-carboxylate (173 mg, 1.0 mmol), and 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.63 mmol) was stirred at room temperature for 2 days. The reaction was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (CH₂Cl₂ solution of SiO₂, CH₂Cl₂ / MeOH / concentrated NH₄OH at 89 / 10 / 1) to give a pale yellow solid of ethyl 4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidin-1-carboxylate (51.0 mg, 19% yield based on 95% purity). MS (ESI)m / z [M+H] + 410.30|412.25.

[0337] Step 2: 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylic acid ethyl ester (I-43) Preparation was performed using a general SMC method: ethyl 4-((6-bromo-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)piperidin-1-carboxylate (51 mg, 0.12 mmol), Cs₂CO₃ (192 mg, 0.59 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (77 mg, 0.21 mmol), and PdCl₂(dppf)*CH₂Cl₂ (14 mg, 0.018 mmol) in 1,4-dioxane (6 mL) and H₂O (3 mL), heated at 90 °C for 2 hours; the solution was analyzed by rapid chromatography (SiO₂, Purification with a CH2Cl2 / MeOH / concentrated NH4OH solution at a concentration of 89 / 10 / 1 yielded a white solid, ethyl 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylate (18 mg, 22% yield based on 95% purity). MS (ESI) m / z [M+H + 646.5. 1 H NMR(500 MHz, DMSO-d6) δ 10.37 (s, 1H), 8.75-- 8.58 (m, 1H), 8.48 (s, 1H), 8.07 (s, 1H), 7.99 (d, J = 6.2 Hz, 0.6H), 7.85(s, 0.4H), 7.75 (s, 1H), 7.30 -- 7.12 (m, 3H), 4.13 -- 3.92 (m, 5H), 3.89 (s,3H), 3.61 -- 3.50 (m, 3H), 3.09 -- 2.78 (m, 2H), 2.01 -- 1.81 (m, 2H), 1.51-- 1.37 (m, 2H), 1.19 (t, J = 7.0 Hz, 3H). 19 F NMR (471 MHz, DMSO-d6) δ -123.96.

[0338] Example 48: N-(3-(2-((1-acetylpiperidin-4-yl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-44) Step 1: 2-((1-acetylpiperidin-4-yl)amino)-6-bromo-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one A suspension of K₂CO₃ (174 mg, 1.257 mmol), 1-acetylpiperidin-4-amine (134 mg, 0.94 mmol), and 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.63 mmol) was stirred at room temperature for 2 days. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO₂, CH₂Cl₂ / / MeOH / concentrated NH₄OH in CH₂Cl₂ solution at 89 / 10 / 1) to give a colorless film-like substance 2-((1-acetylpiperidin-4-yl)amino)-6-bromo-8-methylpyrido[2,3-d]pyrimidin-7(8H)-one (25.0 mg, 10% yield based on 97% purity). MS (ESI) m / z [M+H] + 380.28 | 382.2.

[0339] Step 2: N-(3-(2-((1-acetylpiperidin-4-yl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-44) Preparation was performed using a general SMC method: 2-((1-acetylpiperidin-4-yl)amino)-6-bromo-8-methylpyridino[2,3-d]pyrimidin-7(8H)-one (25 mg, 0.064 mmol), Cs₂CO₃ (104 mg, 0.32 mmol), (3-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (41 mg, 0.11 mmol), and PdCl₂(dppf)*CH₂Cl₂ (7.8 mg, 9.6 µmol) were heated at 90 °C for 2.3 h in 1,4-dioxane (10 mL) and H₂O (5 mL); the precipitate was analyzed by rapid chromatography (SiO₂, Purification with a CH2Cl2 / MeOH / concentrated NH4OH solution at a concentration of 89 / 10 / 1 yielded a white solid N-(3-(2-((1-acetylpiperidin-4-yl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (18 mg, 45% yield based on 99% purity). MS (ESI) m / z [M+H + 616.4. 1 H NMR (500 MHz, DMSO-d6) δ 10.37 (s, 1H), 8.72 -- 8.59 (m, 1H), 8.48 (s, 1H), 8.07 (s, 1H), 8.02 (d, J = 7.2 Hz, 0.6H), 7.87 (d, J =7.0 Hz, 0.4H), 7.76 (s, 1H), 7.31 -- 7.09 (m, 3H), 4.33 -- 4.23 (m, 1H), 4.18 -- 4.01 (m, 1H), 3.89 (s, 3H), 3.83 (d, J = 13.4 Hz, 1H), 3.62 -- 3.51 (m,3H), 3.25 -- 3.09 (m, 1H), 2.85-2.66 (m, 1H), 2.02 (s, 3H), 1.98 -- 1.75 (m, 2H), 1.56 -- 1.44 (m, 1H), 1.41 -- 1.32 (m, 1H). 19 F NMR (471 MHz, DMSO-d6) δ -123.97.

[0340] Example 49: (R)-5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidine-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide ((R) I-45) Step 1: (R)-6-bromo-8-methyl-2-((5-oxopyrrolidone-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one K₂CO₃ (174 mg, 1.26 mmol) and (R)-4-aminopyrrolidone-2-one hydrochloride (129 mg, 0.94 mmol) were shaken in DMF (5 mL) at room temperature for 10 minutes. Then, solid 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.63 mmol) was added in a single batch at room temperature. The reaction mixture was then stirred overnight at room temperature. The reaction was concentrated, and the crude product was used for the next step. MS (ESI) m / z [M+H] + 338.15.

[0341] Step 2: (R)-5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidone-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide ((R) I-45) Preparation was performed using a general method of SMC: (R)-6-bromo-8-methyl-2-((5-oxopyrrolidone-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one (70 mg, 0.21 mmol), Cs2CO3 (337 mg, 1.0 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (134 mg, 0.37 mmol) and PdCl2(dppf)*CH2Cl2 (25 mg, 0.031 mmol) in 1,4-dioxane (4 mL) and H2O (2 mL), heated at 100 °C for 2 h in a microwave reactor; the solution was then analyzed by preparative HPLC (C 18The mixture was purified by a combination of aqueous solution of MeCN and 0.1% HCO2H solution, followed by two rounds of rapid chromatography (SiO2, MeOH in CH2Cl2 solution) and (SiO2, CH2Cl2 / MeOH / concentrated NH4OH in CH2Cl2 solution at a ratio of 89 / 10 / 1) to obtain a white solid (R)-5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidin-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (6.0 mg, 2-step yield 5%). MS (ESI) m / z [M+H + 574.2. 1 H NMR(500 MHz, CD3OD) δ 8.49 (s, 1H),8.21 (d, J = 2.6 Hz, 1H), 7.98 (d, J = 2.6 Hz, 1H), 7.63 (s, 1H), 7.42 --7.34 (m, 1H), 7.11 (dd, J = 10.1 Hz, 3.9 Hz, 1H), 7.05 (t, J = 7.9 Hz, 1H),4.48 (s, 2H), 3.89 (s, 3H), 3.79 -- 3.70 (m, 1H), 3.59 (s, 3H), 3.38 -- 3.27(m, 1H), 2.78 -- 2.61 (m, 1H), 2.44 -- 2.28 (m, 1H). 19 F NMR (471 MHz, CD3OD) delta-127.74.

[0342] Example 50: (S)-5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidine-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide ((S) I-45) Step 1: (S)-6-bromo-8-methyl-2-((5-oxopyrrolidone-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one K₂CO₃ (174 mg, 1.3 mmol), (4S)-4-aminopyrrolidone-2-one (126 mg, 1.3 mmol), and 6-bromo-8-methyl-2-(methanesulfonyl)pyrido[2,3-d]pyrimidin-7(8H)-one (200 mg, 0.63 mmol) were dissolved in DMF (5 mL) and shaken overnight at room temperature. The reaction mixture was concentrated, and the crude product was used for the next step. MS (ESI) m / z [M+H] + 338.2|340.2.

[0343] Step 2: (S)-5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidine-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide ((S) I-45) SMC was prepared using a general method: (S)-6-bromo-8-methyl-2-((5-oxopyrrolidone-3-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one (70 mg, 0.21 mmol), Cs2CO3 (337 mg, 1.03 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (134 mg, 0.37 mmol) and PdCl2dppf·CH2Cl2 (25 mg, 0.031 mmol) in 1,4-dioxane (4 mL) and H2O (2 mL), and heated at 100 °C for 2 hours. The sample was then analyzed by rapid chromatography (SiO2, The CH2Cl2 / MeOH / concentrated NH4OH aqueous solution was prepared as an 8:9 / 10 / 1 CH2Cl2 solution, which was then purified with a MeOH-CH2Cl2 solution. This was followed by preparative HPLC (Cellular HPLC). 18 The solution was purified by a combination of MeCN in H₂O solution + 0.1% HCO₂H and another rapid chromatographic method (SiO₂, CH₂Cl₂ / / MeOH / concentrated NH₄OH aqueous solution in CH₂Cl₂ solution at a concentration of 89 / 10 / 1), yielding (S)-5-chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidin-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a white solid (4 mg, 3% yield in two steps based on 98% purity). MS (ESI) m / z [M+H) + 574.2. 1H NMR (500 MHz, CD3OD) δ 8.49(s, 1H), 8.20 (d, J = 2.6 Hz, 1H), 7.98 (d, J = 2.6 Hz, 1H), 7.63 (s, 1H), 7.36 (dd, J = 10.7, 4.6 Hz, 1H), 7.10 (t, J = 6.3 Hz, 1H), 7.04 (t, J = 7.9Hz, 1H), 4.51 (s, 3H), 3.82 – 3.70 (m, 1H), 3.59 (s, 3H), 3.39 – 3.26 (m,1H), 2.78-2.63 (m, 1H), 2.44 – 2.28 (m, 1H). 19F NM R (471 MHz, CD3OD) δ -127.81.

[0344] Example 51: 5-Chloro-N-(2-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-46) SMC was prepared using a general method: 6-bromo-2-(isopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (11 mg, 0.039 mmol), Cs₂CO₃ (51 mg, 0.15 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (25.2 mg, 0.070 mmol), Pd(dppf)Cl₂·CH₂Cl₂ (4.8 mg, 5.8 µmol) in H₂O (2.5 mL) and 1,4-dioxane (5 mL), heated overnight at 100 °C; purified by rapid chromatography (SiO₂, MeOH in CH₂Cl₂ solution), followed by preparative HPLC (C 18 Purification with MeCN in H₂O solution + 0.1% HCO₂H₂O yielded 5-chloro-N-(2-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a white solid (1.0 mg, 5% yield). MS (ESI) m / z [M+H₂] + 519.1.

[0345] Example 52: 5-Chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-47) Step 1: 6-Bromo-2-(cyclopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one A single addition of c-PrNH2 (0.58 mL, 8.3 mmol) was made to a suspension of 6-bromo-2-(methylsulfinyl)pyrido[2,3-d]pyrimidin-7(8H)-one (120 mg, 0.42 mmol) in i-PrOH (10 mL). The suspension was shaken at room temperature for 1.5 h, then stirred at room temperature for 3 days and 18 h. The solid was collected by filtration and washed with i-PrOH to give 6-bromo-2-(cyclopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one as a white solid (111 mg, 94% yield based on 99% purity). MS (ESI) m / z [M+H] + 281.03|283.06.

[0346] Step 2: 5-Chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-47) SMC was prepared using a general method: 6-bromo-2-(cyclopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (55.5 mg, 0.19 mmol), Cs2CO3 (191 mg, 0.59 mmol), (5-((5-chloro-2-methoxypyridinyl)-3-sulfonamido)-2-fluorophenyl)boronic acid (106 mg, 0.29 mmol) and Pd(dppf)Cl2·CH2Cl2 (24 mg, 0.029 mmol) in 1,4-dioxane (4 mL) and H2O (2 mL) were heated at 100 °C for 2 h in a microwave reactor; purification was performed by rapid chromatography (CH2Cl2 solution of SiO2 and MeOH). A solid precipitated from MeOH / CH₂Cl₂ was collected by filtration and eluted with a small amount of CH₂Cl₂ to give 5-chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide, as a white solid (32 mg, yield 30% based on 95% purity). MS (ESI) m / z [MH - 515.0. 1 H NMR (500 MHz, DMSO-d6) δ 12.15 – 11.92 (m,1H), 10.47 (s, 1H), 8.75 – 8.54 (m, 1H), 8.48 (d, J = 2.6 Hz, 1H), 8.16 (d, J= 2.6 Hz, 1H), 8.06 – 7.79 (m, 1H), 7.74 (s, 1H), 7.23 (dd, J = 6.4, 2.7 Hz,1H), 7.16 (t, J = 9.2 Hz, 1H), 7.11 – 7.06 (m, 1H), 3.98 (s, 3H), 2.96 – 2.81(m, 1H), 0.71 (d, J = 5.4 Hz (2H), 0.56 (s, 2H). 19F NM R (471 MHz, DMSO-d6) δ -119.06.

[0347] Example 53: 5-Chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-48) SMC was prepared using a general method: 6-bromo-2-(cyclopropylamino)pyrido[2,3-d]pyrimidin-7(8H)-one (55.5 mg, 0.195 mmol), Cs2CO3 (191 mg, 0.59 mmol), (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (106 mg, 0.29 mmol) and Pd(dppf)Cl2·CH2Cl2 (24 mg, 0.029 mmol) in 1,4-dioxane (4 mL) and H2O (2 mL) were heated at 100 °C for 2 hours in a microwave reactor; purification was performed by rapid chromatography (CH2Cl2 solution of SiO2 and MeOH). A solid precipitated from MeOH / CH2Cl2 was collected by filtration and washed with a small amount of MeOH to give 5-chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide, as a white solid (6 mg, 6% yield). MS (ESI) m / z [M+H + 517.1. 1 H NMR (500 MHz, DMSO-d6) δ 12.14 – 11.86 (m, 1H), 10.41 (s, 1H), 8.74 – 8.59 (m, 1H), 8.55 (d, J = 2.3 Hz, 1H), 8.13 (d, J = 2.6 Hz, 1H), 8.05– 7.84 (m, 1H), 7.78 (s, 1H), 7.37 – 7.27 (m, 2H), 7.22 (t, J = 7.7 Hz, 1H), 3.96 (s, 3H), 2.96 – 2.85 (m, 1H), 0.76 (d, J = 5.3 Hz, 2H), 0.61 (s, 2H). 19 FNMR (471 MHz, DMSO-d6) δ -124.13.

[0348] Example 54: 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (I-49) A solution of 5-chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylsulfinyl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (122 mg, 0.18 mmol) in i-PrOH (5 mL) was treated with EtNH2 (66-72% aqueous solution, 0.45 mL, 5.5 mmol) and shaken at room temperature for 4.3 h, followed by stirring for 20 h. The reaction mixture was concentrated under reduced pressure to give a white foam, which was dissolved in TFA (2.0 mL, 26 mmol) and heated at 60 °C for 1.3 h, followed by heating at 70 °C for 1.5 h. The reaction mixture was cooled to room temperature. Anisole (0.020 mL, 0.18 mmol) was added, and the reaction mixture was stirred at room temperature for 1 day and 19 h. Volatile substances were removed under high vacuum, and the solid residue was dried under high vacuum. The substance was then reacted with PdCl2 (32.7 mg, 0.184 mmol) in EtOAc (40 mL) and ice-cold AcOH (10 mL) at room temperature under H2 (1 atm) for 3 days and 20 hours, followed by shaking at room temperature under H2 (2 bar) for 2 hours. The reaction mixture was filtered through diatomaceous earth, concentrated under reduced pressure, and dried under high vacuum. The solid residue was treated with trifluoromethanesulfonic acid (2.0 mL, 23 mmol) while stirring in an ice-cold H2O bath. The reaction mixture was vigorously stirred in a cooling bath for 5 minutes, then the cooling bath was removed and stirring continued for 1.3 hours. Ice-cold H2O was then carefully added, forming a gray viscous precipitate, which was collected by filtration, washed with excess H2O, and then purified by rapid chromatography (SiO2, MeOH in CH2Cl2 solution). The substance was further ground with a 5% MeOH solution in DCM, and then ground with Et2O to give 5-chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide, which was an off-white solid (16 mg, 17% yield). 1H NMR (500 MHz, DMSO-d6)δ 12.15 – 11.97 (m, 1H), 10.35 (s, 1H), 8.72 – 8.54 (m, 1H), 8.50 (d, J = 2.3Hz, 1H), 8.06 (d, J = 2.6 Hz, 1H), 7.88 (s, 0.7H), 7.77 (s, 1H), 7.72 (s,0.3H), 7.36 – 7.26 (m, 1H), 7.13 (t, J = 8.6 Hz, 1H), 3.91 (s, 3H), 3.43 –3.36 (m, 2H), 1.15 (t, J = 6.8 Hz, 3H). 19 F NMR (471 MHz, DMSO-d6) δ -113.12, -119.70.

[0349] Example 55: 5-Chloro-N-(2,4-difluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-50) Step 1: 6-Bromo-8-(4-methoxybenzyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one NaH (60%, 0.88 g, 37 mmol) was added fractionally to a DMF (100 mL) solution of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (4.0 g, 14.7 mmol), and the reaction mixture was stirred at room temperature for 1 hour. After 1 hour, 1-(chloromethyl)-4-methoxybenzene (4.83 g, 31 mmol) was added, and the reaction mixture was stirred at room temperature for another 16 hours. The reaction mixture was then diluted with H2O (150 mL) and extracted with EtOAc (3 x 150 mL). The combined organic layers were dried (using anhydrous Na₂SO₄), concentrated, purified by rapid chromatography, and ground with Et₂O to give 6-bromo-8-(4-methoxybenzyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one, a white solid (2.5 g, 43%). LCMS: [M+H] + 392.1.

[0350] Step 2: 6-(3-amino-2,6-difluorophenyl)-8-(4-methoxybenzyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one In a sealed vial, XPhos Pd G2 (108 mg, 0.14 mmol), 6-bromo-8-(4-methoxybenzyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (300 mg, 0.76 mmol), K3PO4 (649 mg, 3.1 mmol), 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)aniline (585 mg, 2.3 mmol), 1,4-dioxane (20 mL), and H2O (2 mL) were degassed with argon and then heated in a microwave reactor at 65 °C for 18 hours. The reaction mixture was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO2, hexane solution of EtOAc) to give 6-(3-amino-2,6-difluorophenyl)-8-(4-methoxybenzyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one as a pale yellow solid (221.0 mg, 66% yield). MS (ESI) m / z [M+H] + 441.37.

[0351] Step 3: 5-Chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylthio)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide Pyridine (0.81 mL, 10 mmol) was added to a CH₂Cl₂ (20 mL) solution of 6-(3-amino-2,6-difluorophenyl)-8-(4-methoxybenzyl)-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (221 mg, 0.50 mmol) and 5-chloro-2-methoxypyridine-3-sulfonyl chloride (182 mg, 0.75 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 0.5 h, and then stirred for 7 days at room temperature. The reactants were washed (once with 1 M HCl aqueous solution, twice with H2O, and once with 1 M HCl aqueous solution). The organic phase was concentrated under reduced pressure, deposited on diatomaceous earth, and purified by rapid chromatography (SiO2, hexane solution of EtOAc) to give 5-chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylthio)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a white solid (245 mg, 73% yield based on 97% purity). MS (ESI) m / z [M+H + 646.39.

[0352] Step 4: 5-Chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylsulfinyl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide A cold (0 °C) CH₂Cl₂ (12 mL) solution of 5-chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylthio)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (245 mg, 0.37 mmol) was added in solid form to mCPBA (102 mg, 0.441 mmol, approximately 75%). The reaction mixture was stirred under cooling for 0.9 h and stored overnight at –20 °C. The mixture was then heated to room temperature, concentrated to dryness under reduced pressure, and the crude product was used for the next step. MS (ESI) m / z [MH] - 660.46.

[0353] Step 5: 5-Chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide To crude 5-chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylsulfinyl)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (122 mg, 0.18 mmol), solid K₂CO₃ (293 mg, 2.1 mmol), MeNH₂·HCl (81 mg, 1.2 mmol), and DMF (4.0 mL) were added. The reaction was carried out at room temperature with shaking for 4.6 h, followed by stirring for 20 h. The mixture was concentrated under reduced pressure and then dried under high vacuum. The solid residue was suspended in H₂O, filtered, washed with H₂O, and dried to give 5-chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide, as a brownish-brown solid (117 mg, quantitative). MS (ESI) m / z [M+H + 629.53.

[0354] Step 6: 5-Chloro-N-(2,4-difluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-50) 5-Chloro-N-(2,4-difluoro-3-(8-(4-methoxybenzyl)-2-(methylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (117 mg, 0.19 mmol) was treated once with trifluoromethanesulfonic acid (1.0 mL, 11 mmol) while cooling in an ice-cold H2O bath. The reaction mixture was vigorously stirred in the cooling bath for 5 min, then the cooling bath was removed and the mixture was stirred for 1.4 h. Ice-cold H2O was carefully added to form a precipitate. The solid was filtered, washed with excess H2O, and analyzed by preparative HPLC (C6000-2000 ppm). 18 Purified using a solution of MeCN in H₂O + 0.1% HCO₂ (MS) and analyzed by MS (ESI) m / z [M+H). + 509.30. 1H NMR (500 MHz, DMSO-d6) δ 12.06 (s, 0.7H), 11.94 (s, 0.3H), 10.28 (s, 1H), 8.60 (s, 0.3H), 8.51 (s, 0.7H), 8.43 (d, J = 2.6 Hz, 1H), 7.99 (d, J= 2.6 Hz, 1H), 7.77 – 7.72 (m, 0.7H), 7.70 (brs, 0.3H), 7.55 (s, 1H), 7.28 –7.19 (m, 1H), 7.07 (t, J = 8.8 Hz, 1H), 3.84 (s, 3H), 2.80 (d, J = 4.6 Hz, 3H).

[0355] B. Biological assays GCN2 Enzyme Assay To identify small-molecule GCN2 inhibitors, a biochemical GCN2 enzymatic assay was outsourced to Eurofins. This assay employed a radiometric method utilizing full-length GST-labeled GCN2 (E556G) produced in insect cells. The kinase concentration was 18.5 nM in Tris buffer containing 300 μM of optimized peptide substrate (RSRSRSRSRSRSRSR), 70 μM ATP (Km=77 μM), and [g-33P]-ATP. The reaction was initiated by adding a Mg / ATP mixture. After incubation at room temperature for 40 min, the reaction was terminated by adding phosphate to a concentration of 0.5%. 10 μL of the reaction solution was then spotted onto a P30 filter membrane, washed four times with 0.425% phosphate for 4 min each time, washed once with methanol, dried, and subjected to scintillation counting. The results are shown in Table 1, where the IC50 of compound (I) is... 50 Report within the following ranges: A: 0.1-100 nM; B: 100-1000 nM; C: 1000-10000 nM; D: >10000 nM.

[0356] Table 1: IC50 of exemplary compounds in this application inhibiting GCN2 50 (nM)

[0357] Cell-based phosphate-eIF2α assay: To confirm target binding in cells, the AlphaLISA assay (Perkin Elmer #TGREIR2S10K) was optimized to monitor eIF2α phosphorylation at serine-51. This event is GCN2-specifically catalyzed and induced by halofopontoxin (a glutamate-prolyl-tRNA synthetase inhibitor), spirochetin (a threonyl-tRNA synthetase inhibitor), or L-asparaginase, which activates GCN2 kinase activity by triggering an amino acid starvation response. SKOV3 or U2OS cells (40,000 cells per well) were pretreated for 1 h with an exemplary GCN2 inhibitor compound of this application (1 nM to 1 µM), stimulated with spirochetin (10 µM) for 1 h, then lysed and analyzed using the AlphaScreen SureFire kit, which uses an antibody-based method to quantify phosphate-eIF2α in HTS mode.

[0358] Tumor cell growth inhibition assay: SKOV3 or OVACR8 cells were seeded at 1000 cells / well in 50 µl of medium (Alpha-MEM containing 10% FBS, 100 mg / ml Normocin (Invivogen), and 50 mg / ml Gentamicin (Invitrogen)) in 384-well plates. The plates were then incubated overnight to allow cell adhesion. ASNase, DMSO, or the test compound was administered to the cells at 16 concentrations (10 µM for high doses to 5 nM for low doses) using an HP D300 digital dispenser. The plates were incubated at 37°C in a humidified 5% CO2 incubator. After 3–5 days, the plates were removed from the incubator and allowed to equilibrate to room temperature. An equal volume of ATPlite assay reagent was then added to each well, and the samples were processed according to the manufacturer's instructions (Perkin Elmer). The luminescence signal was then measured using an Envision plate reader equipped with a US luminescence detector.

[0359] While this application has been described with reference to what is now considered a preferred embodiment, it should be understood that this application is not limited to the disclosed embodiments. Rather, this application is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

[0360] All publications, patents, and patent applications are incorporated herein by reference in their entirety as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated herein by reference in its entirety. Where a term found in this application is defined differently in documents incorporated herein by reference, the definition provided herein shall be used as the definition of that term.

Claims

1. Compounds of Formula I, or pharmaceutically acceptable salts, solvates, and / or prodrugs thereof: (I)， in R 1 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being surrounded by one or two R groups. 8 replace; X 1 Selected from N and CR 9 ; R 2 Selected from H, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 2 Selected from N and CR 10 ; R 3 R 4 and R 5 Independently selected from H, halogens, CN, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; X 3 Selected from N and CR 11 ; R 6 and R 7 Independently selected from H, halogens, CN, C 1-6 Alkyl, C 1-6 Halogenated alkyl, OC 1-6 Alkyl and OC 1-6 Halogenated alkyl groups; Each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 C(O)OR 12 =O, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, wherein all alkyl, alkenyl, ynyl, cycloalkyl, and heterocyclic alkyl groups are optionally selected from one or more halogens, OR 14 NR 14 R 15 and C 1-6 Alkyl substituents; R 9 R 10 and R 11 Independently selected from H, halogens, C 1-6 Alkyl and C 1-6 Halogenated alkyl groups; R 12 Selected from H, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups optionally being selected by one or two from halogen, OH, OC 1-4 Alkyl and OC 1-4 Substituents of fluoroalkyl groups; and R 13 R 14 and R 15 Independently selected from H and C 1-6 Alkyl and C 1-6 Halogenated alkyl groups.

2. The compound according to claim 1, wherein R 1 Selected from C 3-10 cycloalkyl and C3- 10 Heterocyclic alkyl groups, each optionally separated by one or two R 8 replace.

3. The compound according to claim 1, wherein R 1 Selected from H, C 1-4 Alkyl and C 1-4 Fluorinated alkyl groups.

4. The compound according to claim 3, wherein R 1 Selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3 and CH(CH3)3.

5. The compound according to claim 1, wherein R 1 It can be arbitrarily selected by one or two Rs. 8 Replacement C 3-10 Cycloalkyl.

6. The compound according to claim 5, wherein R 1 It can be arbitrarily selected by one or two Rs. 8 Substituted monocyclic C3-8 cycloalkyl groups.

7. The compound according to claim 6, wherein R 1 Selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, each optionally bonded by one or two R... 8 replace.

8. The compound according to claim 7, wherein R 1 Selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl, each of which is randomly diffused by an R 8 replace.

9. The compound according to claim 8, wherein R 1 Selected from cyclobutyl and cyclohexyl, each is converted by an R 8 replace.

10. The compound according to claim 1, wherein R 1 It can be arbitrarily selected by one or two Rs. 8 Replacement C 3-10 Heterocyclic alkyl groups.

11. The compound according to claim 10, wherein R 1 Selected from azircyclopropane, oxacyclopropane, thiohexacyclopropane, azircyclobutane, oxacyclobutane, thiohexacyclobutane, diazircyclobutane, dioxacyclobutane, dithiohexacyclobutane, tetrahydrofuranyl, tetrahydrothiopheneyl, pyrrolidinyl, imidazoalkyl, pyrazolyl, isoxthiolidinyl, thiazoalkyl, isothiazolyl, dioxacyclopentyl, dithiohexacyclopentyl, piperidinyl, tetrahydropyranyl, diazinanyl (e.g., piperazinyl), morpholinyl, thiomorpholinyl, dioxyl, dithiadinyl, azircycloheptyl, oxacycloheptyl, and thiohexacycloheptyl, each optionally separated by one or two R 8 replace.

12. The compound according to claim 11, wherein R 1 Selected from thioheterobutyl, oxoheterobutyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl, azirropropyl, azirrobutyl, pyrrolidinyl, morpholinyl, piperazine, and piperidinyl, each optionally distilled by an R 8 replace.

13. The compound according to claim 12, wherein R 1 Selected from oxetane, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, piperazineyl, and piperidinyl, each optionally distilled by an R 8 replace.

14. The compound according to claim 13, wherein R 1 Selected from oxetane, tetrahydrofuranyl, and tetrahydropyranyl, each optionally converted by an R 8 replace.

15. The compound according to claim 14, wherein R 1 It is selected from oxetane, tetrahydrofuranyl and tetrahydropyranyl, each of which is unsubstituted.

16. The compound according to any one of claims 1 to 15, wherein X 1 Selected from N and CH.

17. The compound according to any one of claims 1 to 16, wherein R 2 Selected from H and CH3.

18. The compound according to any one of claims 1 to 17, wherein X 2 Selected from N and CH.

19. The compound according to any one of claims 1 to 18, wherein R 3 R 4 and R 5 Independently selected from H, Cl, F, Br, CN, C 1-4 Alkyl and C 1-4 Fluorinated alkyl groups.

20. The compound according to claim 19, wherein R 3 R 4 and R 5 It is independently selected from H, Cl, F, CN, CH3 and CF3.

21. The compound according to any one of claims 1 to 18, wherein R 3 and R 5 At least one of them is selected from halogens and CN.

22. The compound according to any one of claims 1 to 21, wherein X 3 Selected from N and CH.

23. The compound according to any one of claims 1 to 22, wherein R 6 and R 7 Independently selected from H, Cl, F, Br, CN, C 1-4 Alkyl, C 1-4 Fluoroalkyl, OC 1-4 Alkyl and OC 1-4 Fluorinated alkyl groups.

24. The compound according to claim 23, wherein R 6 and R 7 It is independently selected from H, Cl, F, CN, CH3, CF3, CH2CH3, OCH3, OCHF2 and OCF3.

25. The compound according to claim 24, wherein R 6 Selected from OCH3 and OCF3, and R 7 Selected from Cl, F, CH3 and CF3.

26. The compound according to claim 25, wherein R 6 Selected from OCH3 and OCF3, and R 7 It is Cl.

27. The compound according to any one of claims 1 to 26, wherein each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 Cl, F, Br, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-6 cycloalkyl and C 3-6 Heterocyclic alkyl groups, wherein all alkyl, cycloalkyl, and heterocyclic alkyl groups are optionally composed of one or more elements selected from Cl, Br, F, OR. 14 NR 14 R 15 and C 1-4 Alkyl substituents, or each R 8 Selected independently from OR 12 C(O)NR 12 R 13 C(O)OR 12 =O,NR 12 R 13 Cl, F, Br, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-6 cycloalkyl and C 3-6 Heterocyclic alkyl groups, wherein all alkyl, cycloalkyl, and heterocyclic alkyl groups are optionally composed of one or more elements selected from Cl, Br, F, OR. 14 NR 14 R 15 and C 1-4 Alkyl substituents.

28. The compound according to claim 27, wherein each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 Cl, F, CH3, CHF2, CH3CH3 and CF3, or each R 8 Selected independently from OR 12 NR 12 R 13 C(O)NR 12 R 13 C(O)OR 12 , =O, Cl, F, CH3, CHF2, CH3CH3 and CF3.

29. The compound according to claim 18, wherein each R 8 It is independently selected from Cl, F, CH3, CHF2, CH3CH3 and CF3.

30. The compound according to claim 29, wherein each R 8 Independently selected from NR 12 R 13 and C(O)NR 12 R 13 , or each R 8 Independently selected from =O and C(O)OR 12 .

31. The compound according to any one of claims 1 to 15, wherein R 9 R 10 and R 11 Independently selected from H, Cl, Br, F, C 1-4 Alkyl and C 1-4 Fluorinated alkyl groups.

32. The compound according to claim 31, wherein R 9 R 10 and R 11 It is H.

33. The compound according to any one of claims 1 to 32, wherein R 12 Selected from H, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-10 cycloalkyl and C 3-10 Heterocyclic alkyl groups, the last four groups of which may be optionally substituted by one or two substituents selected from Cl, F, Br, OH, OCH3 and OCF3.

34. The compound according to claim 33, wherein R 12 Selected from H, CH3 and CF3.

35. The compound according to claim 34, wherein R 12 Selected from H and CH3.

36. The compound according to any one of claims 1 to 35, wherein R 13 Selected from H, C 1-4 Alkyl and C 1-4 Fluorinated alkyl groups.

37. The compound according to claim 36, wherein R 13 Selected from H and CH3.

38. The compound according to any one of claims 1 to 35, wherein R 14 and R 15 It is independently selected from H, CH3 and CF3.

39. The compound according to claim 38, wherein R 14 and R 15 It is independently selected from H and CH3.

40. The compound according to claim 1, wherein the compound is selected from: N-(3-(2-amino-8-isopropyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-ethyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2,4-difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide; trans-5-chloro-N-(2-cyano-3-(2-((4-(dimethylamino)cyclohexyl)amino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2,4-Difluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(cyclopropylamino)-8-methyl-7-oxo-7,8-dihydropteridin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; (R)-5-chloro-N-(4-fluoro-3-(8-methyl-7-oxo-2-((tetrahydrofuran-3-yl)amino)-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(oxetane-3-ylamino)-7-oxo-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-Fluoro-3-(8-methyl-7-oxo-2-((tetrahydro-2H-pyran-4-yl)amino)-7,8-dihydropteridin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; N-(3-(2-amino-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(5-fluoro-4-(8-methyl-2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-fluoro-4-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)pyridin-2-yl)-2-methoxypyridine-3-sulfonamide; trans-4-((6-(5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide; cis-4-((6-(5-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide; trans-4-((6-(5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide; cis-4-((6-(5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide; trans-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclobutane-1-carboxamide; Racemic-(1R,3S)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide; Racemic-(1R,3R)-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide; cis-3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)cyclobutane-1-carboxamide; ((5-chloro-2-methoxypyridin-3-yl)sulfonyl)(2-fluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)aminopotassium; ((5-chloro-2-methoxypyridin-3-yl)sulfonyl)(3-(2-(ethylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)aminopotassium; 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)-1-fluorocyclohexane-1-carboxamide; (1R,3S) -3-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)cyclopentane-1-carboxamide; 5-Chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((6-oxopiperidin-3-yl)amino)-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 3-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-2-yl)amino)azacyclobutane-1-carboxylic acid tert-butyl ester; 4-((6-(3-(((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-8-methyl-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-2-yl)amino)piperidine-1-carboxylic acid ethyl ester; N-(3-(2-((1-acetylpiperidin-4-yl)amino)-8-methyl-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide; (R) -5-Chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidin-3-yl)amino)-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; (S) -5-Chloro-N-(2-fluoro-3-(8-methyl-7-oxo-2-((5-oxopyrrolidin-3-yl)amino)-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(2-(isopropylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(cyclopropylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-7-oxo-7,8-dihydropyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide; and 5-Chloro-N-(2,4-difluoro-3-(2-(methylamino)-7-oxo-7,8-dihydropyridino[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; Or its pharmaceutically acceptable salts, solvates and / or prodrugs.

41. A pharmaceutical composition comprising one or more compounds as described in any one of claims 1 to 40, or pharmaceutically acceptable salts, solvates and / or prodrugs thereof, and a pharmaceutically acceptable carrier.

42. A method for inhibiting generalized regulatory repressor protein 2 (GCN2) in cells of a biological sample or in a patient, comprising administering to said cells an effective amount of one or more compounds as described in any one of claims 1 to 40, or pharmaceutically acceptable salts, prodrugs, and / or solvates thereof.

43. A method of treating a disease, disorder, or symptom that can be treated by inhibiting GCN2, comprising administering to an individual in need a therapeutically effective amount of one or more compounds as described in any one of claims 1 to 40, or pharmaceutically acceptable salts, prodrugs, and / or solvates thereof.

44. The method of claim 43, wherein the disease, disorder, or condition that can be treated by inhibiting GCN2 is a neoplastic condition.

45. The method of claim 43, wherein the disease, disorder, or symptom that can be treated by inhibiting GCN2 is cancer.

46. ​​The method of claim 45, wherein the cancer is selected from one or more of the following: solid tumors, breast cancer, colon cancer, bladder cancer, skin cancer, head and neck cancer, liver cancer, lung cancer, pancreatic cancer, ovarian cancer, prostate cancer, bone cancer, and glioblastoma.

47. The method of claim 43, wherein the disease, disorder or symptom that can be treated by inhibiting GCN2 is a peripheral neuropathy.

48. The method of claim 47, wherein the peripheral neuropathy is a Charcot-Marie-Tuss (CMT) peripheral neuropathy.

49. A method of treating a disease, disorder, or condition that can be treated by inhibiting GCN2, comprising administering to an individual in need a therapeutically effective amount of one or more compounds as described in any one of claims 1 to 40, or pharmaceutically acceptable salts, prodrugs, and / or solvates thereof, in combination with another known agent that can be used to treat a disease, disorder, or condition that can be treated by inhibiting GCN2.

50. The method of claim 49, wherein the disease, disorder, or symptom that can be treated by inhibiting GCN2 is cancer and / or peripheral neuropathy.

51. The method of claim 49, wherein the disease, disorder, or symptom that can be treated by inhibiting GCN2 is cancer, and the one or more compounds of this application are administered or used in combination with one or more additional cancer treatments.

52. The method of claim 51, wherein the one or more additional cancer treatments are chemotherapeutic agents, and the chemotherapeutic agent is cisplatin.

53. The method of claim 51, wherein the one or more additional cancer treatments are chemotherapeutic agents, and the chemotherapeutic agent is L-asparaginase (L-ASNase).

54. The method of claim 51, wherein the one or more additional cancer treatments are small molecule treatments, and the small molecule treatment is a glutaminase inhibitor or an asparagine synthase (ASNS) inhibitor.

55. A method for enhancing the efficacy of one or more cancer treatments for treating cancer, comprising administering an effective amount of one or more compounds as described in any one of claims 1 to 40, or pharmaceutically acceptable salts, prodrugs, and / or solvates thereof, in combination with an effective amount of one or more additional cancer treatments.

56. The method of claim 55, wherein the one or more cancer treatments are chemotherapeutic agents, and the chemotherapeutic agent is cisplatin.

57. The method of claim 55, wherein the one or more cancer treatments are chemotherapeutic agents, and the chemotherapeutic agent is L-asparaginase (L-ASNase).

58. The method of claim 55, wherein the one or more cancer treatments are small molecule treatments, and the small molecule treatment is a glutaminase inhibitor or an asparagine synthase (ASNS) inhibitor.

59. The method of claim 53, wherein the cancer is associated with low asparagine synthase (ASNS) expression, and the one or more additional cancer treatments are L-asparaginase (L-ASNase).

60. The method of claim 53, wherein the cancer is associated with overexpression or dysregulation of asparagine synthase (ASNS), and the one or more additional cancer treatments are one or more asparagine synthase (ASNS) inhibitors and / or L-asparaginase.

61. The method of claim 53, wherein the cancer is associated with low asparaginase synthase (ASNS) expression and low glutaminase expression, and the one or more additional cancer treatments are L-asparaginase (L-ASNase) and / or one or more glutaminase inhibitors.

62. The method of claim 53, wherein the cancer is associated with overexpression or dysregulation of asparaginase (ASNS) and overexpression or dysregulation of glutaminase, and the one or more additional cancer treatments are L-asparaginase (L-ASNase), one or more glutaminase inhibitors and / or one or more asparaginase (ASNS) inhibitors.

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