Triazolopyridopyrimidine and dihydroimidazopyridopyrimidine derivatives as inhibitors of the gcn2 kinase, compositions and uses thereof

Abstract

The present application relates to triazolopyridopyrimidine and dihydroimidazopyridopyrimidine compounds that have activity as inhibitors of the general control nonderepressible 2 (GCN2) kinase, to processes for their preparation, to compositions comprising them, and to their use, for example, in therapy More particularly, the present application relates to compounds useful in the treatment of diseases, disorders or conditions treatable by inhibiting GCN2 kinase such as cancers and neuronal diseases. Formula (I)

Description

TITLE: TRIAZOLOPYRIDOPYRIMIDINE AND DIHYDROIMIDAZOPYRIDOPYRIMIDINE DERIVATIVES AS INHIBITORS OF THE GCN2 KINASE, COMPOSITIONS AND USES THEREOF

[0001] The present application claims the benefit of priority from co-pending U.S. provisional patent application no. 63 / 535,424 filed on August 30, 2023, the contents of which are incorporated herein by reference in the entirety. FIELD

[0002] The present application relates to triazolopyridopyrimidine and dihydroimidazopyridopyrimidine compounds that have activity as inhibitors of the general control nonderepressible 2 (GCN2) kinase, to processes for their preparation, to compositions comprising them, and to their use, for example, in therapy. More particularly, the present application relates to compounds useful in the treatment of diseases, disorders or conditions treatable by inhibiting GCN2 kinase such as cancers and neuronal diseases. BACKGROUND

[0003] The eukaryotic initiation factor 2α (eIF2α) kinase general control nonderepressible 2 (GCN2) drives cellular adaptation to amino acid limitation by activating the integrated stress response (ISR) that induces activating transcription factor 4 (ATF4). The GCN2 kinase mediated cellular adaptations to amino acid limitation occurs through the translational control of gene expression that is primarily executed by eIF2α phosphorylation. Utilizing quantitative phosphoproteomics, Dokladal et al. recently demonstrated that GCN2 targets auxiliary, 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 trimeric eIF2 G protein complex to promote its association with eIF5 which in turn contributes to the inhibition of translation initiation.

[0004] Under distinct stress conditions, cellular ISR is activated by four eukaryotic initiation factor 2 α (eIF2α) kinases: GCN2, protein kinase–like endoplasmic reticulum kinase (PERK), double-stranded RNA-dependent kinase (PRK), and heme-regulated inhibitor (HRK) [Nat Rev Mol Cell Biol 2016, 17:213–226]. These four eIF2α kinases commonly phosphorylate eIF2α at S51, thereby reducing general protein synthesis. However, specific mRNAs with an upstream open reading frame, such as activating transcription factor 4 (ATF4), are selectively translated by delaying translation re-initiation via eIF2α phosphorylation. ATF4 is a key transcription for stress adaptation and subsequently 1 8713541drives transcription of genes 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 often undergo amino acid deprivation, partly because abnormal proliferation increases the need for amino acids to produce proteins, lipids, and nucleic acids, and partly because insufficient and disorganized formation of blood vessels leads to a supply shortage of amino acids. Thus, GCN2 can be important for cancer cell survival and tumor development. Moreover, knockout of GCN2 or ATF4 has been shown to decrease tumor growth in vivo [EMBO. J.2010, 29:2082–2096]. In addition, the GCN2 arm of the ISR has been shown to protect 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 can also be involved in resistance to cancer chemotherapy because sensitization to the antitumor agent L-asparaginase (L-ASNase) is elicited by GCN2 inhibition in cancer cells that express asparagine synthetase (ASNS) at low levels [Proc Natl Acad Sci USA 2018, 115: E7776–E7785]. ASNS catalyzes the biosynthesis of asparagine (Asn) from aspartate and is highly responsive to cellular stress, in particular to intracellular amino acid depletion. ATF4 induces ASNS, [J Biol Chem. 2017;292(49):19952-19958] which in turn sustains Asn levels and suppresses apoptosis while intracellular depletion of Asn induces apoptosis. Thus, ASNS plays a role during 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, ASNS high tumors should be sensitive to inhibition of ASNS activity when combined with L-ASNase and GCN2 inhibition. This combination is a viable strategy to control the growth, proliferation, and migration of cancer cells, eliminate them, or enhance their sensitivity to existing chemotherapy drugs or radiotherapy. It has been suggested that the GCN2- mediated ISR pathway provides promising targets for cancer therapy. Hence, disruption of this pro-oncogenic stress-induced pathway by inhibiting GCN2 is an attractive therapeutic strategy.

[0006] Another important therapeutic area in which ISR activation is implicated is neuronal disease or neuropathy [Science 2021, 373, 1161–1166]. Dominant mutations in ubiquitously expressed transfer RNA (tRNA) synthetase genes cause axonal peripheral neuropathy, accounting for at least six forms of Charcot-Marie-Tooth (CMT) disease. Genetic evidence in mouse and Drosophila models suggests a gain-of-function mechanism. It has been shown that mutant tRNA synthetases activate the integrated stress response (ISR) through the sensor kinase (general control nonderepressible 2). The 2 8713541chronic activation of the ISR contributed to the pathophysiology, and genetic deletion or pharmacological inhibition of GCN2 alleviated the peripheral neuropathy. The activation of GCN2 suggests that the aberrant activity of the mutant tRNA synthetases is still related to translation and that inhibiting GCN2 or the ISR may represent a therapeutic strategy in CMT [Science 2021, 373, 1156–1161].

[0007] Recently, the small molecule inhibitors of GCN2 kinase have been described (WO2021165346, Black Belt TX LTD).

[0008] There remains a need for potent GCN2 kinase inhibitors for the treatment of, for example, cancers and peripheral neuropathy. Also, there is a need for GCN2 kinase inhibitors with selectivity over other kinases. SUMMARY

[0009] The Applicants have developed novel inhibitors of general control nonderepressible 2 (GCN2) kinase.

[0010] Accordingly, the present invention includes a compound of Formula I, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof:wherein R1is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two R8a; ---- is a single or a double bond; X1is selected from CR9and N when ---- is a double bond and X1is selected from CR9R9aand NR9bwhen ---- is a single bond; R2is selected from H, C1-4alkyl and C1-4haloalkyl; X2is selected from N and CR10; R3, R4and R5are independently selected from H, halo, CN, C1-6alkyl and C1-6haloalkyl; X3is selected from N and CR11; 3 8713541R6and R7are independently selected from H, halo, CN, C1-6alkyl, C1-6haloalkyl, OC1-6alkyl and OC1-6haloalkyl; R8is selected from H, C1-4alkyl and C1-4haloalkyl; each R8ais independently selected from OR12, NR12R13, C(O)NR12R13, halo, C1-6alkyl, C1-6haloalkyl, C2-6alkenyl, C2-6alkynyl, C3-10cycloalkyl and C3-10heterocycloalkyl, wherein all alkyl, alkenyl, alkynyl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents selected from halo, OR14, NR14R15and C1-6alkyl; R9, R9a, R10and R11are independently selected from H, halo, C1-6alkyl and C1-6haloalkyl; R9bis selected from H, C1-6alkyl and C1-6haloalkyl; R12is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two substituents selected from halo, OH, OC1-4alkyl and OC1-4fluoroalkyl; and R13, R14and R15are independently selected from H, C1-6alkyl and C1-6haloalkyl.

[0011] The present application also includes a pharmaceutical composition comprising one or more compounds of the application and a pharmaceutically acceptable carrier.

[0012] The present application further includes a method of inhibiting general control nonderepressible 2 (GCN2) in a cell, either in a biological sample or in a patient, comprising administering an effective amount of one or more compounds of the application to the cell.

[0013] The present application also includes a method of treating a disease, disorder or condition that is treatable by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof.

[0014] The present application as also includes a method of treating a disease, disorder or condition that is treatable by inhibiting GCN2 comprising administering a therapeutically effective amount of one or more compounds of the application in combination with another known agent useful for treatment of the disease, disorder or condition treatable by inhibiting GCN2 to a subject in need thereof

[0015] In some embodiments the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer and the one or more compounds of the application are administered or used in combination with one or more additional cancer treatments, such as radiotherapy, chemotherapy (e.g. cisplatin), targeted therapies such as antibody therapies (including anti-PD1 and / or anti- L1 antibodies) and small molecule therapies 4 8713541such as tyrosine-kinase inhibitors therapies, glutaminase inhibitors (e.g., glutaminase-1 (GLS1) inhibitors), and asparagine synthetase (ASNS) inhibitors, immunotherapy, hormonal therapy and anti-angiogenic therapies.

[0016] In some embodiments, the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer and / or a peripheral neuropathy including Charcot-Marie-Tooth (CMT) peripheral neuropathy.

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

[0018] Other features and advantages of the present application will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the application, are given by way of illustration only and the scope of the claims should not be limited by these embodiments but should be given the broadest interpretation consistent with the description as a whole. DETAILED DESCRIPTION I. Definitions

[0019] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present application herein described for which they are suitable as would be understood by a person skilled in the art.

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

[0021] The term “compound of the application” or “compound of the present application” and the like as used herein refers to a compound of Formula I, including pharmaceutically acceptable salts, solvates and / or prodrugs thereof. 5 8713541

[0022] The term “composition of the application” or “composition of the present application” and the like as used herein refers to a composition comprising one or more compounds the application and at least one additional ingredient.

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

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

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

[0026] As used herein, the words “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, unrecited elements or process / method steps.

[0027] As used herein, the word “consisting” and its derivatives, are intended to be close ended terms that specify the presence of stated features, elements, components, groups, integers, and / or steps, and also exclude the presence of other unstated features, elements, components, groups, integers and / or steps.

[0028] The term “consisting essentially of”, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of these features, elements, components, groups, integers, and / or steps.

[0029] Terms of degree such as “substantially”, “about” and “approximately” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not 6 8713541significantly changed. These terms of degree should be construed as including a deviation of at least ^5% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0030] The term “suitable” as used herein means that the selection of the particular compound or conditions would depend on the specific synthetic manipulation to be performed, the identity of the molecule(s) to be transformed and / or the specific use for the compound, but the selection would be well within the skill of a person trained in the art. All process / method steps described herein are to be conducted under conditions sufficient to provide the product shown. A person skilled in the art would understand that all reaction conditions, including, for example, reaction solvent, reaction time, reaction temperature, reaction pressure, reactant ratio and whether or not the reaction should be performed under an anhydrous or inert atmosphere, can be varied to optimize the yield of the desired product and it is within their skill to do so.

[0031] The present application refers to a number of chemical terms and abbreviations used by those skilled in the art. Nevertheless, definitions of selected terms are provided for clarity and consistency.

[0032] The term “protecting group” or “PG” and the like as used herein refers to a chemical moiety which protects or masks a reactive portion of a molecule to prevent side reactions in those reactive portions of the molecule, while manipulating or reacting a different portion of the molecule. After the manipulation or reaction is complete, the protecting group is removed under conditions that do not degrade or decompose the remaining portions of the molecule. The selection of a suitable protecting group can be made by a person skilled in the art. Many conventional protecting groups are known in the art, for example as described in “Protective Groups in Organic Chemistry” McOmie, J.F.W. Ed., Plenum Press, 1973, in Greene, T.W. and Wuts, P.G.M., “Protective Groups in Organic Synthesis”, John Wiley & Sons, 3rdEdition, 1999 and in Kocienski, P. Protecting Groups, 3rd Edition, 2003, Georg Thieme Verlag (The Americas).

[0033] The term “cell” as used herein refers to a single cell or a plurality of cells and includes a cell either in a cell culture or in a subject.

[0034] The term “subject” as used herein includes all members of the animal kingdom including mammals. Thus, the methods and uses of the present application are applicable to both human therapy and veterinary applications. 7 8713541

[0035] The term “pharmaceutically acceptable” means compatible with the treatment of subjects.

[0036] The term “pharmaceutically acceptable carrier” means a non-toxic solvent, dispersant, excipient, adjuvant or other material which is mixed with an active ingredient (for example, one or more compounds of the application) to permit the formation of a pharmaceutical composition, i.e., a dosage form capable of administration to a subject.

[0037] The term “pharmaceutically acceptable salt” means either an acid addition salt or a base addition salt which is suitable for, or compatible with the treatment of subjects.

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

[0039] A base addition salt suitable for, or compatible with, the treatment of subjects is any non-toxic organic or inorganic base addition salt of any acidic compound.

[0040] The term “prodrug” as used herein means a compound, or salt and / or solvate of a compound, that, after administration, is converted into an active drug.

[0041] The term “solvate” as used herein means a compound, or a salt or prodrug of a compound, wherein molecules of a suitable solvent are incorporated in the crystal lattice.

[0042] The term “inert organic solvent” as used herein refers to a solvent that is generally considered as non-reactive with the functional groups that are present in the compounds to be combined together in any given reaction so that it does not interfere with or inhibit the desired synthetic transformation. Organic solvents are typically non-polar and dissolve compounds that are non-soluble in aqueous solutions.

[0043] The term “alkyl” as used herein, whether it is used alone or as part of another group, means straight or branched chain, saturated alkyl groups. The number of carbon atoms that are possible in the referenced alkyl group are indicated by the prefix “Cn1-n2”. For example, the term C1-10alkyl means an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.

[0044] The terms “halo” or “halogen” as used herein, whether it is used alone or as part of another group, refers to a halogen atom and includes fluoro, chloro, bromo and iodo.

[0045] The term “haloalkyl” as used herein refers to an alkyl group as defined above in which one or more of the available hydrogen atoms have been replaced with a halogen atom. Thus, for example, “C1-6haloalkyl” (or “C1-C6haloalkyl”) refers to a C1to C6linear or branched alkyl group as defined above with or more halogen substituents. 8 8713541

[0046] The term “fluoroalkyl” as used herein refers to an haloalkyl group as defined above wherein the halogen atom is fluoro.

[0047] The term "chloroalkyl" as used herein refers to an haloalkyl group as defined above wherein the halogen atom is chloro.

[0048] The term “available”, as in “available hydrogen atoms” or “available atoms” refers to atoms that would be known to a person skilled in the art to be capable of replacement by another atom or group.

[0049] The term “alkenyl” as used herein, whether it is used alone or as part of another group, means straight or branched chain, unsaturated alkyl groups containing at least one double bond. The number of carbon atoms that are possible in the referenced alkylene group are indicated by the prefix “Cn1-n2”. For example, the term C2-6alkenyl means an alkenyl group having 2, 3, 4, 5 or 6 carbon atoms and at least one double bond.

[0050] The term “alkynyl” as used herein, whether it is used alone or as part of another group, means straight or branched chain, unsaturated alkynyl groups containing at least one triple bond. The number of carbon atoms that are possible in the referenced alkyl group are indicated by the prefix “Cn1-n2”. For example, the term C2-6alkynyl means an alkynyl group having 2, 3, 4, 5 or 6 carbon atoms.

[0051] The term “cycloalkyl,” as used herein, whether it is used alone or as part of another group, means a saturated carbocyclic group containing from 3 to 20 carbon atoms and one or more rings. The number of carbon atoms that are possible in the referenced cycloalkyl group are indicated by the numerical prefix “Cn1-n2”. For example, the term C3-10cycloalkyl means a cycloalkyl group having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.

[0052] The term “heterocycloalkyl” as used herein, whether it is used alone or as part of another group, refers to cyclic groups containing at least one non-aromatic ring containing from 3 to 10 atoms in which one or more of the atoms are a heteromoiety selected from O, S, S(O), SO2, N, NH and N(C1-6alkyl), and the remaining atoms are C. Heterocycloalkyl groups are either saturated or unsaturated (i.e. contain one or more double bonds). When a heterocycloalkyl group contains the prefix Cn1-n2 this prefix indicates the number of carbon atoms in the corresponding carbocyclic group, in which one or more, suitably 1 to 5, of the ring atoms is replaced with a heteroatom as defined above. Heterocycloalkyl groups are optionally benzofused. 9 8713541

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

[0054] The term “benzofused” as used herein refers to a polycyclic group in which a benzene ring is fused with another ring.

[0055] A first ring being “fused” with a second ring means the first ring and the second ring share two adjacent atoms there between.

[0056] A first ring being “bridged” with a second ring means the first ring and the second ring share two non-adjacent atoms there between.

[0057] A first ring being “spirofused” with a second ring means the first ring and the second ring share one atom there between.

[0058] The term “optionally substituted” as used herein means that the referenced group is unsubstituted or substituted.

[0059] The term “substituted” as used herein means that the referenced atom contains at least one substituent group other that a hydrogen atom.

[0060] When a group is substituted with one or more substituents, it understood that the selection of those substituents is independent of each other. That is, the one or more substituents may be the same or different.

[0061] The symbol when drawn perpendicularly across a bond indicates a point of covalent attachment of a chemical group.

[0062] The term “LCMS” as used herein refers to liquid chromatography-mass spectrometry.

[0063] The term “NMR” as used herein refers to nuclear magnetic resonance.

[0064] The term “aq.” as used herein refers to aqueous.

[0065] The term “N” as used herein, for example in “4N”, refers to the unit symbol of normality to denote "eq / L".

[0066] The term “M” as used herein, for example in 4M, refers to the unit symbol of molarity to denote "moles / L".

[0067] The term “DIPEA” as used herein refers to N,N-diisopropyl ethylamine.

[0068] The term “DMF” as used herein to dimethylformamide. 10 8713541

[0069] The term “THF” as used herein refers to tetrahydrofuran.

[0070] The term “DMSO” as used herein refers to dimethylsulfoxide.

[0071] The term “EtOAc” as used herein refers to ethyl acetate.

[0072] The term “MeOH” as used herein refers to methanol.

[0073] The term “EtOH” as used herein refers to ethanol.

[0074] The term “MeCN” or “ACN” as used herein refers to acetonitrile.

[0075] The term “HCl” as used herein refers to hydrochloric acid.

[0076] The term “TFA” as used herein refers to trifluoroacetic acid.

[0077] The term “Hex” as used herein refers to hexanes.

[0078] The term “PBS” as used herein refers to phosphate-based buffer.

[0079] The term “IPA” as used herein refers to isopropyl alcohol.

[0080] The term “dppf” as used herein refers to 1,1'-bis(diphenylphosphino)ferrocene.

[0081] The term “RT” or “rt” as used herein refers to room temperature.

[0082] The term “HPLC” as used herein refers to high-performance liquid chromatography.

[0083] The term “PPA” as used herein refers to polyphosphoric acid.

[0084] The term “TEA” or “Et3N” as used herein refer to triethylamine.

[0085] The term “EDTA” as used herein refers to ethylenediaminetetraacetic acid.

[0086] The term “ATP” as used herein refers to adenosine triphosphate.

[0087] The term “FBS” as used herein refers to fetal bovine serum.

[0088] The term “MEM” as used herein refers to Minimum Essential Medium.

[0089] The term “treating” or “treatment” as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of a disease, disorder or condition, stabilized (i.e. not worsening) state of a disease, disorder or condition, preventing spread of a disease, disorder or condition, delay or slowing of a disease, disorder or condition progression, amelioration or palliation of a disease, disorder or condition state, diminishment of the reoccurrence of a disease, disorder or condition, and 11 8713541remission (whether partial or total), whether detectable or undetectable. “Treating” and “treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. “Treating” and “treatment” as used herein also include prophylactic treatment.

[0090] “Palliating” a disease, disorder or condition means that the extent and / or undesirable clinical manifestations of the disease, disorder or condition are lessened and / or time course of the progression is slowed or lengthened, as compared to not treating the disease, disorder or condition.

[0091] The term “prevention” or “prophylaxis”, or synonym thereto, as used herein refers to a reduction in the risk or probability of a subject becoming afflicted with a disease, disorder or condition treatable by inhibition of GCN2 or manifesting a symptom associated with a disease, disorder or condition treatable by inhibition of GCN2.

[0092] As used herein, the term “effective amount” or “therapeutically effective amount” means an amount of a compound, or one or more compounds, of the application that is effective, at dosages and for periods of time necessary to achieve the desired result.

[0093] The term “disease, disorder or condition treatable by inhibiting GCN2” means that the disease, disorder or condition to be treated is affected by, modulated by and / or has some biological basis, either direct or indirect, that includes GCN2 activity, in particular, increased GCN2 activity. These diseases respond favourably when GCN2 activity associated with the disease, disorder or condition is inhibited by one or more of the compounds or compositions of the application.

[0094] The expression “inhibiting GCN2” as used herein refers to inhibiting, blocking and / or disrupting the kinase activity or function of GCN2 in a cell. The inhibiting, blocking and / or disrupting causes a therapeutic effect in the cell.

[0095] By “inhibiting, blocking and / or disrupting” it is meant any detectable inhibition, block and / or disruption in the presence of a compound compared to otherwise the same conditions, except for in the absence in the compound.

[0096] The term “GCN2” as used herein refers to General Control Nonderepressible 2, or any functional mutant or analogous forms thereof.

[0097] The expression “low asparagine synthetase (ASNS) expression” as used herein means any detectable decrease or reduction in the level of asparagine synthetase (ASNS) in a cancer cell, under otherwise the same except in a healthy cell. 12 8713541

[0098] The expression “asparagine synthetase (ASNS) overexpression or dysregulation” as used herein means any detectable increase in the level of asparagine synthetase (ASNS) in a cancer cell, under otherwise the same conditions, except in a healthy cell.

[0099] The expression “low glutaminase expression” as used herein means any detectable decrease or reduction in the level of glutaminase (e.g., GLS1) in a cancer cell, under otherwise the same conditions, except in a healthy cell.

[0100] The expression “glutaminase overexpression or dysregulation” as used herein means any detectable increase in the level of glutaminase (e.g., GLS1) in a cancer cell, under otherwise the same conditions, except in a healthy cell.

[0101] The term “GLS1” as used herein refers to the “kidney type” glutaminase or any functional mutant or analogous forms thereof.

[0102] The term “administered” as used herein means administration of a therapeutically effective amount of a compound, or one or more compounds, or a composition of the application to a cell or a subject.

[0103] The term “neoplastic disorder” as used herein refers to a disease, disorder or condition characterized by cells that have the capacity for autonomous growth or replication, e.g., an abnormal state or condition characterized by proliferative cell growth. The term “neoplasm” as used herein refers to a mass of tissue resulting from the abnormal growth and / or division of cells in a subject having a neoplastic disorder. Neoplasms can be benign (such as uterine fibroids and melanocytic nevi), potentially malignant (such as carcinoma in situ) or malignant (i.e., cancer).

[0104] The term “fibrosis” as used herein refers to a disease, disorder or condition the thickening and scarring of connective tissue, usually as a result of injury. II. Compounds of the Application

[0105] Triazolopyridopyrimidine and dihydroimidazopyridopyrimidine compounds of the present application were prepared and found to inhibit kinase general control nonderepressible 2 (GCN2).

[0106] Accordingly, the present application includes a compound of Formula I, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof: 13 8713541I) wherein R1is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two R8a; ---- is a single or a double bond; X1is selected from CR9and N when ---- is a double bond and X1is selected from CR9R9aand NR9bwhen ---- is a single bond; R2is selected from H, C1-4alkyl and C1-4haloalkyl; X2is selected from N and CR10; R3, R4and R5are independently selected from H, halo, CN, C1-6alkyl and C1-6haloalkyl; X3is selected from N and CR11; R6and R7are independently selected from H, halo, CN, C1-6alkyl, C1-6haloalkyl, OC1-6alkyl and OC1-6haloalkyl; R8is selected from H, C1-4alkyl and C1-4haloalkyl; each R8ais independently selected from OR12, NR12R13, C(O)NR12R13, halo, C1-6alkyl, C1- 6haloalkyl, C2-6alkenyl, C2-6alkynyl, C3-10cycloalkyl and C3-10heterocycloalkyl, wherein all alkyl, alkenyl, alkynyl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents selected from halo, OR14, NR14R15and C1-6alkyl; R9, R9a, R10and R11are independently selected from H, halo, C1-6alkyl and C1-6haloalkyl; R9bis selected from H, C1-6alkyl and C1-6haloalkyl; R12is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two substituents selected from halo, OH, OC1-4alkyl and OC1-4fluoroalkyl; and R13, R14and R15are independently selected from H, C1-6alkyl and C1-6haloalkyl. 14 8713541

[0107] In some embodiments, R1is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one R8a. In some embodiments, R1is selected from C3-10cycloalkyl and C3-10heteroycloalkyl, each of which is optionally substituted with one or two R8a.

[0108] In some embodiments, R1is selected from H, C1-6alkyl and C1-6haloalkyl. In some embodiments, R1is selected from H, C1-4alkyl and C1-4haloalkyl. In some embodiments, R1is selected from H, C1-4alkyl and C1-4fluoroalkyl. In some embodiments, R1is selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3 and CH(CH3)3. In some embodiments, R1is selected from H, CH3, CH2CH3 and CH(CH3)2.

[0109] In some embodiments, R1is C3-10cycloalkyl optionally substituted with one or two R8a. In some embodiments, R1is a monocyclic C3-10cycloalkyl or a bicyclic C5- 10cycloalkyl, each of which is optionally substituted with one or two R8a. In some embodiments, R1is monocyclic C3-8cycloalkyl optionally substituted with one or two R8a. In some embodiments, the monocyclic C3-8cycloalkyl in R1is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, each of which is optionally substituted with one or two R8a. In some embodiments, R1is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, each of which are unsubstituted. In some embodiments, R1is cyclopropyl. In some embodiments, R1is selected from cyclobutyl, cyclopentyl and cyclohexyl, each of which is substituted with one or two R8a. In some embodiments, R1is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, each of which is optionally substituted with one R8a. In some embodiments, R1is cyclohexyl substituted with one or two R8a. In some embodiments, R1is selected from cyclobutyl and cyclohexyl, each of which is substituted with one R8a. In some embodiments, R1is cyclobutyl substituted with one R8a. In some embodiments, R1is cyclohexyl substituted with one R8a.

[0110] In some embodiments, R1is a spirofused C5-10cycloalkyl or a bridged C5- 10cycloalkyl each of which is optionally substituted with one or two R8a. In some embodiments, the spirofused C5-10cycloalkyl 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 of which is optionally substituted with one or two R8a. In some embodiment, the spirofused C6-15 871354110cycloalkyl is spiro[3.3]heptane substituted optionally with one or two R8a. In some embodiments, the spirofused C5-10cycloalkyl .

[0111] In some embodiments, R1is a optionally substituted with one or two R8a. In some embodiments, the bridged C5-10cycloalkyl is selected from a bicyclopentanyl, a bicycloheptanyl and a bicyclooctanyl each of which optionally substituted with one or two R8a. In some embodiments, the bridged C5-10cycloalkyl is selected from a bicyclopentanyl, a bicycloheptanyl and a bicyclooctanyl each of which is optionally substituted with one R8a. In some embodiments, the bridged C5-10cycloalkyl is selected from.

[0112] In some embodiments, R1is C3-10heterocycloalkyl optionally substituted with one or two R8a. In some embodiments, R1is C3-10heterocycloalkyl optionally substituted with one R8a. In some embodiments, R1is C3-6heterocycloalkyl optionally substituted with one R8a. In some embodiments, R1is selected from aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, diazetidinyl, dioxetanyl, dithietanyl, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, isoxthiolidinyl, thiazolidinyl, isothiazolidinyl, dioxolanyl, dithiolanyl, piperidinyl, tetrahydropyranyl, diazinanyl (e.g., piperazinyl), morpholinyl, thiomorpholinyl, dioxanyl, dithianyl, azepanyl oxepanyl and thiepanyl, each of which is optionally substituted with one or two R8a. In some embodiments, R1is selected from thietanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, thiomorpholinyl, aziridinyl, azetidinyl, pyrrolidinyl, morpholinyl, piperazinyl and piperidinyl, each of which is optionally substituted with one R8a. In some embodiments, R1is selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, piperazinyl, and piperidinyl, each of which is optionally substituted with one R8a. In some embodiments, R1is selected from oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is optionally substituted with one R8a. In some embodiments, R1is selected from oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is unsubstituted.

[0113] In some embodiments, R2is selected from is selected from H, C1-4alkyl and C1-4fluoroalkyl. In some embodiments, R2is selected from H, CH3, CF3, CHF2, CH2CH3, 16 8713541CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3and CH(CH3)3. In some embodiments, R2is selected from H, CH3, CH2CH3and CH(CH3)2. In some embodiments, R2is selected from H and CH3. In some embodiments, R2is H.

[0114] In some embodiments, X1is selected from CR9and N when ---- is a double bond and X1is CR9R9awhen ---- is a single bond. In some embodiments, X1is N when ---- is a double bond. In some embodiments, X1is CR9when ---- is a double bond. In some embodiments X1is CR9R9awhen ---- is a single bond.

[0115] In some embodiments X2is selected from N and CH. In some embodiments, X1is N. In some embodiments X2is CH.

[0116] In some embodiments, R3, R4and R5are not all H. In some embodiments, R3, R4and R5are independently selected from H, halo, CN, C1-4alkyl and C1-4haloalkyl. In some embodiments, at least one of R3, R4and R5is selected from halo, CN and C1- 4haloalkyl. In some embodiments, at least one of R3and R5is selected from halo, CN and C1-4haloalkyl. In some embodiments, at least one of R3, R4and R5is selected from halo and CN. In some embodiments, at least one of R3and R5is selected from halo and CN. In some embodiments, at least one of R3, R4and R5is halo. In some embodiments, at least one of R3and R5is halo In some embodiments, R3, R4and R5are independently selected from H, Cl, F, Br, CN, C1-4alkyl and C1-4fluoroalkyl..In some embodiments, at least one of R3, R4and R5is selected from F, Cl, Br, CN and C1-4fluoroalkyl. In some embodiments, at least one of R3and R5is selected from F, Cl, Br, CN and C1-4fluoroalkyl. In some embodiments, R3, R4and R5are independently selected from H, Cl, F, CN, CH3 and CF3. In some embodiments, R3, R4and R5are independently selected from H, F and CN. In some embodiments, at least one of R3, R4and R5is selected from Cl, F, CN and CF3. In some embodiments, at least one of R3, R4and R5is selected from Cl, F and CN. In some embodiments, at least one of R3and R5is selected from Cl, F and CN. In some embodiments, at least one of R3, R4and R5is selected from F and CN. In some embodiments, at least one of R3and R5is selected from F and CN. In some embodiments, at least one of R3and R5is selected from F and CN and R4is H. In some embodiments, R3is CN, R4is H and R5is H. In some embodiments, R3is H, R4is H and R5is CN. In some embodiments, R3and R5are independently selected from H, F and CN and R4is H. In some embodiments, R3and R5are independently selected from H and F and R4is H. In some embodiments, at least one of R3and R5is F. In some embodiments, at least one of R3and R5is F and R4is H. In some embodiments, R3and R5are both F and R4is H. In 17 8713541some embodiments, R3is F, R4is H and R5is F. In some embodiments, R3is F, R4is H and R5is H. In some embodiments, R3is H, R4is H and R5is F.

[0117] In some embodiments, X3is selected from N and CH. In some embodiments, X3is CH.

[0118] In some embodiments, R6and R7are independently selected from H, Cl, F, Br, CN, C1-4alkyl, C1-4fluoroalkyl, OC1-4alkyl and OC1-4fluoroalkyl. In some embodiments, R6and R7are independently selected from H, Cl, F, CN, CH3, CHF2, CF3, CH2CH3, CH2CH2F, OCH3, OCHF2and OCF3. In some embodiments, R6and R7are independently selected from Cl, F, CH3, CHF2, CF3, CH2CH3, CH2CH2F, OCH3, OCHF2 and OCF3. In some embodiments, R6is selected from OCH3 and OCF3 and R7is selected from Cl, F, CH3, and CF3. In some embodiments, R6is selected from OCH3 and OCF3 and R7is Cl. In some embodiments, R6is OCH3 and R7is Cl.

[0119] In some embodiments, R8is selected from is selected from H, C1-4alkyl and C1-4fluoroalkyl. In some embodiments, R8is selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3 and CH(CH3)3. In some embodiments, R8is selected from H, CH3, CH2CH3 and CH(CH3)2. In some embodiments, R8is selected from H and CH3. In some embodiments, R8is H.

[0120] In some embodiments, each R8ais independently selected from OR12, C(O)NR12R13, NR12R13, halo, C1-4alkyl, C1-4haloalkyl, C2-4alkenyl, C2-4alkynyl, C3-10cycloalkyl and C3-10heterocycloalkyl, wherein all alkyl, alkenyl, alkynyl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents selected from halo, OR14, NR14R15and C1-4alkyl. In some embodiments, each R8ais independently selected from OR12, C(O)NR12R13, NR12R13, Cl, F, Br, C1-4alkyl, C1-4fluoroalkyl, C3-6cycloalkyl and C3- 6heterocycloalkyl, wherein all alkyl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents selected from Cl, Br, F, OR14, NR14R15and C1- 4alkyl. In some embodiments, each R8ais independently selected from OR12, NR12R13, C(O)NR12R13, Cl, F, CHF2, CH3, CH3CH3 and CF3. In some embodiments, each R8ais independently selected from Cl, F, CH3, CHF2, CH3CH3 and CF3. In some embodiments, each R8ais independently selected from CH3, CHF2, CH3CH3 and CF3. In some embodiments, each R8ais independently selected from OR12, NR12R13and C(O)NR12R13. In some embodiments, each R8ais independently selected from NR12R13and C(O)NR12R13.

[0121] In some embodiments, R9, R9a, R10and R11are independently selected from H, Cl, Br, F, C1-4alkyl and C1-4fluoroalkyl. embodiments, R9, R9a, R10and R11are 18 8713541independently selected from H, Cl, F, CH3, CF3, CHF2, CH2CH3, CH2CH2F CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3and CH(CH3)3. In some embodiments, R9, R9a, R10and R11are independently selected from H, Cl, F, CH3and CF3. In some embodiments, R9, R9a, R10and R11are independently selected from H and F. In some embodiments, R9, R9a, R10and R11are H.

[0122] In some embodiments, R9bis selected from H, C1-4alkyl and C1-4fluoroalkyl. In some embodiments, R9bis selected from HCH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3and CH(CH3)3. In some embodiments, R9bis selected from H, CH3 and CF3. In some embodiments, R9bis selected from H and CH3.

[0123] In some embodiments, R12is selected from H, C1-4alkyl, C1-4fluoroalkyl, C3- 10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two substituents selected from Cl, F, Br, OH, OCH3 and OCF3. In some embodiments, R12is selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3 and CH(CH3)3. In some embodiments, R12is selected from H, CH3 and CF3. In some embodiments, R12is selected from H and CH3. In some embodiments, R12is CH3.

[0124] In some embodiments, R13is selected from H, C1-4alkyl and C1-4fluoroalkyl. In some embodiments, R13is selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3 and CH(CH3)3. In some embodiments, R13is selected from H, CH3 and CF3. In some embodiments, R13is selected from H and CH3.

[0125] In some embodiments, R14and R15are independently selected from H, C1- 4alkyl and C1-4fluoroalkyl. In some embodiments, R14and R15are independently selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3 and CH(CH3)3. In some embodiments, R14and R15are independently selected from H, CH3 and CF3. In some embodiments, R14and R15are independently selected from H and CH3.

[0126] In some embodiments, the compounds of Formula I are selected from: Compound I.D. Structure CHEMICAL NAME 5-Chloro-N-(4-fluoro-3-(2- (methylamino)- I-1 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 19 8713541Compound I.D. Structure CHEMICAL NAME trans-5-Chloro-N-(3-(2- (((1R,4R)-4- (dimethylamino)cyclohexyl) trans I-2. amino)- HCO2H [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2- methoxypyridine-3- sulfonamide formic acid salt; N-(3-(2-Amino- [1,2,4]triazolo[4',3':1,6]pyrid I-3 o[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-5-chloro-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(4-fluoro-3-(2- (isopropylamino)- I-4 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(2-fluoro-3-(2- (methylamino)- I-5 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 20 8713541Compound I.D. Structure CHEMICAL NAME 5-Chloro-N-(2,4-difluoro-3- (2-(methylamino)- I-6 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(3-(2- (ethylamino)- I-7 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(2,4-difluoro-3- (2-(isopropylamino)- I-8 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(3-(2- (ethylamino)- I-9 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(3-(2- (cyclopropylamino)- I-10 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(4-fluoro-3-(2- (oxetanylamino)- I-11 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 21 8713541Compound I.D. Structure CHEMICAL NAME 5-Chloro-N-(3-(2- (ethylamino)imidazo[1',2':1, I-12 6]pyrido[2,3-d]pyrimidin-6- yl)-4-fluorophenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(3-(2- (ethylamino)-8,9- I-13 dihydroimidazo[1',2':1,6]pyri do[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(2-fluoro-3-(2- (methylthio)- I-14 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(4-fluoro-3-(9- methyl-2-(methylamino)- I-15 [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; 5-Chloro-N-(2-fluoro-3-(9- methyl-2-(methylamino)- I-16 [1,2,4]triazolo[4’,3’:1,6]pyrid o[2,3-d]pyrimidin-6- yl)phenyl)-2- methoxypyridine-3- sulfonamide; trans-4-((6-(5-((5-Chloro-2- trans I-17 methoxypyridine)-3- sulfonamido)-2- fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-2- yl)amino)-N- 22 8713541Compound I.D. Structure CHEMICAL NAME methylcyclohexane-1- carboxamide; trans-4-((6-(3-((5-Chloro-2- methoxypyridine)-3- sulfonamido)-2- trans I-18 fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-2- yl)amino)-N- and methylcyclohexane-1- carboxamide; trans-4-((6-(3-((5-Chloro-2- methoxypyridine)-3- sulfonamido)-2- trans I-19 fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrid o[2,3-d]pyrimidin-2- yl)amino)cyclohexane-1- carboxamide; I-1 K ((5-Chloro-2-methoxypyridin- 3-yl)sulfonyl)(4-fluoro-3-(2- (methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[ 2,3-d]pyrimidin-6- yl)phenyl)amide, Potassium; I-20 5-Chloro-N-(3-(2- (ethylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[ 2,3-d]pyrimidin-6-yl)-2- fluorophenyl)-2- methoxypyridine-3- sulfonamide; I-21 5-Chloro-N-(3-(2- (cyclopropylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[ 2,3-d]pyrimidin-6-yl)-2- fluorophenyl)-2- methoxypyridine-3- sulfonamide; 23 8713541Compound I.D. Structure CHEMICAL NAME I-22 5-Chloro-N-(2-fluoro-3-(2- (isopropylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[ 2,3-d]pyrimidin-6-yl)phenyl)- 2-methoxypyridine-3- sulfonamide; I-23 5-Chloro-N-(3-(2- (cyclopropylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[ 2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-2- methoxypyridine-3- sulfonamide ; and I-24 N-(3-(2-Amino- [1,2,4]triazolo[4',3':1,6]pyrido[ 2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-5-chloro-2- methoxypyridine-3- sulfonamide, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof.

[0127] In some embodiments the pharmaceutically acceptable salt is an acid addition salt or a base addition salt. The selection of a suitable salt may be made by a person skilled in the art (see, for example, S. M. Berge, et aI., "Pharmaceutical Salts," J. Pharm. Sci.1977, 66, 1-19).

[0128] An acid addition salt suitable for, or compatible with, the treatment of subjects is any non-toxic organic or inorganic acid addition salt of any basic compound. Basic compounds that form an acid addition salt include, for example, compounds comprising an amine group. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric, nitric and phosphoric acids, as well as acidic metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids which form suitable salts include mono-, di- and tricarboxylic acids. Illustrative of such organic acids are, for example, acetic, formic, trifluoroacetic, propionic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, mandelic, salicylic, 2- phenoxybenzoic, p-toluenesulfonic acid and other sulfonic acids such as methanesulfonic acid, ethanesulfonic acid and 2-hydroxyethanesulfonic acid. In an embodiment, the mono- 24 8713541or di-acid salts are formed, and such salts exist in either a hydrated, solvated or substantially anhydrous form. In general, acid addition salts are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection criteria for the appropriate salt will be known to one skilled in the art. Other non-pharmaceutically acceptable salts such as but not limited to oxalates may be used, for example in the isolation of compounds of the application for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt. In some embodiments, the acid addition salt is a hydrochloric or formic acid addition salt.

[0129] A base addition salt suitable for, or compatible with, the treatment of subjects is any non-toxic organic or inorganic base addition salt of any acidic compound. Acidic compounds that form a basic addition salt include, for example, compounds comprising a carboxylic acid group. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium or barium hydroxide as well as ammonia. Illustrative organic bases which form suitable salts include aliphatic, alicyclic or aromatic organic amines such as isopropylamine, methylamine, trimethylamine, picoline, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2- diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Exemplary organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. The selection of the appropriate salt may be useful, for example, so that an ester functionality, if any, elsewhere in a compound is not hydrolyzed. The selection criteria for the appropriate salt will be known to one skilled in the art.

[0130] Solvates of compounds of the application include, for example, those made with solvents that are pharmaceutically acceptable. Examples of such solvents include water (resulting solvate is called a hydrate) and ethanol and the like. Suitable solvents are physiologically tolerable at the dosage administered.

[0131] In some embodiments of the present application, the compounds described herein have at least one asymmetric center. Where compounds possess more than one asymmetric center, they may exist as diastereomers. It is to be understood that all such isomers and mixtures thereof in any proportion are encompassed within the scope of the present application. It is to be further that while the stereochemistry of the 25 8713541compounds may be as shown in any given compound listed herein, such compounds may also contain certain amounts (for example, less than 20%, suitably less than 10%, more suitably less than 5%) of compounds of the present application having an alternate stereochemistry. It is intended that any optical isomers, as separated, pure or partially purified optical isomers or racemic mixtures thereof are included within the scope of the present application.

[0132] The compounds of the present application may also exist in different tautomeric forms and it is intended that any tautomeric forms which the compounds form, as well as mixtures thereof, are included within the scope of the present application.

[0133] The compounds of the present application may further exist in varying polymorphic forms and it is contemplated that any polymorphs, or mixtures thereof, which form are included within the scope of the present application.

[0134] The compounds of the present application may further be radiolabeled and accordingly all radiolabeled versions of the compounds of the application are included within the scope of the present application. The compounds of the application also include those in which one or more radioactive atoms are incorporated within their structure. III. Compositions of the Application

[0135] The compounds of the present application are suitably formulated in a conventional manner into compositions using one or more carriers. Accordingly, the present application also includes a composition comprising one or more compounds of the application and a carrier. The compounds of the application are suitably formulated into pharmaceutical compositions for administration to subjects in a biologically compatible form suitable for administration in vivo. Accordingly, the present application further includes a pharmaceutical composition comprising one or more compounds of the application and a pharmaceutically acceptable carrier. In embodiments of the application the pharmaceutical compositions are used in the treatment of any of the diseases, disorders or conditions described herein.

[0136] The compounds of the application are administered to a subject in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. For example, a compound of the application is administered by oral, inhalation, parenteral, buccal, sublingual, nasal, rectal, vaginal, patch, pump, minipump, topical or transdermal administration and the pharmaceutical compositions formulated accordingly. In some embodiments, is by means of a pump for periodic or 26 8713541continuous delivery. Conventional procedures and ingredients for the selection and preparation of suitable compositions are described, for example, in Remington’s Pharmaceutical Sciences (2000 - 20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999.

[0137] Parenteral administration includes systemic delivery routes other than the gastrointestinal (GI) tract, and includes, for example intravenous, intra-arterial, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary (for example, by use of an aerosol), intrathecal, rectal and topical (including the use of a patch or other transdermal delivery device) modes of administration. Parenteral administration may be by continuous infusion over a selected period of time.

[0138] In some embodiments, a compound of the application is orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it is enclosed in hard or soft shell gelatin capsules, or it is compressed into tablets, or it is incorporated directly with the food of the diet. In some embodiments, the compound is incorporated with excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, caplets, pellets, granules, lozenges, chewing gum, powders, syrups, elixirs, wafers, aqueous solutions and suspensions, and the like. In the case of tablets, carriers that are used include lactose, corn starch, sodium citrate and salts of phosphoric acid. Pharmaceutically acceptable excipients include binding agents (e.g., pregelatinized maize 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 starch glycolate); or wetting agents (e.g., sodium lauryl sulphate). In embodiments, the tablets are coated by methods well known in the art. In the case of tablets, capsules, caplets, pellets or granules for oral administration, pH sensitive enteric coatings, such as Eudragits™ designed to control the release of active ingredients are optionally used. Oral dosage forms also include modified release, for example immediate release and timed-release, formulations. Examples of modified-release formulations include, for example, sustained-release (SR), extended- release (ER, XR, or XL), time-release or timed-release, controlled-release (CR), or continuous-release (CR or Contin), employed, for example, in the form of a coated tablet, an osmotic delivery device, a coated capsule, a microencapsulated microsphere, an agglomerated particle, e.g., as of molecular sieving type particles, or, a fine hollow permeable fiber bundle, or chopped hollow permeable fibers, agglomerated or held in a fibrous packet. Timed-release formulated, for example as liposomes or 27 8713541those wherein the active compound is protected with differentially degradable coatings, such as by microencapsulation, multiple coatings, etc. Liposome delivery systems include, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. In some embodiments, liposomes are formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. For oral administration in a capsule form, useful carriers or diluents include lactose and dried corn starch.

[0139] In some embodiments, liquid preparations for oral administration take the form of, for example, solutions, syrups or suspensions, or they are suitably presented as a dry product for constitution with water or other suitable vehicle before use. When aqueous suspensions and / or emulsions are administered orally, the compound of the application is suitably suspended or dissolved in an oily phase that is combined with emulsifying and / or suspending agents. If desired, certain sweetening and / or flavoring and / or coloring agents are added. Such liquid preparations for oral administration are prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid). Useful diluents include lactose and high molecular weight polyethylene glycols.

[0140] It is also possible to freeze-dry the compounds of the application and use the lyophilizates obtained, for example, for the preparation of products for injection.

[0141] In some embodiments, a compound of the application is administered parenterally. For example, solutions of a compound of the application are prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. In some embodiments, dispersions are prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. A person skilled in the art would know how to prepare suitable formulations. For parenteral administration, sterile solutions of the compounds of the application are usually prepared, and the pH’s of the solutions are suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic. For ocular administration, ointments or droppable liquids are delivered, for example, by ocular delivery systems known to the art such as applicators or eye droppers. In some embodiment, such compositions include mucomimetics such as hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose or polyvinyl alcohol, preservatives such 28 8713541as sorbic acid, EDTA or benzyl chromium chloride, and the usual quantities of diluents or carriers. For pulmonary administration, diluents or carriers will be selected to be appropriate to allow the formation of an aerosol.

[0142] In some embodiments, a compound of the application is formulated for parenteral administration by injection, including using conventional catheterization techniques or infusion. Formulations for injection are, for example, presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. In some embodiments, the compositions take such forms as sterile suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulating agents such as suspending, stabilizing and / or dispersing agents. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. Alternatively, the compounds of the application are suitably in a sterile powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0143] In some embodiments, compositions for nasal administration are conveniently formulated as aerosols, drops, gels and powders. For intranasal administration or administration by inhalation, the compounds of the application are conveniently delivered in the form of a solution, dry powder formulation or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which, for example, take the form of a cartridge or refill for use with an atomising device. Alternatively, the sealed container is a unitary dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant which is, for example, a compressed gas such as compressed air or an organic propellant such as fluorochlorohydrocarbon. Suitable propellants include but are not limited to dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, heptafluoroalkanes, carbon dioxide or another suitable gas. In the case of a pressurized aerosol, the dosage unit is suitably determined by providing a valve to deliver a metered amount. In some embodiments, the pressurized container or nebulizer contains a solution or suspension of the active compound. Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator example, formulated containing a powder 29 8713541mix of a compound of the application and a suitable powder base such as lactose or starch. The aerosol dosage forms can also take the form of a pump-atomizer.

[0144] Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein a compound of the application is formulated with a carrier such as sugar, acacia, tragacanth, or gelatin and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0145] Suppository forms of the compounds of the application are useful for vaginal, urethral and rectal administrations. Such suppositories will generally be constructed of a mixture of substances that is solid at room temperature but melts at body temperature. The substances commonly used to create such vehicles include but are not limited to theobroma oil (also known as cocoa butter), glycerinated gelatin, other glycerides, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol. See, for example: Remington's Pharmaceutical Sciences, 16th Ed., Mack Publishing, Easton, PA, 1980, pp. 1530-1533 for further discussion of suppository dosage forms.

[0146] In some embodiments a compound of the application is coupled with soluble polymers as targetable drug carriers. Such polymers include, for example, polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-ethylaspartamide-phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, in some embodiments, a compound of the application is coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.

[0147] In some embodiments, compounds of the application may be coupled with viral, non-viral or other vectors. Viral vectors may include retrovirus, lentivirus, adenovirus, herpesvirus, poxvirus, alphavirus, vaccinia virus or adeno-associated viruses. Non-viral vectors may include nanoparticles, cationic lipids, cationic polymers, metallic nanoparticles, nanorods, liposomes, micelles, microbubbles, cell-penetrating peptides, or lipospheres. Nanoparticles may include silica, lipid, carbohydrate, or other pharmaceutically acceptable polymers. 30 8713541

[0148] A compound of the application including pharmaceutically acceptable salts and / or solvates thereof is suitably used on their own but will generally be administered in the form of a pharmaceutical composition in which the one or more compounds of the application (the active ingredient) is in association with a pharmaceutically acceptable carrier. Depending on the mode of administration, the pharmaceutical composition will comprise from about 0.05 wt% to about 99 wt% or about 0.10 wt% to about 70 wt%, of the active ingredient, and from about 1 wt% to about 99.95 wt% or about 30 wt% to about 99.90 wt% of a pharmaceutically acceptable carrier, all percentages by weight being based on the total composition. IV. Methods and Uses of the Application

[0149] The compounds of the application have been shown to inhibit or block general control nonderepressible 2 (GCN2) kinase, to attenuate the transcriptional function of ATF4 target gene expression. Therefore, the compounds of the application are useful for inhibiting GCN2.

[0150] Accordingly, the present application includes a method of inhibiting general control nonderepressible 2 (GCN2) in a cell, either in a biological sample or in a patient, comprising administering an effective amount of one or more compounds of the application to the cell.

[0151] The present application also includes a use of one or more compounds of the application for inhibiting GCN2 in a cell as well as a use of one or more compounds of the application for the preparation of a medicament for inhibiting GCN2 in a cell. The application further includes one or more compounds of the application for use in inhibiting GCN2 in a cell.

[0152] As the compounds of the application have been shown to be capable of inhibiting GCN2 protein activity, the compounds of the application are useful for treating diseases, disorders or conditions by inhibiting GCN2. Therefore, the compounds of the present application are useful as medicaments. Accordingly, the present application includes a compound of the application for use as a medicament.

[0153] Accordingly, the present application also includes a method of treating a disease, disorder or condition that is treatable by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof. 31 8713541

[0154] The present application also includes a use of one or more compounds of the application for treatment of a disease, disorder or condition that is treatable by inhibiting GCN2, as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of a disease, disorder or condition that is treatable by inhibiting GCN2. The application further includes one or more compounds of the application for use in treating a disease, disorder or condition that is treatable by inhibiting GCN2.

[0155] “GCN2” is a protein kinase that belongs to the family of eukaryotic initiation factor 2 α (eIF2α) kinases. In some embodiments, this serine / threonine-protein kinase is an enzyme that in humans is encoded by the GCN2 or EIF2AK4 (Gene ID: 851877) comprising the amino acid sequence disclosed in Mol. Cell. Biol.1995,15 (8): 4497–506.

[0156] In some embodiments, the disease, disorder or condition that is treatable by inhibiting GCN2 is a neoplastic disorder. Accordingly, the present application also includes a method of treating a neoplastic disorder comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof. The present application also includes a use of one or more compounds of the application for treatment of a neoplastic disorder as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of a neoplastic disorder. The application further includes one or more compounds of the application for use in treating a neoplastic disorder. In some embodiments, the treatment is in an amount effective to ameliorate at least one symptom of the neoplastic disorder, for example, reduced cell proliferation or reduced tumor mass, among others, in a subject in need of such treatment.

[0157] Neoplasms can be benign (such as uterine fibroids and melanocytic nevi), potentially malignant (such as carcinoma in situ) or malignant (i.e. cancer). Exemplary neoplastic disorders include the so-called solid tumours and liquid tumours, including but not limited to carcinoma, sarcoma, metastatic disorders (e.g., tumors arising from the prostate), hematopoietic neoplastic disorders, (e.g., leukemias, lymphomas, myeloma and other malignant plasma cell disorders), metastatic tumors and other cancers. Prevalent cancers include breast, prostate, colon, lung, liver, brain, ovarian and pancreatic cancers.

[0158] Compounds of the application have been demonstrated to inhibit the growth of cancer cells. In some embodiments, the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer.

[0159] Accordingly, the present application also includes a method of treating cancer comprising administering a effective amount of one or more 32 8713541compounds of the application to a subject in need thereof. The present application also includes a use of one or more compounds of the application for treatment of cancer as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of cancer. The application further includes one or more compounds of the application for use in treating cancer. In an embodiment, the compound is administered for the prevention of cancer in a subject such as a mammal having a predisposition for cancer.

[0160] In some embodiments, the cancer is selected from, but not limited to: Acute Lymphoblastic Leukemia, Adult; Acute Lymphoblastic Leukemia, Childhood; Acute Myeloid Leukemia, Adult; Adrenocortical Carcinoma; Adrenocortical Carcinoma, Childhood; AIDS- Related Lymphoma; AIDS-Related Malignancies; Anal Cancer; Astrocytoma, Childhood Cerebellar; Astrocytoma, Childhood Cerebral; Bile Duct Cancer, Extrahepatic; Bladder Cancer; Bladder Cancer, Childhood; Bone Cancer, Osteosarcoma / Malignant Fibrous Histiocytoma; Brain Stem Glioma, Childhood; Brain Tumor, Adult; Brain Tumor, Brain Stem Glioma, Childhood; Brain Tumor, Cerebellar Astrocytoma, Childhood; Brain Tumor, Cerebral Astrocytoma / Malignant Glioma, Childhood; Brain Tumor, Ependymoma, Childhood; Brain Tumor, Medulloblastoma, Childhood; Brain Tumor, Supratentorial Primitive Neuroectodermal Tumors, Childhood; Brain Tumor, Visual Pathway and Hypothalamic Glioma, Childhood; Brain Tumor, Childhood (Other); Breast Cancer; Breast Cancer and Pregnancy; Breast Cancer, Childhood; Breast Cancer, Male; Bronchial Adenomas / Carcinoids, Childhood; Carcinoid Tumor, Childhood; Carcinoid Tumor, Gastrointestinal; Carcinoma, Adrenocortical; Carcinoma, Islet Cell; Carcinoma of Unknown Primary; Central Nervous System Lymphoma, Primary; Cerebellar Astrocytoma, Childhood; Cerebral Astrocytoma / Malignant Glioma, Childhood; Cervical Cancer; Childhood Cancers; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia; Chronic Myeloproliferative Disorders; Clear Cell Sarcoma of Tendon Sheaths; Colon Cancer; Colorectal Cancer, Childhood; Cutaneous T-CeIl Lymphoma; Endometrial Cancer; Ependymoma, Childhood; Epithelial Cancer, Ovarian; Esophageal Cancer; Esophageal Cancer, Childhood; Ewing's Family of Tumors; Extracranial Germ Cell Tumor, Childhood; Extragonadal Germ Cell Tumor; Extrahepatic Bile Duct Cancer; Eye Cancer, Intraocular Melanoma; Eye Cancer, Retinoblastoma; Gallbladder Cancer; Gastric (Stomach) Cancer; Gastric (Stomach) Cancer, Childhood; Gastrointestinal Carcinoid Tumor; Germ Cell Tumor, Extracranial, Childhood; Germ Cell Tumor, Extragonadal; Germ Cell Tumor, Ovarian; Gestational Trophoblastic Tumor; Glioma, Childhood Brain Stem; Glioma, Childhood Visual Pathway and Hypothalamic; Hairy Cell Head and Neck Cancer; Hepatocellular 33 8713541(Liver) Cancer, Adult (Primary); Hepatocellular (Liver) Cancer, Childhood (Primary); Hodgkin's Lymphoma, Adult; Hodgkin's Lymphoma, Childhood; Hodgkin's Lymphoma During Pregnancy; Hypopharyngeal Cancer; Hypothalamic and Visual Pathway Glioma, Childhood; Intraocular Melanoma; Islet Cell Carcinoma (Endocrine Pancreas); Kaposi's Sarcoma; Kidney Cancer; Laryngeal Cancer; Laryngeal Cancer, Childhood; Leukemia, Acute Lymphoblastic, Adult; Leukemia, Acute Lymphoblastic, Childhood; Leukemia, Acute Myeloid, Adult; Leukemia, Acute Myeloid, Childhood; Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Cell; Lip and Oral Cavity Cancer; Liver Cancer, Adult (Primary); Liver Cancer, Childhood (Primary); Lung Cancer, Non-Small Cell; Lung Cancer, Small Cell; Lymphoblastic Leukemia, Adult Acute; Lymphoblastic Leukemia, Childhood Acute; Lymphocytic Leukemia, Chronic; Lymphoma, AIDS-Related; Lymphoma, Central Nervous System (Primary); Lymphoma, Cutaneous T-CeIl; Lymphoma, Hodgkin's, Adult; Lymphoma, Hodgkin's, Childhood; Lymphoma, Hodgkin's During Pregnancy; Lymphoma, Non-Hodgkin's, Adult; Lymphoma, Non-Hodgkin's, Childhood; Lymphoma, Non-Hodgkin's During Pregnancy; Lymphoma, Primary Central Nervous System; Macroglobulinemia, Waldenstrom's; Male Breast Cancer; Malignant Mesothelioma, Adult; Malignant Mesothelioma, Childhood; Malignant Thymoma; Medulloblastoma, Childhood; Melanoma; Melanoma, Intraocular; Merkel Cell Carcinoma; Mesothelioma, Malignant; Metastatic Squamous Neck Cancer with Occult Primary; Multiple Endocrine Neoplasia Syndrome, Childhood; Multiple Myeloma / Plasma Cell Neoplasm; Mycosis Fungoides; Myelodysplastic Syndromes; Myelogenous Leukemia, Chronic; Myeloid Leukemia, Childhood Acute; Myeloma, Multiple; Myeloproliferative Disorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer; Nasopharyngeal Cancer; Nasopharyngeal Cancer, Childhood; Neuroblastoma; Non-Hodgkin's Lymphoma, Adult; Non-Hodgkin's Lymphoma, Childhood; Non- Hodgkin's Lymphoma During Pregnancy; Non-Small Cell Lung Cancer; Oral Cancer, Childhood; Oral Cavity and Lip Cancer; Oropharyngeal Cancer; Osteosarcoma / Malignant Fibrous Histiocytoma of Bone; Ovarian Cancer, Childhood; Ovarian Epithelial Cancer; Ovarian Germ Cell Tumor; Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; Pancreatic Cancer, Childhood; Pancreatic Cancer, Islet Cell; Paranasal Sinus and Nasal Cavity Cancer; Parathyroid Cancer; Penile Cancer; Pheochromocytoma; Pineal and Supratentorial Primitive Neuroectodermal Tumors, Childhood; Pituitary Tumor; Plasma Cell Neoplasm / Multiple Myeloma; Pleuropulmonary Blastoma; Pregnancy and Breast Cancer; Pregnancy and Hodgkin's Lymphoma; Pregnancy and Non-Hodgkin's Lymphoma; Primary Central Nervous System Lymphoma; Primary Liver Cancer, Adult; Primary Liver Cancer, Childhood; Prostate Cancer; Rectal 34 8713541Cancer; Renal Cell (Kidney) Cancer; Renal Cell Cancer, Childhood; Renal Pelvis and Ureter, Transitional Cell Cancer; Retinoblastoma; Rhabdomyosarcoma, Childhood; Salivary Gland Cancer; Salivary Gland Cancer, Childhood; Sarcoma, Ewing's Family of Tumors; Sarcoma, Kaposi's; Sarcoma (Osteosarcoma) / Malignant Fibrous Histiocytoma of Bone; Sarcoma, Rhabdomyosarcoma, Childhood; Sarcoma, Soft Tissue, Adult; Sarcoma, Soft Tissue, Childhood; Sezary Syndrome; Skin Cancer; Skin Cancer, Childhood; Skin Cancer (Melanoma); Skin Carcinoma, Merkel Cell; Small Cell Lung Cancer; Small Intestine Cancer; Soft Tissue Sarcoma, Adult; Soft Tissue Sarcoma, Childhood; Squamous Neck Cancer with Occult Primary, Metastatic; Stomach (Gastric) Cancer; Stomach (Gastric) Cancer, Childhood; Supratentorial Primitive Neuroectodermal Tumors, Childhood; T-CeIl Lymphoma, Cutaneous; Testicular Cancer; Thymoma, Childhood; Thymoma, Malignant; Thyroid Cancer; Thyroid Cancer, Childhood; Transitional Cell Cancer of the Renal Pelvis and Ureter; Trophoblastic Tumor, Gestational; Unknown Primary Site, Cancer of, Childhood; Unusual Cancers of Childhood; Ureter and Renal Pelvis, Transitional Cell Cancer; Urethral Cancer; Uterine Sarcoma; Vaginal Cancer; Visual Pathway and Hypothalamic Glioma, Childhood; Vulvar Cancer; Waldenstrom's Macro globulinemia; and Wilms' Tumor. Metastases of the aforementioned cancers can also be treated in accordance with the methods described herein.

[0161] In some embodiments, the cancer is any cancer in which the cells show increased expression of the gene(s) encoding GCN2 or activation of GCN2 under stress conditions. By “increased expression” it is meant any increase in expression of the gene(s) encoding GCN2 in the cell compared to expression of the gene(s) encoding GCN2 in a corresponding normal or healthy cell.

[0162] 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. 35 8713541

[0163] In some embodiments, the disease, disorder or condition that is treatable by inhibiting GCN2, is a disease, disorder or condition associated with an uncontrolled and / or abnormal cellular activity affected directly or indirectly by inhibiting GCN2. In another embodiment, the uncontrolled and / or abnormal cellular activity that is affected directly or indirectly by inhibiting GCN2 is proliferative activity in a cell.

[0164] Accordingly, the application also includes a method of inhibiting proliferative activity in a cell, comprising administering an effective amount of one or more compounds of the application to the cell. The present application also includes a use of one or more compounds of the application for inhibition of proliferative activity in a cell as well as a use of one or more compounds of the application for the preparation of a medicament for inhibition of proliferative activity in a cell. The application further includes one or more compounds of the application for use in inhibiting proliferative activity in a cell.

[0165] The present application also includes a method of inhibiting uncontrolled and / or abnormal cellular activities affected directly or indirectly by inhibiting GCN2 in a cell, either in a biological sample or in a subject, comprising administering an effective amount of one or more compounds of the application to the cell. The application also includes a use of one or more compounds of the application for inhibition of uncontrolled and / or abnormal cellular activities affected directly or indirectly by inhibiting GCN2 in a cell as well as a use of one or more compounds of the application for the preparation of a medicament for inhibition of uncontrolled and / or abnormal cellular activities affected directly or indirectly by inhibiting GCN2 in a cell. The application further includes one or more compounds of the application for use in inhibiting uncontrolled and / or abnormal cellular activities affected directly or indirectly by inhibiting GCN2 in a cell.

[0166] In some embodiments, the disease, disorder or condition that is treatable by inhibiting GCN2 is a peripheral neuropathy. Accordingly, the present application also includes a method of treating a peripheral neuropathy comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof. The present application also includes a use of one or more compounds of the application for treatment of a peripheral neuropathy as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of a peripheral neuropathy. The application further includes one or more compounds of the application for use in treating a peripheral neuropathy. 36 8713541

[0167] In some embodiments, the peripheral neuropathy is Charcot-Marie-Tooth (CMT) peripheral neuropathy. Heterozygous mutations in six genes encoding cytoplasmic aminoacyl-tRNA synthetases (AARSs) cause axonal and intermediate forms of CMT peripheral neuropathy. Heterozygous mutations in six genes encoding cytoplasmic aminoacyl-tRNA synthetases (AARSs) cause axonal and intermediate forms of Charcot- Marie-Tooth (CMT) peripheral neuropathy. AARSs are ubiquitously expressed enzymes that covalently attach amino acids to their cognate tRNAs (tRNA aminoacylation). Aminoacylated tRNAs are used by the ribosome for mRNA translation. In Charcot-Marie- Tooth (CMT) peripheral neuropathy, mutant tRNA synthetases activate the integrated stress response (ISR) through the sensor kinase GCN2. The chronic activation of the ISR contributed to the pathophysiology, and genetic deletion or pharmacological inhibition of GCN2 alleviated the peripheral neuropathy. Therefore, in some embodiments, the disease, disorder or condition that is treatable by inhibiting GCN2, is a Charcot-Marie-Tooth (CMT) peripheral neuropathy.

[0168] Accordingly, the present application also includes a method of treating Charcot-Marie-Tooth (CMT) peripheral neuropathy comprising administering to a subject in need thereof, a therapeutically effective amount of one or more compounds of the application. The present application also includes a use of one or more compounds of the application for treatment of Charcot-Marie-Tooth (CMT) peripheral neuropathy as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of Charcot-Marie-Tooth (CMT) peripheral neuropathy. The application further includes one or more compounds of the application for use in treating Charcot-Marie-Tooth (CMT) peripheral neuropathy.

[0169] The present application also includes a method of treating a disease, disorder or condition that is treatable by inhibiting GCN2 comprising administering a therapeutically effective amount of one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibiting GCN2 to a subject in need thereof. The present application also includes a use of one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibiting GCN2, as well as a use of one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibiting GCN2 for the preparation of a medicament for treatment of a disease, disorder or condition treatable inhibiting GCN2. The application further 37 8713541includes one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibiting GCN2 for use in treating a disease, disorder or condition treatable by inhibiting GCN2.

[0170] In an embodiment, the disease, disorder or condition treatable by inhibiting GCN2 is cancer and / or peripheral neuropathy.

[0171] In some embodiments, GCN2 is inhibited in the uses and methods of the application.

[0172] In an embodiment, the "subject in need thereof” is a subject having the disease, disorder or condition to be treated.

[0173] In an embodiment, the subject is a mammal. In another embodiment, the subject is human.

[0174] In some embodiments the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer and the one or more compounds of the application are administered or used in combination with one or more additional cancer treatments. In another embodiment, the one or more additional cancer treatments is selected from one or more radiotherapy, chemotherapy, targeted therapies such as antibody therapies (including anti-PD1 and / or anti-PD-L1 antibodies) and small molecule therapies such as tyrosine-kinase inhibitors therapies, glutaminase inhibitors (e.g., glutaminase-1 (GLS1) inhibitors), and asparagine synthetase (ASNS) inhibitors, immunotherapy, hormonal therapy and anti-angiogenic therapies.

[0175] 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 is treatable by inhibiting GCN2 is cancer, and the one or more compounds of the 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 is treatable by inhibiting GCN2 is cancer, and the one or more compounds of the application are administered or used in combination with L-asparaginase (L-ASNase).

[0176] In some embodiments, the small molecule therapy is a glutaminase (e.g., glutaminase-1, (GLS1)) inhibitor or an asparagine synthetase (ASNS) inhibitor. Therefore, in some embodiments the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer and the one or more compounds of the application are administered or 38 8713541used in combination one or more glutaminase inhibitors (e.g., GLS1 inhibitors), and / or or asparagine synthetase (ASNS) inhibitors.

[0177] In some embodiments the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer and the one or more compounds of the application are administered or used in combination one or more glutaminase inhibitors (e.g., GLS1 inhibitors), and / or asparagine synthetase (ASNS) inhibitors and / or L-asparaginase (L- ASNase).

[0178] Asparagine deprivation by L-asparaginase (L-ASNase) is an effective therapeutic strategy in cancer, with resistance occurring due to upregulation of asparagine synthetase (ASNS), the only human enzyme synthetizing asparagine (Annu. Rev. Biochem.2006, 75 (1), 629–654). L-Asparaginase efficacy in solid tumors is limited by dose-related toxicities (OncoTargets and Therapy 2017, pp 1413–1422). Large-scale loss of function genetic in vitro screens identified ASNS as a cancer dependency in several solid malignancies (Cell 2017, 170 (3), 564–576.e16. Cell 2017, 170 (3), 577–592.e10). Genome-wide CRISPR screens have revealed that cancer cell resistance mechanisms are elicited by a GCN2-ATF4 axis aimed at restoring amino acid levels to promote survival. Hence, pharmacological inhibition of GCN2 synergizes with L-asparaginase-mediated asparagine deprivation in ASNS deficient cells suggesting novel potential therapeutic combinations in cancer treatment.

[0179] Therefore, in some embodiments, the present application also includes a method of improving the efficacy of one or more cancer treatments for treating cancer comprising administering an effective amount of one or more compounds of the application in combination with an effective amount of the one or more cancer treatments to a subject in need thereof.

[0180] The present application also includes a use of one or more compounds of the application in combination with one or more cancer treatments for improving the efficacy of the one or more cancer treatments for treating cancer, as well as a use of one or more compounds of the application in combination with one or more cancer treatments for improving the efficacy of the one or more cancer treatments for treating cancer. The application further includes one or more compounds of the application in combination with one or more cancer treating for use in improving the efficacy of the one or more cancer treatments for treating cancer. 39 8713541

[0181] In some embodiments, the one or more cancer treatments is selected from one or more radiotherapy, chemotherapy, targeted therapies such as antibody therapies (including anti-PD1 and / or anti-PD-L1 antibodies) and small molecule therapies such as tyrosine-kinase inhibitors therapies, glutaminase inhibitors (e.g.,GLS1 inhibitors), and / or asparagine synthetase (ASNS) inhibitors, immunotherapy, hormonal therapy and anti- angiogenic therapies.

[0182] In some embodiments, the chemotherapy is a chemotherapeutic agent. In some embodiments, chemotherapeutic agent is cisplatin. Therefore, in some embodiments, the one or more compounds of the application are administered or used in combination with cisplatin for improving the efficacy of cisplatin for treating cancer.

[0183] In some embodiments, the chemotherapeutic agent is L-asparaginase (L- ASNase). Therefore, in some embodiments, the one or more compounds of the application are administered or used in combination with L-asparaginase (L-ASNase) for improving the efficacy of L-ASNase for treating cancer.

[0184] In some embodiments, the small molecule therapy is a glutaminase inhibitor (e.g., GLS1 inhibitor) or an asparagine synthetase (ASNS) inhibitor. Therefore, in some embodiments, the one or more compounds of the application are administered or used in combination with one or more glutaminase inhibitors (e.g.,GLS1 inhibitors) and / or one or more asparagine synthetase (ASNS) inhibitors for improving the efficacy the one or more glutaminase inhibitors (e.g.,GLS1 inhibitors) or the one or more ASNS inhibitors for treating cancer.

[0185] In some embodiments, the cancer is associated with low asparagine synthetase (ASNS) expression. In some embodiment, the cancer is associated with low asparagine synthetase (ASNS) expression, and the chemotherapeutic agent is L- asparaginase (L-ASNase). Accordingly, in some embodiments, the one or more compounds of the application are administered or used in combination with L-asparaginase (L-ASNase) for improving the efficacy of L-ASNase for treating a cancer is associated with low asparagine synthetase (ASNS) expression.

[0186] In some embodiments, the cancer is associated with asparagine synthetase (ASNS) overexpression or dysregulation. In some embodiments, the cancer is associated with asparagine synthetase (ASNS) overexpression or dysregulation and the chemotherapeutic agents are one or more asparagine synthetase (ASNS) inhibitors and / or L-asparaginase Therefore, in some the one or more compounds of the 40 8713541application are administered or used in combination with one or more asparagine synthetase (ASNS) inhibitors and / or with L-asparaginase for treating a cancer associated with asparagine synthetase (ASNS) overexpression or dysregulation. In some embodiments, the chemotherapeutic agents are one or more asparagine synthetase (ASNS) inhibitors and L-asparaginase

[0187] In some embodiments, the cancer is associated with low asparagine synthetase (ASNS) expression and low glutaminase (e.g. GLS1) expression. In some embodiments, the cancer is associated with low asparagine synthetase (ASNS) expression and low glutaminase (e.g., GLS1) expression and the chemotherapeutic agents are L- asparaginase (L-ASNase) and / or one or more glutaminase inhibitors. Accordingly, in some embodiments, the one or more compounds of the application are administered or used in combination with L-asparaginase (L-ASNase) and / or one or more glutaminase inhibitors for improving the efficacy of L-ASNase and / or the one or more glutaminase inhibitors for treating a cancer associated with low asparagine synthetase (ASNS) expression and low glutaminase (e.g. GLS1) expression. In some embodiments, the glutaminase inhibitor is a GLS1 inhibitor. In some embodiments, the chemotherapeutic agents are L-asparaginase (L-ASNase) and one or more glutaminase inhibitors.

[0188] In some embodiments, the cancer is associated with asparagine synthetase (ASNS) overexpression or dysregulation and glutaminase (e.g. GLS1) overexpression or dysregulation. In some embodiments, the cancer is associated with asparagine synthetase (ASNS) overexpression or dysregulation and glutaminase (e.g. GLS1) overexpression or dysregulation and the chemotherapeutic agents are L-asparaginase (L-ASNase), one or more glutaminase inhibitors and / or one or more asparagine synthetase (ASNS) inhibitors. Therefore, in some embodiments, the one or more compounds of the application are administered or used in combination with L-asparaginase (L-ASNase) and / or one or more glutaminase inhibitors and / or one or more asparagine synthetase (ASNS) inhibitors for improving the efficacy of L-ASNase and / or the one or more glutaminase inhibitors and / or the one or more asparagine synthetase (ASNS) inhibitors for treating a cancer associated with asparagine synthetase (ASNS) overexpression or dysregulation and glutaminase (e.g. GLS1) overexpression or dysregulation. In some embodiments, the glutaminase inhibitors is a GLS1 inhibitor. In some embodiments, the chemotherapeutic agents are L- asparaginase (L-ASNase), one or more glutaminase inhibitors and one or more asparagine synthetase (ASNS) inhibitors. 41 8713541

[0189] Compounds of the application are either used alone or in combination with other known agents useful for treating diseases, disorders or conditions treatable by inhibiting GCN2. When used in combination with other agents useful in treating diseases, disorders or conditions that are treatable by inhibiting GCN2, it is an embodiment that the compounds of the application are administered contemporaneously with those agents. As used herein, “contemporaneous administration” of two substances to a subject means providing each of the two substances so that they are both biologically active in the individual at the same time. The exact details of the administration will depend on the pharmacokinetics of the two substances in the presence of each other and can include administering the two substances within a few hours of each other, or even administering one substance within 24 hours of administration of the other, if the pharmacokinetics are suitable. Design of suitable dosing regimens is routine for one skilled in the art. In particular embodiments, two substances will be administered substantially simultaneously, i.e., within minutes of each other, or in a single composition that contains both substances. It is a further embodiment of the present application that a combination of agents is administered to a subject in a non-contemporaneous fashion. In some embodiments, compounds of the present application are administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present application provides a single unit dosage form comprising one or more compounds of the application (e.g., a compound of Formula I), an additional therapeutic agent, and a pharmaceutically acceptable carrier.

[0190] Treatment methods comprise administering to a subject a therapeutically effective amount of one or more of the compounds of the application and optionally consist of a single administration, or alternatively comprise a series of administrations, and optionally comprise concurrent administration or use of one or more other therapeutic agents. For example, in some embodiments, the compounds of the application are administered at least once a week. In some embodiments, the compounds are administered to the subject from about one time per two or three weeks, or about one time per week to about once daily for a given treatment. In another embodiment, the compounds are 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 condition, the age of the subject, the concentration and / or the activity of the compounds of the application, and / or a combination thereof. It will also be appreciated that the effective dosage of the compound used for the treatment may increase or decrease over the course of a particular 42 8713541treatment regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. For example, the compounds are administered to the subject in an amount and for duration sufficient to treat the subject. In some embodiments treatment comprise prophylactic treatment. For example, a subject with early cancer can be treated to prevent progression, or alternatively a subject in remission can be treated with a compound or composition of the application to prevent recurrence.

[0191] The dosage of compounds of the application varies depending on many factors such as the pharmacodynamic properties of the compound, the mode of administration, the age, health and weight of the recipient, the nature and extent of the symptoms, the frequency of the treatment and the type of concurrent treatment, if any, and the clearance rate of the compound in the subject to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. Compounds of the application may be administered initially in a suitable dosage that may be adjusted as required, depending on the clinical response. Dosages will generally be selected to maintain a serum level of compounds of the application from 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, oral dosages of one or more compounds of the application will range between about 0.05 mg per day to about 1000 mg per day for an adult, suitably about 0.1 mg per day to about 500 mg per day, more suitably about 1 mg per day to about 200 mg per day. For parenteral administration, a representative amount is from 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 will be administered. For oral administration, a representative amount is from 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 administration in suppository form, a representative amount is from about 0.1 mg / kg to about 10 mg / kg or about 0.1 mg / kg to about 1 mg / kg. Compounds of the application may be administered in a single daily, weekly or monthly dose or the total daily dose may be divided into two, three or four daily doses.

[0192] In an embodiment, effective amounts vary according to factors such as the disease state, age, sex and / or weight of the subject. In a further embodiment, the amount of a given compound or compounds that will correspond to an effective amount will vary depending upon factors, such as the given drug(s) or compound(s), the pharmaceutical formulation, the route of administration, type of condition, disease or disorder, the 43 8713541identity of the subject being treated, and the like, but can nevertheless be routinely determined by one skilled in the art.

[0193] To be clear, in the above, the term “a compound” also includes embodiments wherein one or more compounds are referenced. Likewise, the term “compounds of the application” also includes embodiments wherein only one compound is referenced. V. Methods of Preparation of Compounds of the Application

[0194] Compounds of the present application can be prepared by various synthetic processes. The choice of particular structural features and / or substituents may influence the selection of one process over another. The selection of a particular process to prepare a given compound of Formula I is within the purview of the person of skill in the art. Some starting materials for preparing compounds of the present application are available from commercial chemical sources. Other starting materials, for example as described below, are readily prepared from available precursors using straightforward transformations that are well known in the art.

[0195] The compounds of Formula I generally can be prepared according to the processes illustrated in the Schemes below. In the structural formulae shown below the variables are as defined in Formula I unless otherwise stated. A person skilled in the art would appreciate that many of the reactions depicted in the Schemes below would be sensitive to oxygen and water and would know to perform the reaction under an anhydrous, inert atmosphere if needed. Reaction temperatures and times are presented for illustrative purposes only and may be varied to optimize yield as would be understood by a person skilled in the art.

[0196] Accordingly, in some embodiments, the compounds of Formula I are prepared as shown in Scheme 1. 44 8713541Scheme 1

[0197] In some embodiments shown in Scheme 1, compounds of Formula I are prepared by oxidation of compounds of Formula A with suitable oxidizing agents such as meta-chloroperoxybenzoic acid (m-CPBA) to the corresponding sulfoxide followed by substitution with various suitable amines compounds of Formula B to provide the bromo- intermediates of Formula C. Subsequent coupling of compounds of Formula C with suitable boronates of Formula D (wherein Raand Rbare independently C1-6alkyl, or are joined to form, together with the B and O atoms therebetween, a 4 to 6 membered saturated or unsaturated ring optionally substituted with one or two C1-3alkyls) under suitable coupling conditions such as Suzuki-Miyaura coupling conditions afford compounds of Formula I.Scheme 2

[0198] In some embodiments, compounds of Formula I are prepared as shown in Scheme 2. Therefore the intermediate compounds of Formula A are coupled with boronate compounds of Formula D under suitable coupling conditions such as under Suzuki-Miyaura coupling conditions to provide intermediate compounds of Formula E. Oxidation of compounds of Formula E to the corresponding sulfoxide with suitable oxidizing agents such 45 8713541as meta-chloroperoxybenzoic acid (m-CPBA) followed by substitution with various suitable amines of Formula B provides the compounds of Formula I.Scheme 3

[0199] In some embodiments, the compounds of Formula I are prepared as shown in Scheme 3. Therefore, compounds of Formula A are coupled with suitable boronates of Formula F (wherein Rcand Rdare independently C1-6 alkyl, or are joined to form, together with the B and O atoms therebetween, a 4 to 6 membered saturated or unsaturated ring optionally substituted with one or two C1-3alkyls) under suitable coupling conditions such as Suzuki-Miyaura coupling conditions to provide compounds of Formula G. Subsequent sulfonylation of compounds of Formula G with suitable arylsulfonyl chlorides of Formula H affords compounds of Formula E. Oxidation of compounds of Formula E with suitable oxidizing agents such as meta-chloroperoxybenzoic acid (m-CPBA) to the corresponding sulfoxide followed by substitution with various suitable amines compounds of Formula B affords compounds of Formula I.Scheme 4

[0200] In some embodiments as shown in Scheme 4, compounds of Formula A wherein X1= C (compounds of Formula A- are prepared from intermediate compounds 46 8713541of Formula J. Treatment of compounds of Formula J with a suitable amino alcohol (e.g., aminoethanol) followed by the addition of methanesulfonyl chloride in a suitable base such as triethylamine (Et3N) provides the compounds of Formula A-1.Scheme 5

[0201] In some embodiments shown in Scheme 5, compounds of Formula A wherein X1= N (compounds of Formula A-2) are prepared from the commercially available 4-amino-2-(methylthio)-5-pyrimidinecarboxaldehyde (Compound K). Condensation of K with substituted triethyl phosphonoacetates under basic conditions gives intermediate compounds of Formula L. Base-mediated cyclisation of compounds of Formula L provides the intermediate compounds of Formula M. Selective bromination of compounds of Formula M provides compounds of Formula N, which is then followed by chlorination with suitable chlorinating agents such as POCl3 to give intermediate compounds of Formula O. Treatment of compounds of Formula O with a suitable hydrazines in a suitable solvent such as ethanol gives hydrazide compounds of Formula P which are then subjected to cyclization reaction conditions such as in the presence of formic acid (Z=H) to afford the tricyclic intermediate compounds of Formula A-2.Scheme 6

[0202] In some embodiments as shown in Scheme 6, compounds of Formula C (wherein Raand Rbare independently C1-6or are joined to form, together with the B 47 8713541and O atoms therebetween, a 4 to 6 membered saturated or unsaturated ring optionally substituted with one or two C1-3alkyls) are prepared by coupling aryl sulfonyl compounds of Formula Q with aniline compounds of Formula R (wherein Y' and Y'' are each independently halogen, such as Cl or Br) in the presence of a suitable base such as pyridine. Subsequent borylation of compounds of Formula R under standard borylation conditions such as Miyaura borylation condition using (pinacolato)diboron in basic conditions with a suitable catalyst such as PdCl2(dppf) (([1,1' bis(diphenylphosphino)ferrocene]palladium(II) dichloride) provides the compounds of Formula C.Scheme 7

[0203] In some embodiments as shown in Scheme 7, compounds of Formula C (wherein Raand Rbare independently C1-6alkyl, or are joined to form, together with the B and O atoms therebetween, a 4 to 6 membered saturated or unsaturated ring optionally substituted with one or two C1-3alkyls) are prepared by coupling aryl sulfonyl compounds of Formula Q with anilino boronic ester or acid compounds of Formula T in the presence of a suitable base such as pyridine to provide the compounds of Formula C. Compounds of Formula T are prepared by borylation of compounds of Formula U under borylation conditions such as Miyaura borylation condition using (pinacolato)diboron in basic conditions with a suitable catalyst such as PdCl2(dppf).

[0204] Generally, the reactions described above are performed in a suitable inert organic solvent and at temperatures and for times that will optimize the yield of the desired compounds. Examples of suitable inert organic solvents include, but are not limited to, 2- 48 8713541propanol, dimethylformamide (DMF), 1,4-dioxane, methylene chloride, chloroform, tetrahydrofuran (THF), toluene, and the like.

[0205] Salts of the compounds of the application are generally formed by dissolving the neutral compound in an inert organic solvent and adding either the desired acid or base and isolating the resulting salt by either filtration or other known means.

[0206] The formation of a desired compound salt is achieved using standard techniques. For example, the neutral compound is treated with an acid or base in a suitable solvent and the formed salt is isolated by filtration, extraction or any other suitable method.

[0207] The formation of solvates will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art. Examples of suitable solvents are ethanol, water and the like. When water is the solvent, the molecule is referred to as a “hydrate”. The formation of solvates of the compounds of the application will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art.

[0208] Prodrugs of the compounds of the present application may be, for example, conventional esters formed with available hydroxy, thiol, amino or carboxyl groups. For example, available hydroxy or amino groups may be acylated using an activated acid in the presence of a base, and optionally, in inert solvent (e.g. an acid chloride in pyridine).

[0209] Throughout the processes described herein it is to be understood that, where appropriate, suitable protecting groups will be added to, and subsequently removed from, the various reactants and intermediates in a manner that will be readily understood by one skilled in the art. Conventional procedures for using such protecting groups as well as examples of suitable protecting groups are described, for example, in “Protective Groups in Organic Synthesis”, T.W. Green, P.G.M. Wuts, Wiley-Interscience, New York, (1999). It is also to be understood that a transformation of a group or substituent into another group or substituent by chemical manipulation can be conducted on any intermediate or final product on the synthetic path toward the product, in which the possible type of 49 8713541transformation is limited only by inherent incompatibility of other functionalities carried by the molecule at that stage to the conditions or reagents employed in the transformation. Such inherent incompatibilities, and ways to circumvent them by carrying out appropriate transformations and synthetic steps in a suitable order, will be readily understood to one skilled in the art. Examples of transformations are given herein, and it is to be understood that the described transformations are not limited only to the generic groups or substituents for which the transformations are exemplified. References and descriptions of other suitable transformations are given in “Comprehensive Organic Transformations – A Guide to Functional Group Preparations” R.C. Larock, VHC Publishers, Inc. (1989). References and descriptions of other suitable reactions are described in textbooks of organic chemistry, for example, “Advanced Organic Chemistry”, March, 4th ed. McGraw Hill (1992) or, “Organic Synthesis”, Smith, McGraw Hill, (1994). Techniques for purification of intermediates and final products include, for example, straight and reversed phase chromatography on column or rotating plate, recrystallisation, distillation and liquid-liquid or solid-liquid extraction, which will be readily understood by one skilled in the art.

[0210] The products of the processes of the application may be isolated according to known methods, for example, the compounds may be isolated by evaporation of the solvent, by filtration, centrifugation, chromatography or other suitable method.

[0211] One skilled in the art will recognize that where a reaction step of the present application is carried out in a variety of solvents or solvent systems, said reaction step may also be carried out in a mixture of the suitable solvents or solvent systems EXAMPLES

[0212] The following non-limiting examples are illustrative of the present application: A. Synthesis and Characterization of Exemplary Compounds of the Application GENERAL METHODS General Method MB (Miyaura borylation)

[0213] A degassed 1,4-dioxane mixture of aryl halide (1.0 equiv), B2pin2 (1.3 equiv), KOAc (3.5 equiv) and typically PdCl2(dppf)*CH2Cl2 or PdCl2(dppf) (0.1 equiv) were heated sealed under Ar in a microwave reactor (typically, 90-100 oC) or oil bath (typically, 100-110 ºC). The crude mixture was then most often used directly in the following Suzuki- Miyaura cross coupling step as a crude without further purification. 50 8713541General Method SMC (Suzuki Miyaura Cross-coupling)

[0214] A vial equipped with a stirring bar, filled with Ar or N2, was charged with aryl boronic acid or aryl boronic ester (typically 1-1.5 equiv, in most instances aryl boronic esters were used as crude mixtures in 1,4-dioxane), base (Cs2CO3typically 3 equiv), aryl halide (typically 1 equiv), catalyst / ligand (most often one of: PdCl2(dppf)*CH2Cl2or PdCl2(dppf); typically, 0.1 equiv). The vial was sealed, H2O and organic solvent or a mixture of organic solvents (DME or 1,4-dioxane) were added. The reaction mixture was degassed with Ar or N2by repeated evacuation and refill with the inert gas and then heated, sealed in a microwave reactor or an oil bath for the time specified. After the reaction was deemed complete by LCMS analysis, the mixture was concentrated under reduced pressure, deposited on a plug of Celite® or a SiO2 samplet and purified by flash chromatography (typically using SiO₂ InnoFlash® cartridge or SiO₂ Biotage® cartridge or SiO₂ RediSep®Rf cartridge and hexanes-EtOAc or CH2Cl2–MeOH or CH2Cl2–MeOH-NH3or CH2Cl2–MeOH- conc aq NH3(89:9:1 v / v / v) in CH2Cl2) or preparative HPLC (typically using Biotage® SNAP KP-C₁₈-HS cartridge or RedisSep®Rբ C₁₈ and MeOH in H₂O + 0.05 % TFA or MeCN in H2O + 0.1 % formic acid), optionally followed by a filtration through a Waters PoraPak™ CX column or an Isolute™ CSX-2 column, rinsing with MeOH and eluting the desired material with 2 M NH₃ in MeOH. General Method NS (N-sulfonylation)

[0215] A solution of substituted 3-bromoaniline (1 equiv) and anh pyridine (typically 1.5-2.0 equiv) in CH₂Cl₂ (typically 0.06-0.19 M) was treated with one portion of solid aryl- sulfonyl chloride (typically, 1 equiv) at 0 ºC. The reaction was allowed slowly to reach rt and stirring was continued overnight. The reaction mixture was then concentrated under reduced pressure onto Celite® or alternatively washed by extraction with H₂O, the organic extracts were concentrated under reduced pressure and after depositing on Biotage® samplet or Celite®, purified by flash chromatography on silica gel using one of the cartridges: InnoFlash®, Biotage®, RediSep®Rf. General Method MO (mCPBA mediated oxidation)

[0216] To a suspension of methylthioheteroaryl (1 equiv) in CH2Cl2 was added mCPBA (mCPBA) (typically 1.1-2.2 equiv., technical grade, <77 %) either as a solid or in a small amount of CH2Cl2at 0 °C. The reaction was then allowed to slowly warm to rt and then was stirred at rt until completion. The resultant mixture of the sulfoxide and sulfone intermediates was either washed by with saturated (satd) aq NaHCO3followed 51 8713541by concentration of the organic layer or simply concentrated under reduced pressure and used crude without a workup or further purification. INTERMEDIATES Intermediate 1.6-bromo-2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine Step 1.6-bromo-7-chloro-2-(methylthio)pyrido[2,3-d]pyrimidine

[0217] To 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (5 g, 18.4 mmol) was added POCl3(100 mL), and the mixture was heated at 110 °C overnight. After completion, the reaction mixture was concentrated to 5 mL, and carefully poured into ice- cold H2O. After being stirred for 15 min, the formed solid was filtered, washed with distilled H2O and dried under vacuum to afford the 6-bromo-7-chloro-2-(methylthio)pyrido[2,3- d]pyrimidine as a white solid.(4 g, 75 %). MS (ESI) m / z [M+H]+289.0 / 292.0. Step 2.6-bromo-7-hydrazineyl-2-(methylthio)pyrido[2,3-d]pyrimidine.

[0218] To a suspension of 6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7(8H)-one (4.0 g, 13.8 mmol) in EtOH (40 mL), hydrazine hydrate (20 mL) was added and the reaction mixture was stirred at 80 °C for 2 h. After completion, the reaction mixture was cooled to force precipitation. The solid was filtered, washed with EtOH and dried under reduced pressure to afford 6-bromo-7-hydrazineyl-2-(methylthio)pyrido[2,3-d]pyrimidine as a white solid (3.0 g, 74 %).1H NMR (400 MHz, DMSO-d6): δ 9.22 (br.s., 1H), 8.82 (s, 1H), 8.34 (s, 1H), 4.89 (br.s., 2H). The signal corresponding to Me group is expected to be buried under the solvent peak. Step 3.6-bromo-2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine 52 8713541

[0219] To a 6-bromo-7-hydrazineyl-2-(methylthio)pyrido[2,3-d]pyrimidine (3.0 g, 10 mmol) was added HCO2H (30 mL) and the reaction mixture was stirred at 100 °C for 3 h. After completion, the reaction mixture was poured into the crushed ice, formed solid was filtered, washed with H2O and dried under vacuum to afford 6-bromo-2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine as a greyish white solid (1.4 g, 45 %). MS (ESI) m / z [M+H]+296.1 / 298.0.1H NMR (400 MHz, DMSO-d6): δ 10.00 (s, 1H), 9.23 (s, 1H), 8.24 (s, 1H), 2.70 (s, 3H). Intermediate 2: N-(3-bromo-2,4-difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide

[0220] To a stirred solution of 5-chloro-2-methoxypyridine-3-sulfonyl chloride (20 g, 83 mmol) in pyridine (200 mL) was added 3-bromo-2,4-difluoroaniline (15.5 g, 74.3 mmol) and the resulting reaction mixture was stirred at rt for 1 h. After completion, the reaction mixture was quenched with 2 M aq HCl (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 material was triturated with 2 % EtOAc in hexanes to afford 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 / 414.8.1H 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). Intermediate 3: 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl)-2-methoxypyridine-3-sulfonamide and (5-((5-chloro-2-methoxypyridine)-3- sulfonamido)-2-fluorophenyl)boronic acid

[0221] To a stirred solution of 5-chloro-2-methoxypyridine-3-sulfonyl chloride (4.0 g, 17 mmol) in pyridine (40 mL) was added 4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)aniline (4.0 g, 17 mmol) and the resulting reaction mixture was stirred at rt for 1 h. After completion, the reaction was directly loaded onto silica gel and 53 8713541purified by column chromatography using MeOH in CH2Cl2. Trituration with CH2Cl2and Et2O afforded a mixture of 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan- 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. Intermediate 4: (3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid

[0222] To a cooled solution of 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)aniline (2.5 g, 10.5 mmol) in anhydrous (anh) pyridine (30 mL) was added 5-chloro-2- methoxypyridine-3-sulfonyl chloride (2.5 g, 10.3 mmol) at 0 °C and the reaction mixture was stirred at rt for 16 h. After completion, the reaction mixture was directly loaded onto a silica gel column and purified using MeOH in CH2Cl2to afford (3-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid as a brown solid (2.8 g, 74 %). MS (ESI) m / z [M-H]- 359.3. Intermediate 5: 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

[0223] A solution of 3-bromo-2,4-difluoroaniline (15 g, 72.1 mmol), Pd(dppf)Cl2(5.27 g, 7.21 mmol) and B2pin2(27.4 g, 108.1 mmol), KOAc (21.2 g, 216.3 mmol) in 1,4- dioxane (275 mL) under N2was stirred at 100 °C for 16 h. After completion, the solid was removed by filtration and rinsed with CH2Cl2(150 mL). The filtrate was concentrated under reduced pressure and purified by column chromatography using EtOAc in hexane to afford 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline as a white solid (10 g, 54%). MS (ESI) m / z [M+H]+256.2 and 174.0 (the corresponding boronic acid).1H 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). 54 8713541Intermediate 6. 6-bromo-2-(methylthio)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3- d]g, 14 mmol), 2-aminoethan-1-ol (4.2 g, 69 mmol) was added and the reaction mixture was stirred at 70°C for 4 h. The reaction mixture was then cooled, and cold H2O was added. A solid was filtered, washed with to afford 2-((6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7- yl)amino)ethan-1-ol as a white solid (3.0 g, 69 %). MS (ESI) m / z [M+H]⁺ 315.0 / 316.9. To a solution of 2-((6-bromo-2-(methylthio)pyrido[2,3-d]pyrimidin-7-yl)amino)ethan-1-ol (3.0 g, 9.5 mmol) in dichloromethane (72 mL), triethylamine (4.0 mL, 29 mmol) and MeSO2Cl (1.47 mL, 19.0 mmol) were added at rt, and the reaction mixture was stirred at rt for 3 h. The reaction mixture was then poured into the crushed ice. A solid was filtered, washed with H2O, triturated with MeOH, CH₂Cl₂ and dried under vacuum to afford 6-bromo-2- (methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine as a light green solid (0.75 g, 26 %).1H NMR (400 MHz, DMSO) δ 9.06 (s, 1H), 8.80 (s, 1H), 4.73 – 4.66 (m, 2H), 4.20 – 4.13 (m, 2H), 2.64 (s, 3H). MS (ESI) m / z [M+H]⁺296.9 / 299.0. Intermediate 7. 6-bromo-9-methyl-2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidine.

[0225] A mixture of 6-bromo-7-hydrazineyl-2-(methylthio)pyrido[2,3-d]pyrimidine (2.5 g, 8.7 mmol) and CH3CO2H (10 mL) was stirred at 100 °C for 3 h. The reaction mixture was then was poured into crushed ice. The solid was filtered, washed with H2O, dried and purified by column chromatography using neutral aluminum oxide and 5 % MeOH in DCM to afford 6-bromo-9-methyl-2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine as an orange solid (1.2 g, 44 %). MS (ESI) m / z [M+H]⁺310.0 / 312.0.1H NMR (400 MHz, DMSO) δ 9.17 (s, 1H), 8.16 (s, 1H), 3.10 (s, 3H), 2.69 (s, 3H). EXEMPLARY COMPOUNDS OF THE APPLICATION 55 8713541Example 1: 5-chloro-N-(4-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-1)Step 1: 6-bromo-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine

[0226] To a cold (0 ºC) suspension of 6-bromo-2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (200 mg, 0.67 mmol) in CH₂Cl₂ (60 mL) was added mCPBA (175 mg, 0.71 mmol, > 70 %) as a solid allowing slowly to warm to rt for the total time of 19.6 h. The resulting pale-yellow suspension was concentrated to dryness and use crude in the following step. MS (ESI) m / z [M+H]⁺ 312.07 | 314.20 Step 2: 6-bromo-N-methyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine

[0227] 6-bromo-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (70 mg, 0.23 mmol) was suspended in i-PrOH (12mL) and was treated with MeNH2(40wt % in H2O, 1.2 mL, 13.5 mmol) at rt. Stirred at rt for 1 d 21 h then concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂ using MeOH in CH₂Cl₂ to afford 6-bromo-N-methyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-amine a white solid. MS (ESI) m / z [M+H]⁺ 279.15 / 281.22 Step 3: 5-chloro-N-(4-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-1)

[0228] Following the General Method SMC using the entire material (6-bromo-N- methyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine) from the previous step, Cs2CO3(220 mg, 0.67 mmol), Pd(dppf)Cl2*CH2Cl2(18.20 mg, 0.022 mmol) crude 5-chloro- N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-methoxypyridine-3- sulfonamide (4.6 mL, 0.24 mmol, 0.053 M in 1,4-dioxane) and H2O (2.3 mL); by heating at 100 °C for 2 h. Purified by flash chromatography (SiO₂, using MeOH in CH₂Cl2) and then preparative HPLC (C₁₈, MeCN in H₂O + % HCO₂H) to afford after freeze drying 5- 56 8713541chloro-N-(4-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide as a beige solid (20 mg, 16 % yield). MS (ESI) m / z [M+H]⁺ 515.35. 1H NMR (500MHz, DMSO-d6) δ = 10.61 (br. S., 1H), 9.73 (s, 1H), 9.45 (s, 0.5 H), 9.10 (s, 1H), 9.01 (s, 0.5 H), 8.46 (d, J=2.4 Hz, 1H), 8.24 (d, J=4.5 Hz, 1H), 8.19 - 8.10 (m, 1H), 7.79 - 7.66 (m, 2H), 7.32 - 7.24 (m, 1H), 7.21 - 7.13 (m, 1H), 3.99 (s, 3H), 3.06 - 2.94 (m, 3H). Example 2: trans-5-chloro-N-(3-(2((-4-(dimethylamino)cyclohexyl)amino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3- sulfonamide (trans I-2.HCO2H)Step 1: trans-N1-(6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)-N4,N4- dimethylcyclohexane-1,4-diamine

[0229] trans-N1,N1-dimethylcyclohexane-1,4-diamine, 2HCl (72 mg, 0.34 mmol), 6-bromo-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (70 mg, 0.224 mmol) and K2CO3 (124 mg, 0.90 mmol) were suspended with sonication in DMF (6 mL). The reaction mixture was stirred at rt for 1 d 22 h. The reaction mixture was later concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO2 using MeOH in CH₂Cl₂) to afford trans-N1-(6-bromo- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)-N4,N4-dimethylcyclohexane-1,4- diamine as a beige solid (88 mg, 67 % based on the purity of 67 %). MS (ESI) m / z [M+H]⁺ 390.37|392.38 Step 2: trans-5-chloro-N-(3-(2-((-4-(dimethylamino)cyclohexyl)amino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3- sulfonamide (trans I-2.HCO2H) 57 8713541

[0230] Prepared following the General Method SMC using the entire solid from the previous step, trans-N1-(6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)- N4,N4-dimethylcyclohexane-1,4-diamine, 88 mg), Pd(dppf)Cl2*CH2Cl2(13 mg, 0.016 mmol), Cs2CO3(146 mg, 0.45 mmol), H2O (1.7 mL) and crude 5-chloro-N-(4-fluoro-3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-methoxypyridine-3-sulfonamide (3.4 mL, 0.18 mmol, in dioxane, 0.053 M); by heating at 95 oC for 3 h. Purified by flash chromatography (SiO₂ using MeOH in CH₂Cl₂) and then preparative HPLC (C₁₈, MeCN in H₂O + 0.1 % HCO₂H) to afford after freeze drying 5-chloro-N-(3-(2-(( trans-4- (dimethylamino)cyclohexyl)amino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2-methoxypyridine-3-sulfonamide*HCO2H an off white solid (25.0 mg, 23 yld based on the purity of 95 %). MS (ESI) m / z [M+H]⁺ 626.58. 1H NMR (500 MHz, DMSO- d6) δ ppm 9.72 - 9.78 (m, 0.54 H), 9.44 (s, 0.34 H), 9.06 (s, 0.38 H), 8.98 - 9.03 (m, 0.61 H), 8.44 (d, J=2.45 Hz, 1 H), 8-18-8.40 (m, 2 H), 8.14 - 8.19 (m, 1 H), 7.70 - 7.72 (m, 1 H), 7.65 - 7.69 (m, 1 H), 7.24 - 7.29 (m, 1 H), 7.15 - 7.20 (m, 1 H), 3.98 (s, 3 H), 2.41 - 2.47 (m, 6 H), 1.88 - 2.14 (m, 4 H), 1.31 - 1.59 (m, 4 H). Example 3: N-(3-(2-amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-3)Step 1: 6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine

[0231] To a suspension of 6-bromo-2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (70 mg, 0.224 mmol) in i-PrOH (12 mL) was added aq NH4OH (0.618 mL, 4.93 mmol) at rt for 1 d 21.5 h. The reaction mixture was stirred at rt for 2 d, before being concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO, using MeOH in CH₂Cl₂)to afford 6- bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine as a pale yellow solid (79 mg, 84 % yld based on purity of 63 %). MS [M+H]⁺ 265.23|267.18 58 8713541Step 2: N-(3-(2-amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)- 5-chloro-2-methoxypyridine-3-sulfonamide (I-3)

[0232] Prepared following General Method SMC using the entire solid from the previous step (6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine, 79 mg, 0.19 mmol), Pd(dppf)Cl2*CH2Cl2 (13 mg, 0.016 mmol), Cs2CO3 (146 mg, 0.45 mmol), H2O (1.7 mL) and crude 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)phenyl)-2-methoxypyridine-3-sulfonamide (3.4 mL, 0.18 mmol, 0.053 M in 1,4-dioxane). The mixture was degassed and then heated sealed under At 95 ºC for 3 h. Purified by flash chromatography (SiO₂, using MeOH in CH₂Cl₂) followed by preparative HPLC (C₁₈, MeCN in H₂O + 0.1 % HCO₂H) eluting, pooled fractions: were subsequently filtered through a SCX-2500 mg column, rinsing with MeOH and eluting with 2 M NH₃ in MeOH to afford N-(3-(2-amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-5-chloro- 2-methoxypyridine-3-sulfonamide as a white solid (1 mg, 1 % based on the purity of 97 %). MS (ESI) m / z [M+H]⁺ 501.36. 1H NMR (500 MHz, DMSO-d6) δ ppm 9.49 (s, 1 H), 9.01 (s, 1 H), 8.41 (d, J=2.45 Hz, 1 H), 8.14 (d, J=2.45 Hz, 1 H), 7.66 - 7.76 (m, 3 H), 7.62 (br. S., 1 H), 7.20 - 7.29 (m, 1 H), 7.09 - 7.18 (m, 1 H), 3.96 (s, 3 H). Example 4: 5-chloro-N-(4-fluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-4)Step 1: 6-bromo-N-isopropyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine

[0233] 6-bromo-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (105 mg, 0.34 mmol) was suspended in i-PrOH (12mL) and treated with i-PtNH2 (0.87 mL, 10 mmol) at rt. The suspension was stirred at rt for 2 d 21 h. The solid material was collected by filtration to afford 6-bromo-N-isopropyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-amine as a pale pink solid (105.0 mg, quant). MS (ESI) m / z [M+H]⁺ 307.24 | 309.25 59 8713541Step 2: 5-chloro-N-(4-fluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-4)

[0234] Prepared following General Method SMC using 6-bromo-N-isopropyl- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine (50 mg, 0.16 mmol), Pd(dppf)Cl2*CH2Cl2(13 mg, 0.016 mmol), Cs2CO3(133 mg, 0.41 mmol), H2O (1.88 mL) and crude 5-chloro-N-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2- methoxypyridine-3-sulfonamide (3.8 mL, 0.19 mmol in 1,4-dioxane); by heating at 90 °C for 2 h. Purified by flash chromatography (SiO₂ using MeOH in CH₂Cl₂) and then preparative HPLC (C₁₈, MeCN in H₂O + 0.1 % HCO₂H) to afford 5-chloro-N-(4-fluoro-3-(2- (isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide as a beige solid (20.0 mg , 23 % yield based on purity of 97 %). MS (ESI) m / z [M+H]⁺543.49. 1H NMR (500 MHz, DMSO-d6) δ ppm 9.69 (s, 0.6 H), 9.46 (br. S., 0.3 H), 8.92 - 9.13 (m, 1 H), 8.46 (d, J=2.57 Hz, 1 H), 8.05 - 8.23 (m, 2 H), 7.65 - 7.74 (m, 2 H), 7.26 - 7.32 (m, 1 H), 7.15 - 7.23 (m, 1 H), 4.22 - 4.39 (m, 1 H), 3.99 (s, 3 H), 1.17 - 1.29 (m, 6 H). Example 5: 5-chloro-N-(2-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-5)

[0235] Prepared by General Method SMC using 6-bromo-N-methyl- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine (60 mg, 0.21 mmol), Pd(dppf)Cl2*CH2Cl2 (18 mg, 0.021 mmol), Cs2CO3 (175 mg, 0.54 mmol) in H2O (2.0 mL) and crude 5-chloro-N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2- methoxypyridine-3-sulfonamide (0.052 M in 1,4-dioxane, 4.1 mL, 0.25 mmol,); by heating sealed in an oil bath at 90 °C for 2 h. Purified by flash chromatography (SiO₂ using MeOH in CH₂Cl₂), repurified by preparative HPLC (C₁₈, MeCN in H₂O + 0.1 % HCO₂H) to afford 5-chloro-N-(2-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide as a white solid (34.5 mg , 31 % yield). MS (ESI) m / z [M+H]⁺515.42. 1H NMR (500 MHz, DMSO-d6) δ ppm 10.17 - 10.97 (m, 1 H), 9.71 (s, 0.7 H), 9.48 (s, 0.3 H), 8.92 - 9.14 (m, 1 H), 8.46 (d, J=2.45 Hz, 1 H), 8.19 (d, 60 8713541J=4.77 Hz, 1 H), 8.09 (d, J=2.57 Hz, 1 H), 7.65 (s, 1 H), 7.59 (br. S., 1 H), 7.33 - 7.39 (m, 1 H), 7.23 - 7.30 (m, 1 H), 3.89 (s, 3 H), 2.94 - 3.04 (m, 3 H). Example 6: 5-chloro-N-(2,4-difluoro-3-(2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (I-6)Step 1: 2,4-difluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)aniline

[0236] Prepared by General Method SMC using 2,4-difluoro-3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)aniline (323 mg, 1.266 mmol), 6-bromo-2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (150 mg, 0.51 mmol), Chloro(2- dicyclohexylphosphino-2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2- yl)palladium(II) (40 mg, 0.051 mmol) and K3PO4(376 mg, 1.8 mmol) in H2O (1 mL) and 1,4- dioxane (10 mL); by heating sealed in a mw vial at 70 °C for 4 d. The reaction mixture was cooled to rt, charged with K3PO4(376 mg, 1.8 mmol), Chloro(2-dicyclohexylphosphino- 2',4',6'-tri-i-propyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (40 mg, 0.051 mmol), 2,4-difluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (323 mg, 1.3 mmol) again. The mixture was degassed with Ar and heated in a mw reactor at 65 °C for 6 h. Purified by flash chromatography (SiO₂ using MeOH in CH₂Cl₂) to afford 2,4-difluoro- 3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)aniline as an orange solid (128.0 mg, 71 % yield). MS (ESI) m / z [M+H]⁺ 345.30. Step 2: 5-chloro-N-(2,4-difluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide. 61 8713541

[0237] Prepared by General Method NS using 2,4-difluoro-3-(2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)aniline (128 mg, 0.37 mmol), 5-chloro-2- methoxypyridine-3-sulfonyl chloride (99 mg, 0.41 mmol), pyridine (0.15 mL, 1.9 mmol) in CH₂Cl₂ (18 mL). Purified by flash chromatography (SiO₂ using MeOH in CH₂Cl₂) to afford 5-chloro-N-(2,4-difluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide as a pale yellow solid (151.0 mg , 74 % yield) MS (ESI) m / z [M+H]⁺ 550.40. Step 3: 5-chloro-N-(2,4-difluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide

[0238] To a cold (0 ºC) suspension of 5-chloro-N-(2,4-difluoro-3-(2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (151 mg, 0.27 mmol) in CH₂Cl₂ (12 mL) was added mCPBA (71 mg, 0.29 mmol, >70 %) as a solid and the reaction was allowed slowly to warm to rt for 1.5 h.

[0239] Assuming complete conversion, the material, a 5:1 mixture of 5-chloro-N- (2,4-difluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide and 5-chloro-N-(2,4-difluoro-3-(2- (methylsulfonyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide was concentrated to dryness and used directly in the following step. MS (ESI) m / z [M+H]⁺ 566.32 and 582.29 (the corresponding sulfone). Step 4. 5-chloro-N-(2,4-difluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-6)

[0240] 5-chloro-N-(2,4-difluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- 62 8713541sulfonamide (51.6 mg, 0.091 mmol) was suspended in i-PrOH (10 mL) and treated with MetNH2(40 wt % in H2O, 0.32 mL, 3.6 mmol) at rt. The suspension was stirred at rt for 3 d 21 h. The reaction mixture was concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂ using aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(2,4-difluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as an off-white solid (21.0 mg, 43 % yield). 1H NMR (500 MHz, DMSO-d6) δ ppm 10.26 - 10.71 (m, 1 H), 9.71 (s, 0.7 H), 9.48 (s, 0.3 H), 8.95 - 9.11 (m, 1 H), 8.46 (d, J=2.32 Hz, 1 H), 8.12 - 8.30(m, 1 H), 8.07 (d, J=2.57 Hz, 1 H), 7.70 (s, 1 H), 7.38 - 7.50 (m, 1 H), 7.11 - 7.30 (m, 1 H), 3.89 (s, 3 H), 2.91 - 3.05 (m, 3 H). LC-MS calcd. For [C₂₁H₁₅ClF₂N₈O₃S + H]⁺ 533.06; found MS (ESI) m / z [M+H]⁺ 533.35 Example 7: 5-chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (I-7)

[0241] 5-chloro-N-(2,4-difluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (52 mg, 0.091 mmol) was suspended in i-PrOH (10 mL) and treated with EtNH2 (66-72% in H2O, 0.29 mL, 3.6 mmol) at rt. The suspension was stirred at rt for 3 d 21 h, then concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂, using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-2-methoxypyridine-3-sulfonamide as an off white solid (26 mg, 52 % yield). MS (ESI) m / z [M+H]⁺ 547.33. 1H NMR (500 MHz, DMSO-d6) δ ppm 10.47 (br. S., 1 H), 9.43 - 9.71 (m, 1 H), 8.96 - 9.07 (m, 1 H), 8.47 (d, J=2.57 Hz, 1 H), 8.18 - 8.36 (m, 1 H), 8.07 (d, J=2.57 Hz, 1 H), 7.68 (s, 1 H), 7.37 - 7.49 (m, 1 H), 7.25 (t, J=8.80 Hz, 1 H), 3.90 (s, 3 H), 3.40 - 3.47 (m, 2 H), 1.14 - 1.27 (m, 3 H). 63 8713541Example 8: 5-chloro-N-(2,4-difluoro-3-(2-(isopropylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (I-8)

[0242] 5-chloro-N-(2,4-difluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (52 mg, 0.091 mmol) was suspended in i-PrOH (10 mL) and treated with Isopropylamine (0.31 mL, 3.6 mmol) at 3 d 21. The reaction mixture was concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(2,4-difluoro- 3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide as an off white solid (20 mg ,39 % yield). MS (ESI) m / z [M+H]⁺ 561.43. 1H NMR (500 MHz, DMSO-d6) δ ppm 10.24 - 10.67 (m, 1 H), 9.67 (s, 0.7 H), 9.44 (s, 0.3 H), 8.96 - 9.09 (m, 1 H), 8.46 (d, J=2.45 Hz, 1 H), 8.16 - 8.31(m, 1 H), 8.07 (d, J=2.57 Hz, 1 H), 7.68 (s, 1 H), 7.38 - 7.47 (m, 1 H), 7.24 (t, J=8.68 Hz, 1 H), 4.20 - 4.39 (m, 1 H), 3.87 - 3.92 (m, 3 H), 1.17 - 1.29 (m, 6 H) Example 9: 5-chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-9)Step 1. 5-chloro-N-(4-fluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide 64 8713541

[0243] Prepared following General Method SMC using (5-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (670 mg, 1.86 mmol), 6- bromo-2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (500 mg, 1.69 mmol), Cs2CO3 (1375 mg, 4.22 mmol) and Pd(dppf)Cl2*CH2Cl2 (138 mg, 0.17 mmol) in H2O (20 mL) and 1,4-dioxane (40 mL); by heating at 100 °C overnight. The reaction mix was diluted with H2O (100 mL) and acidified with 1 M aq HCl (8 mL) to pH ~ 1. The solid product was collected by filtration, rinsed with H2O and later triturated (with sonication) using Et2O to afford 5-chloro-N-(4-fluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin- 6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a tan solid (871 mg, 88 % yield based on purity of 91 %). MS (ESI) m / z [M+H]⁺ 532.20. Step 2: 5-chloro-N-(4-fluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide and 5-chloro-N-(4-fluoro-3-(2- (methylsulfonyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide

[0244] To a cold (0 ºC) suspension of 5-chloro-N-(4-fluoro-3-(2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (871 mg, 1.5 mmol, 91 %) in CH₂Cl₂ (70 mL) was added mCPBA (386 mg, 1.6 mmol, >75 %) as a solid in one portion. After 2.5 h, another portion of mCPBA (538 mg, 2.339 mmol) was added as a solid at rt. After 3.5 h of stirring, the reaction was aged 0 ºC for 16.5 h, and stirring at rt was resumed for 2 d. mCPBA (514 mg, 2.235 mmol) was added again in one portion as a solid at rt. Stirring was continued at rt for 19.5 h at this time the reaction was deemed complete and used as mixture of 5-chloro-N-(4-fluoro-3-(2- (methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- 65 8713541methoxypyridine-3-sulfonamide and 5-chloro-N-(4-fluoro-3-(2-(methylsulfonyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide. The crude mixture was concentrated to dryness and used in the following step without purification. MS (ESI) m / z [M+H]⁺ 548.23 and 564.18. Step 3. 5-chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)- 4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-9)

[0245] Crude 5-chloro-N-(4-fluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (104 mg, 0.190 mmol, mixture with the corresponding sulfone) shaken in i- PrOH (12mL) with EtNH2(66-72% in H2O, 0.58 mL, 7.6 mmol) at rt for 2 h 45 min. The reaction mixture was concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂, using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂ to afford 5-chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide as a pale yellow solid (22 mg , 21 % yield based on purity of 98 %). MS (ESI) m / z [M+H]⁺ 529.34. 1H NMR (500 MHz, DMSO-d6) δ ppm 10.34 - 10.81 (m, 1 H), 9.64 - 9.74 (m, 0.6 H), 9.38 - 9.55 (m, 0.3 H), 8.93 - 9.09 (m, 1 H), 8.40 - 8.49 (m, 1 H), 8.24 - 8.31 (m, 0.7 H), 8.18 - 8.23 (m, 0.3 H), 8.13 - 8.18 (m, 1 H), 7.65 - 7.73 (m, 2 H), 7.24 - 7.32 (m, 1 H), 7.13 - 7.22 (m, 1 H), 3.98 (s, 3 H), 3.50-3.75 (m, 2H), 1.09 - 1.26 (m, 3 H). Example 10: 5-chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-10)

[0246] Crude 5-chloro-N-(4-fluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (104 mg, 0.190 mmol, mixture with the corresponding sulfone) shaken in i- PrOH (12mL) with c-PrNH2 (0.523 mL, 7.59 mmol) at rt for 2.7 h. The reaction mixture was concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO. Using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- 66 8713541fluorophenyl)-2-methoxypyridine-3-sulfonamide as a light yellow solid (31.0 mg, 29 % yield based on purity of 96 %). MS (ESI) m / z [M+H]⁺ 541.31. 1H NMR 1H NMR (500 MHz, DMSO-d6) δ ppm 10.36 - 10.77 (brs, 1 H) 9.35 - 9.77 (m, 1 H) 8.91 - 9.17 (m, 1 H) 8.45 (d, J=2.57 Hz, 2 H) 8.17 (d, J=2.57 Hz,1 H) 7.58 - 7.75 (m, 2 H) 7.25 - 7.32 (m, 1 H) 7.16 - 7.22 (m, 1 H) 3.94 (s, 3 H) 2.84 - 3.10 (m, 1 H) 0.47 - 0.89 (m, 4 H). Example 11: 5-chloro-N-(4-fluoro-3-(2-(oxetan-3-ylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (I-11)

[0247] Crude 5-chloro-N-(4-fluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (104 mg, 0.190 mmol, mixture with the corresponding sulfone) was shaken in i-PrOH (12mL) with 3-Oxetanamine (0.53 mL, 7.6 mmol) at rt for 1.1 h. The reaction mixture was concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(4-fluoro-3-(2-(oxetan-3-ylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a pale yellow solid (20.0 mg , 18 % yield based on purity of 97 %). MS (ESI) m / z [M+H]⁺ 557.33. 1H NMR (500 MHz, DMSO-d6) δ ppm 9.72 - 9.81 (m, 0.7 H), 9.51 (br. S., 0.3 H), 9.07 (s, 1 H), 8.93 - 9.03 (m, 1 H), 8.39 - 8.51 (m, 1 H), 8.13 - 8.21(m, 1 H), 7.73 (s, 1 H), 7.69 (dd, J=6.48, 2.69 Hz, 1 H), 7.25 - 7.32 (m, 1 H), 7.11 - 7.22 (m, 1 H), 5.03 - 5.20 (m, 1 H), 4.93 (t, J=6.66 Hz, 1 H),4.79 - 4.88 (m, 1 H), 4.58 - 4.66 (m, 2 H), 3.98 (s, 3 H). Example 12: 5-chloro-N-(3-(2-(ethylamino)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2- 3-sulfonamide67 8713541Step 1. 5-chloro-N-(4-fluoro-3-(2-(methylthio)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide

[0248] Prepared following General Method SMC using (5-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (194 mg, 0.54 mmol), 6- bromo-2-(methylthio)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidine (152 mg, 0.51 mmol), Cs2CO3 (417 mg, 1.3 mmol) and Pd(dppf)Cl2*CH2Cl2 (42 mg, 0.051 mmol) in H2O (5 mL) and 1,4-dioxane (10 mL); by heating sealed in a mw reactor at 90°C for 1.5 h. Purified by flash chromatography (SiO₂, using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(4-fluoro-3-(2-(methylthio)-8,9- dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide as an orange solid (191.0 mg , 66 % yield based on purity of 95 %). MS (ESI) m / z [M+H]⁺ 533.26. Step 2: A mixture of 5-Chloro-N-(4-fluoro-3-(2-(methylsulfinyl)imidazo[1',2':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide, 5-chloro-N-(4-fluoro-3-(2- (methylsulfonyl)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide and 5-chloro-N-(4-fluoro-3-(2-(methylsulfonyl)-8,9- dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamideStep 3. A mixture 5-Chloro-N-(4-fluoro-3-(2-(methylsulfinyl)imidazo[1',2':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide, 5-chloro-N-(4-fluoro-3-(2- (methylsulfonyl)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide and 5-chloro-N-(4-fluoro-3-(2-(methylsulfonyl)-8,9- 68 8713541dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide

[0249] To a cold (0 ºC) suspension of 5-chloro-N-(4-fluoro-3-(2-(methylthio)-8,9- dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (191 mg, 0.340 mmol) in CH₂Cl₂ (20 mL) was added mCPBA (88 mg, 0.36 mmol, >70 %) as a solid and stirred with cooling allowing slowly to warm to rt for the total time of 3 h. The reaction was then aged at 0 oC for 3 d 19 h. Stirring at rt was resumed and continued for 2.5 h before mCPBA (123 mg, 0.53 mmol,75 %) was added as a solid at rt. After 1 h of stirring the reaction was aged at -20 ºC. For 1 d 17 h.

[0250] After 6.5 h of stirring at rt another portion of mCPBA (117 mg, 0.51 mmol, 75 %) was added in one portion as a solid at rt. Stirring was continued for 19 h. At this time the starting material was consumed and the reaction mixture consisted of 5-chloro-N-(4- fluoro-3-(2-(methylsulfinyl)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide MS (ESI) m / z [M+H]⁺ 547.3, 5-chloro-N-(4-fluoro-3-(2- (methylsulfonyl)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide MS (ESI) m / z [M+H]⁺ 563.2, 5-chloro-N-(4-fluoro-3-(2-(methylsulfonyl)-8,9- dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide MS (ESI) m / z [M+H]⁺ 565.3 in an approximate ratio of 1:1:0.5 respectively as judged by UV DAD.

[0251] The material containing the three different products was concentrated under reduced pressure and used crude in the following step. 69 8713541Step 5. 5-chloro-N-(3-(2-(ethylamino)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-12)

[0252] 50 % of crude mixture of 5-Chloro-N-(4-fluoro-3-(2- (methylsulfinyl)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide, 5-chloro-N-(4-fluoro-3-(2-(methylsulfonyl)imidazo[1',2':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide and 5-chloro-N-(4-fluoro-3-(2- (methylsulfonyl)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide from the mCPBA oxidation step was shaken in i-PrOH (10 mL) with EtNH2 (66-72% in H2O, 0.11 mL, 1.4 mmol) at rt for 2.5 h. The reaction mixture was concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (Sousing CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) isolating two different materials: (1) an off-white solid corresponding to m / z 528 and (2) a light yellow solid corresponding to m / z 530.

[0253] The off white solid (1) (m / z 528) was repurified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(3-(2- (ethylamino)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2- methoxypyridine-3-sulfonamide as a white solid (9.0 mg, 17 % yield, based on the estimated amounts for the two starting materials - 5-Chloro-N-(4-fluoro-3-(2- (methylsulfinyl)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide, 5-chloro-N-(4-fluoro-3-(2-(methylsulfonyl)imidazo[1',2':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide combined). MS (ESI) m / z [M+H]⁺ 528.22. 1H NMR (500 MHz, DMSO-d6) δ = 10.78 - 10.20 (br.s., 1H), 9.07 - 8.94 (m, 1H), 8.47 (d, J = 2.4 Hz, 1H), 8.34 (s, 0.7H), 8.22 (br s, 0.3H), 8.18 - 8.13(m, 1H), 8.12 - 8.05 (m, 0.7H), 8.04 - 7.97 (m, 0.3H), 7.62 - 7.59 (m, 1H), 7.57 (br s, 2H), 7.27 - 7.21 (m, 1H), 7.17 - 7.10 (m, 1H), 3.97 (s, 3H), 1.27 - 1.11 (m, 3H). The CH2's signal is obscured by the solvent peak. Example 13: 5-chloro-N-(3-(2-(ethylamino)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3- d]pyrimidin-6-yl)-4- -2- 3-sulfonamide (I-13)70 8713541

[0254] The light yellow solid isolated from the first column chromatography (m / z 530) described in the last step of the synthesis of 5-chloro-N-(3-(2- (ethylamino)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2- methoxypyridine-3-sulfonamide was repurified by flash chromatography (SiO2, using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂ to afford 5-chloro-N-(3-(2-(ethylamino)- 8,9-dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2- methoxypyridine-3-sulfonamideas a pale yellow solid (6.6 mg, 22 %, based on the estimated amount of the corresponding sulfone in the crude starting material: 5-chloro-N- (4-fluoro-3-(2-(methylsulfonyl)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide). MS (ESI) m / z [M+H]⁺ 530.25. 1H NMR (500 MHz, METHANOL-d4) δ = 8.30 (d, J = 2.6 Hz, 1H), 8.27 - 8.23 (m, 1H), 8.11 (d, J = 2.6 Hz, 1H), 7.31 - 7.22 (m, 2H), 7.17 - 7.12 (m, 1H), 7.05 (t, J = 9.2 Hz, 1H), 4.59 (br s, 1H), 4.30 - 4.13 (m, 2H), 4.07 (s, 3H), 4.03 - 3.95 (m, 2H), 3.53 - 3.41 (m, 2H), 1.26 - 1.20 (m, 3H). Example 14: 5-chloro-N-(2-fluoro-3-(2-(methylthio)-[1,2,4]triazolo[4’,3’:1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-14)Step 1. 5-chloro-N-(2-fluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide

[0255] Prepared following General Method SMC using 6-bromo-2-(methylthio)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidine (200 mg, 0.67 mmol), (3-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (243 mg, 0.67 mmol), Cs2CO3 (550 mg, 1.7 mmol) and Pd(dppf)Cl2 (49 mg, 0.068 mmol) in H2O (7 mL) and 1,4- dioxane (14 mL); by heating at 100 °C for 3.5 d. The reaction mixture was cooled to rt, 71 8713541diluted with H2O to 100 mL, acidified with 1 M aq HCl (3 mL) to pH ~ 1. The newly formed precipitate was collected by filtration. The filter cake was rinsed with H2O. Purified by flash chromatography (SiO₂, using MeOH in EtOAc) to afford 5-chloro-N-(2-fluoro-3-(2- (methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine- 3-sulfonamide as a light yellow solid (58.0 mg, 16 % based on the purity of 96 %). MS (ESI) m / z [M+H]⁺ 532.20. 1H NMR (500 MHz, DMSO-d6) δ = 10.61 - 10.42 (m, 1H, NH), 9.95 (s, 1H), 9.36 (s, 1H), 8.50 (d, J = 2.6 Hz, 1H), 8.12 (d, J = 2.6 Hz, 1H), 7.85 (s, 1H), 7.66 (br t, J = 6.6 Hz, 1H), 7.49 - 7.22 (m, 2H), 3.90 (s, 3H), 2.73 (s, 3H). Step 2. 5-chloro-N-(2-fluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide

[0256] 5-chloro-N-(2-fluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (54 mg, 0.10 mmol) in CH₂Cl₂ (10 mL) was treated with mCPBA (32.5 mg, 0.13 mmol, 70 %) added as a suspension in CH₂Cl₂ (1 mL) at 0 ºC. After brief stirring the cooling bath was removed and stirring was continued at rt for 50 min. The reaction mixture was aged at -20 ºC for 2 d. The reaction mixture was concentrated to remove CH₂Cl₂ leaving 5-chloro-N-(2-fluoro-3-(2- (methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2- methoxypyridine-3-sulfonamide as a pale yellow solid that was used crude in the following step. MS (ESI) m / z [M+H]⁺548.20 Step 3. N-(3-(2-amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2-fluorophenyl)- 5-chloro-2-methoxypyridine-3-sulfonamide (I-14)

[0257] The entire 5-chloro-N-(2-fluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide from the previous step was suspended in i-PrOH (10 mL) and treated with conc aq NH3 (28-30%, 0.30 mL, 4.3 mmol) added in one portion at rt. The reaction was shaken at rt for 2 h and then concentrated under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford N-(3-(2-amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2- 72 8713541fluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide as a white solid (11.0 mg, 22 % yld). MS (ESI) m / z [M+H]⁺ 501.18. 1H NMR (500 MHz, DMSO-d6) δ = 10.48 (s, 1H), 9.51 - 9.31 (m, 1H), 9.03 (s, 1H), 8.49 (d, J = 1.7 Hz, 1H), 8.10 (d, J = 2.4 Hz, 1H), 7.81 - 7.68 (m, 2H), 7.67 - 7.64 (m, 1H), 7.64 - 7.57 (m, 1H), 7.41 - 7.34 (m, 1H), 7.33 - 7.23 (m, 1H), 3.90 (s, 3H). Example 15: 5-chloro-N-(4-fluoro-3-(9-methyl-2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (I-15)Step 1.6-bromo-N,9-dimethyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-amine

[0258] 6-bromo-9-methyl-2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidine (300 mg, 0.97 mmol) in CH₂Cl₂ (10 mL) was treated with mCPBA (310 mg, 1.26 mmol, 70 %) added as a suspension in CH₂Cl₂ at 0 ºC (1mL). After brief stirring the cooling bath was removed and stirring was continued at rt for 1.4 h. The reaction mixture was aged -20 ºC) for overnight storage. The reaction mixture was complete at this time and consisted of 6-bromo-9-methyl-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidine and 6-bromo-9-methyl-2-(methylsulfonyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidine (ratio of 1.5:1 by UV). MS (ESI) m / z [M+H]⁺ 326.04|328.15 and MS (ESI) m / z [M+H]⁺ 342.06|344.09.

[0259] The entire material was concentrated to remove CH₂Cl₂ leaving a pale yellow-orange solid that was suspended in i-PrOH (10 mL) and treated with MetNH2 (40wt % in H2O , 1.0 mL, 12 mmol) added in one portion at rt. The reaction was shaken for 1.8 h. Later the suspension was diluted with H2O and filtered. The collected solid was rinsed with H2O to afford 6-bromo-N,9-dimethyl- triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2- 73 8713541amine as a pale pink solid (249.0 mg , 83 % yield based on purity of 95 %). MS (ESI) m / z [M+H]⁺ 293.09| 295.12 Step 2. 5-chloro-N-(4-fluoro-3-(9-methyl-2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (I-15)

[0260] Prepared following General Method SMC using (5-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (106 mg, 0.29 mmol), Cs2CO3(185 mg, 0.57 mmol), 6-bromo-N,9-dimethyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-amine (70 mg, 0.23 mmol) and Pd(dppf)Cl2(17mg, 0.023 mmol) in 1,4- dioxane (4 mL) and H2O (2 mL); by heating in a mw reactor at 100 °C for 2 h. Purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(4-fluoro-3-(9-methyl-2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a white solid (82.0 mg , 68 % yield). MS (ESI) m / z [M+H]⁺ 529.11. 1H NMR (500 MHz, DMSO-d6) δ = 10.4 (s, 1H), 9.02 - 8.88 (m, 1H), 8.47 (d, J = 2.6 Hz, 1H), 8.17 (d, J = 2.6 Hz, 1H), 8.15 - 8.11 (m, 0.8H), 8.01 - 7.95 (m, 0.2H), 7.67 - 7.62 (m, 1H), 7.59 (s, 1H), 7.31 - 7.24 (m, 1H), 7.22 - 7.13 (m, 1H), 3.99 (s, 3H), 3.19 - 3.04 (m, 3H), 2.98 (br d, J = 4.6 Hz, 3H). Example 16: 5-chloro-N-(2-fluoro-3-(9-methyl-2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (I-16)

[0261] Prepared following General Method SMC using (3-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (82 mg, 0.23 mmol), Cs2CO3 (185 mg, 0.57 mmol), 6-bromo-N,9-dimethyl-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-amine (70 mg, 0.23 mmol) and Pd(dppf)Cl2 (17 mg, 0.023 mmol) in 1,4- dioxane (4 mL) and H2O (2 mL; by heating in a mw reactor at 100 °C for 2 h. Purified by flash chromatography (SiO2 using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford 5-chloro-N-(2-fluoro-3-(9-methyl-2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide as a light orange solid (24.5 mg 74 8713541, 20 % yield based on purity of 97%). MS (ESI) m / z [M+H]⁺ 529.16. 1H NMR (500 MHz, DMSO-d6) δ = 10.50 - 10.39 (m, 1H), 9.03 - 8.87 (m, 1H), 8.49 (d, J = 2.4 Hz, 1H), 8.17 - 8.11 (m, 0.7H), 8.11 - 8.08 (m, 1H), 8.00 - 7.94 (m, 0.2H), 7.60 - 7.54 (m, 1H), 7.54 - 7.49 (m, 1H), 7.38 - 7.33 (m, 1H), 7.30 - 7.24 (m, 1H), 3.92 - 3.88 (m, 3H), 3.15 - 3.03 (m, 3H), 3.01 - 2.94 (m, 3H) Example 17: trans-4-((6-(5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1- carboxamide (trans I-17)Step 1. (1R, 4R)-4-((6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N- methylcyclohexane-1-carboxamide

[0262] Crude 6-bromo-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidine (105.5mg, 0.34 mmol), (1R, 4R)-4-amino-N-methylcyclohexane-1- carboxamideTFA (137 mg, 0.51 mmol) and K2CO3 (173 mg, 1.2 mmol) in NMP (4 mL) were shaken at rt for 3.1 h. The reaction was diluted with H2O (10 mL). After overnight standing at rt, a collected by filtration to afford trans-4-((6-bromo- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1- carboxamideas an off white solid (89 mg , 65 % yield for 2 steps). MS (ESI) m / z [M+H]⁺ 404.28 / 406.23 Step 2. trans-4-((6-(5-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1- carboxamide (I-17)

[0263] Prepared following General Method SMC using (5-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2- boronic acid (51.6 mg, 0.143 mmol), 75 8713541Cs2CO3 (90 mg, 0.275 mmol), (1R, 4R)-4-((6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (44.5 mg, 0.110 mmol) and Pd(dppf)Cl2 (8.0 mg, 0.011 mmol) in 1,4-dioxane (8 mL) and H2O (4 mL); by heating at 100 °C for 2.1 h. Purified by flash chromatography (SiO2, using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂) to afford trans-4-((6-(5-((5-chloro-2-methoxypyridine)-3- sulfonamido)-2-fluorophenyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N- methylcyclohexane-1-carboxamide as a beige solid (36 mg , 50 % yield based on purity of 97 %). MS (ESI) m / z [M+H]⁺ 640.29. 1H NMR (500 MHz, DMSO-d6) δ = 10.58 (m, 1H), 9.79 (s, 0.6H), 9.40 (s, 0.4H), 9.11 - 9.05 (m, 0.4H), 9.03 - 8.96 (m, 0.6H), 8.47 (d, J = 2.3 Hz, 1H), 8.24 (br d, J = 8.1 Hz, 0.6H), 8.18 (d, J = 2.6 Hz, 1.4H), 7.77 - 7.63 (m, 3H), 7.32 - 7.24 (m, 1H), 7.23 - 7.13 (m, 1H), 4.00 (s, 3H), 3.95-3.85 (m, 1H), 2.61 - 2.54 (m, 3H), 2.17 - 1.92 (m, 3H), 1.83 - 1.75 (m, 2H), 1.69 - 1.54 (m, 1H), 1.52 - 1.41 (m, 1H), 1.41 -1.28 (m, 2H). Example 18: trans-4-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1- carboxamide (trans- I-18)

[0264] Prepared following General Method SMC using (3-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (51.6 mg, 0.143 mmol), Cs2CO3 (90 mg, 0.27 mmol), trans-4-((6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide (44.5 mg, 0.11 mmol) and Pd(dppf)Cl2 (8.0 mg, 0.011 mmol) in 1,4-dioxane (4 mL) and H2O (2 mL); by heating in a mw reactor at 100 °C for 2. H. Purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂ to afford trans-4-((6-(3-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1-carboxamide as a white solid (29 mg , 41 % yield). MS (ESI) m / z [M+H]⁺ 640.37.1H NMR (500 MHz, DMSO-d6) δ ppm 10.48 (br. s., 1 H), 9.79 (s, 0.7 H), 9.40 (s, 0.3 H), 8.95 - 9.13 (m, 1 H), 8.48 (d, J=2.69 Hz, 1 H), 8.13 - 8.30 (m,1 H), 8.10 (d, J=2.57 Hz, 1 H), - 7.78 (m, 1 H), 7.63 (s, 2 H), 7.32 - 7.43 (m, 76 87135411 H), 7.17 - 7.31 (m, 1 H), 3.96 - 4.07 (m, 1 H), 3.90 (s, 3H),2.55 - 2.61 (m,3 H), 1.93 - 2.16 (m, 3 H), 1.75 - 1.83 (m, 2 H), 1.57 - 1.71 (m, 1.5 H), 1.43 - 1.53 (m, 0.8 H), 1.27 - 1.40 (m, 2 H). Example 19. trans-4-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide (trans I-19)Step 1. trans-4-((6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2- yl)amino)cyclohexane-1-carboxamide

[0265] Crude 6-bromo-2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidine (105.5mg, 0.39 mmol) trans-4-aminocyclohexane-1-carboxamide hydrochloride (91 mg, 0.51 mmol) and K2CO3 (173 mg, 1.2 mmol) were shaken into NMP (4 mL) at rt for 3.1 h. The reaction was diluted with H2O (10 mL) then concentrated to dryness under reduced pressure, deposited on Celite® and purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂ to afford trans-4-((6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane- 1-carboxamide as a beige solid (48.0 mg , 36 % yield for 2 steps). MS (ESI) m / z [M+H]⁺ 390.15|390.18. Step 2. trans-4-((6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide

[0266] Prepared following General Method SMC using 3-((5-chloro-2- methoxypyridine)-3-sulfonamido)-2-fluorophenyl)boronic acid (58 mg, 0.16 mmol), Cs2CO3(100 mg, 0.31 mmol), trans-4-((6-bromo-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2- yl)amino)cyclohexane-1-carboxamide (48 0.12 mmol) and Pd(dppf)Cl2(9.0 mg, 0.012 77 8713541mmol) in 1,4-dioxane (4 mL) and H2O (2 mL); by heating at 100 °C for 2. h. Purified by flash chromatography (SiO₂ using CH₂Cl₂ / MeOH / conc aq NH₄OH 89 / 10 / 1 in CH₂Cl₂). The collected fractions were concentrated under reduced pressure to 5-10 mL allowing a white precipitate to form. After overnight standing at rt, the white solid was collected by filtration and after rinsing with MeOH afforded trans-4-((6-(3-((5-chloro-2-methoxypyridine)-3- sulfonamido)-2-fluorophenyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2- yl)amino)cyclohexane-1-carboxamide a white solid ( 40 mg, 52 %). MS (ESI) m / z [M+H]⁺ 626.60.1H NMR (500 MHz, DMSO) δ 10.47 (s, 1H), 9.76 (s, 0.6H), 9.41 (s, 0.4H), 9.07 (s, 0.4H), 9.00 (s, 0.6H), 8.48 (s, 1H), 8.23 (d, J = 8.1 Hz, 0.6H), 8.18 (d, J = 8.1 Hz, 0.4H), 8.10 (s, 1H), 7.63 (s, 2H), 7.37 (t, J = 7.5 Hz, 1H), 7.31 – 7.20 (m, 2H), 6.71 (s, 1H), 3.98 (d, J = 8.1 Hz, 1H), 3.90 (s, 3H), 2.15-1.95 (m, 3H), 1.82 (d, J = 12.4 Hz, 2H), 1.60 (q, J = 12.3 Hz, 1H), 1.50-1.30 (m, 3H). Example 20. ((5-Chloro-2-methoxypyridin-3-yl)sulfonyl)(4-fluoro-3-(2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)amide, Potassium (I-1 K)

[0267] A suspension of 5-chloro-N-(4-fluoro-3-(2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (352.7 mg, 0.66 mmol) in EtOH (60 mL)) was sonicated for about 2 min and treated with aq KOH (0.5 M, 1.32 mL, 0.66 mmol) at rt. H2O (60 mL) was added to yield a transparent light tan solution. The excess of EtOH was removed under reduced pressure. The aq solution was freeze-dried to afford ((5-chloro-2-methoxypyridin-3-yl)sulfonyl)(4- fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)amide, potassium as a beige powder (341.0 mg, 91 %). MS (ESI) m / z [M+H]⁺ 515.32. 1H NMR(500 MHz, DMSO-d6) δ 9.69 (s, 0.6H), 9.45 (s, 0.4H), 9.08 (s, 0.3H), 8.99 (s, 0.7H), 8.15 (s, 1.6H), 8.04 (s, 0.4H), 7.96 (s, 1H), 7.56 (s, 1H), 7.15 (d, J = 6.1 Hz, 1H), 6.92 (t, J = 9.5 Hz, 1H), 6.87-6.80 (m, 1H), 3.79 (s, 3H), 3.03 – 2.93 (m, 3H). 19F NMR (471 MHz, DMSO-d6) δ -128.49. Example 21. 5-Chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin- 6-yl)-2-fluorophenyl)-2-methoxypyridine-3- (I-20) 78 8713541

[0268] 5-Chloro-N-(2-fluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (60 mg, 0.11 mmol) in i-PrOH (5 mL) was treated with EtNH2 (66-72% in H2O, 0.27 mL, 3.3 mmol) and shaken at rt for 1.2 h and then stirred at rt overnight. The reaction mixture was concentrated under reduced pressure, deposited on Celite and purified by flash chromatography (SiO2, EtOAc in CH₂Cl2) to afford 5-chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide as a white solid (32.0 mg, 54 % yield based on purity of 98 %).

[0269] MS (ESI) m / z [M+H]⁺ 529.16. 1H NMR(500 MHz, DMSO-d6) δ 10.48 (s, 1H), 9.70 (s, 0.6H), 9.43 (s, 0.4H), 9.09 (s, 0.4H), 9.02 (s, 0.6H), 8.49 (d, J = 2.4 Hz, 1H), 8.33 (t, J = 5.3 Hz, 0.6H), 8.27 – 8.21 (m, 0.4H), 8.11 (d, J = 2.5 Hz, 1H), 7.65 (s, 1H), 7.63 – 7.59 (m, 1H), 7.38 (t, J = 7.0 Hz, 1H), 7.29 (t, J = 7.9 Hz, 1H), 3.90 (s, 3H), 3.58 – 3.40 (m, 2H), 1.29 – 1.15 (m, 3H). Example 22. 5-Chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)-2-fluorophenyl)-2-methoxypyridine-3-sulfonamide (I-21)

[0270] 5-Chloro-N-(2-fluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (60 mg, 0.11 mmol) in i-PrOH (5 mL) was treated with c-PrNH2 (0.27 mL, 3.8 mmol) and shaken at rt for 1.2 h and then stirred at rt overnight. 79 8713541

[0271] The reaction mixture was concentrated under reduced pressure, deposited on Celite and purified by flash chromatography (SiO2, MeOH in CH₂Cl₂) to afford 5-chloro- N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2- fluorophenyl)-2-methoxypyridine-3-sulfonamide as a pale yellow solid (42 mg, 69 % yield based on purity of 97 %). MS (ESI) m / z [M+H]⁺ 541.20. 1H NMR(500 MHz, DMSO-d6) δ 10.47 (s, 1H), 9.67 (s, 0.6H), 9.39 (s, 0.4H), 9.13 (s, 0.4H), 9.02 (s, 0.6H), 8.54 – 8.33 (m, 2H), 8.10 (d, J = 2.5 Hz, 1H), 7.71 – 7.54 (m, 2H), 7.37 (t, J = 7.2 Hz, 1H), 7.28 (t, J = 7.8 Hz, 1H), 3.90 (s, 3H), 3.12 – 2.90 (m, 1H), 0.92 – 0.69 (m, 2H), 0.68 – 0.47 (m, 2H). Example 23.5-Chloro-N-(2-fluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (I-22)

[0272] 5-Chloro-N-(2-fluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (60 mg, 0.11 mmol) in i-PrOH (5 mL) was treated with i-PrNH2 (0.25 mL, 3.0 mmol) and shaken at rt for 1.2 h and stirred at rt overnight. The reaction mixture was concentrated under reduced pressure, deposited on Celite and purified by flash chromatography (SiO2, EtOAc in CH₂Cl2) to afford 5-chloro- N-(2-fluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide as an off white solid (37 mg, 60 % yield based on purity of 97 %)

[0273] MS (ESI) m / z [M+H]⁺ 543.23. 1H NMR(500 MHz, DMSO-d6) δ 10.54 (s, 1H), 9.75 (s, 0.6H), 9.46 (s, 0.4H), 9.15 (s, 0.4H), 9.09 (s, 0.6H), 8.56 (d, J = 2.4 Hz, 1H), 8.32 (d, J = 7.9 Hz, 0.6H), 8.23 (d, J = 7.1 Hz, 0.4H), 8.17 (t, J = 5.2 Hz, 1H), 7.75 – 7.65 (m, 2H), 7.44 (t, J = 7.0 Hz, 1H), 7.35 (t, J = 7.8 Hz, 1H), 4.46 – 4.29 (m, 1H), 3.97 (s, 3H), 1.31 (m, 6H). Example 24. 5-Chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide (I-23) 80 8713541

[0274] 5-Chloro-N-(2,4-difluoro-3-(2-(methylsulfinyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3- sulfonamide (75 mg, 0.13 mmol) in i-PrOH (10 mL) was treated with c-PrNH2 (0.32 mL, 4.6 mmol) and shaken at rt for 20 min and then stirred at rt for 1.8 h. The reaction mixture was concentrated under reduced pressure, deposited on Celite and purified by flash chromatography (SiO₂, MeOH in CH₂Cl2) to afford 5-chloro-N-(3-(2-(cyclopropylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-2-methoxypyridine- 3-sulfonamide as a white solid (13 mg, 18 %). MS (ESI) m / z [M+H]⁺ 559.18.

[0275] 1H NMR(500 MHz, CD3OD) δ 9.70 – 9.52 (m, 1H), 9.10 – 8.83 (m, 1H), 8.32 (d, J = 2.5 Hz, 1H), 8.09 (d, J = 2.5 Hz, 1H), 7.68 (s, 1H), 7.62 (td, J = 8.9, 5.8 Hz, 1H), 7.14 (t, J = 8.9 Hz, 1H), 4.00 (s, 3H), 3.07 – 2.84 (m, 1H), 0.92 (br s, 2H), 0.60 (br s, 2H). Example 25. N-(3-(2-Amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide (I-24)5-Chloro-N-(2,4-difluoro-3-(2-(methylsulfinyl)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide (75 mg, 0.13 mmol) in i-PrOH (10 mL) was treated with NH3 (7 M in MeOH, 0.76 mL, 5.3 mmol) and shaken at rt for 20 min and then stirred at rt for 1.8 h. The reaction mixture was concentrated under reduced pressure, deposited on Celite and purified by flash chromatography (SiO₂, MeOH in CH₂Cl2) to afford N-(3-(2-amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-5-chloro-2-methoxypyridine-3-sulfonamide as a white solid (21 mg, 30 % yield). MS (ESI) m / z [M+H]⁺ 519.16. 1H NMR(500 MHz, DMSO-d6) δ 10.47 (s, 1H), 9.46 81 8713541(s, 1H), 9.02 (s, 1H), 8.49 (d, J = 2.1 Hz, 1H), 8.09 (d, J = 2.5 Hz, 1H), 7.81 (s, 2H), 7.70 (s, 1H), 7.47-7.40 (m, 1H), 7.25 (t, J = 8.6 Hz, 1H), 3.91 (s, 3H). B. Biological Assays GCN2 Enzymatic Assay

[0276] To identify small molecule GCN2 inhibitors, a biochemical GCN2 enzymatic assay was outsourced to Eurofins. This assay was done radiometrically which utilizes full- length, GST-tagged GCN2 (E556G) produced in insect cells. The kinase concentration is 18.5 nM with 70 uM ATP (Km=77 uM) and [g-33P]-ATP in a Tris buffer containing 300 uM of an optimized peptide substrate (RSRSRSRSRSRSRSR). The reaction was initiated by the addition of the Mg / ATP mix. After incubation for 40 minutes at room temperature, the reaction was stopped by the addition of phosphoric acid to a concentration of 0.5%.10 μL of the reaction was then spotted onto a P30 filtermat and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting. The results are shown in Table 1 where IC50’s is reported in the following ranges: A: 0.1- 100 nM; B: 100-1000 nM; C: 1000-10000 nM; D: >10000 nM for the compounds of Formula (I) Table 1: IC50’s(nM) for exemplary compounds of the application for inhibition of GCN2 Compound I.D. GCN2 IC50 (nM) I-1 A trans I-2 A I-3 A I-4 A I-5 A I-6 B I-7 A I-8 B I-9 A I-10 A I-11 A I-12 A I-13 A I-14 A I-15 A 82 8713541Compound I.D. GCN2 IC50(nM) I-16 A trans I-17 A trans I-18 A trans I-19 A I-1 K A I-20 A I-21 A I-22 A I-23 A I-24 A Cell-based phosho- eIF2α Assay:

[0277] To confirm target engagement in cells, an AlphaLISA assay (Perkin Elmer #TGREIR2S10K) was optimized to monitor eIF2α phosphorylation on Serine-51. This event is specifically catalyzed by GCN2 induced with halofuginone (an inhibitor of glutamyl-prolyl tRNA synthetase), borrelidin (an inhibitor of threonyl-tRNA synthetase) or L-asparaginase which activates GCN2 kinase activity by triggering the amino acid starvation response. SKOV3 or U2OS cells (seeded at 40,000 cell per well) are pretreated with exemplary GCN2 inhibitor compounds of the application (1 nM to 1 µM) for 1 hour, stimulated with borrelidin (10 µM) for 1 hour, then lysed and analyzed with the AlphaScreen SureFire kit, which utilizes an antibody based method to quantitatively detect phospho-eIF2α in an HTS format. Tumor Cell Growth Inhibition Assay:

[0278] SKOV3 / OVACR8 cells were seeded into a 384-well plate at 1,000 cells / well in 50ul medium (Alpha-MEM containing 10% FBS, 100 mg / ml Normocin, Invivogen and 50 mg / ml Gentamycin, Invitrogen). Plates were then incubated overnight for the cells to attach. An HP D300 digital dispenser was used to dose cells with ASNase, DMSO or test compounds across a 16-point range of concentrations (high dose of 10uM to low dose of 5nM). Plates were incubated in a humidified 5% CO2 incubator at 37oC. After 3-5 days, plates were removed from the incubator and equilibrated to room temperature. An equal volume of ATPlite assay reagent was then added to each well, and samples processed 83 8713541according to manufacturer’s instructions (Perkin Elmer). Luminescent signals were then measured using an Envision plate reader equipped with a US-Luminescence detector.

[0279] While the present application has been described with reference to what are presently considered to be the preferred examples, it is to be understood that the application is not limited to the disclosed examples. To the contrary, the present application is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

[0280] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term. 84 8713541

Claims

CLAIMS:

1. A compound of Formula I, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof:wherein R1is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two R8a; ---- is a single or a double bond; X1is selected from CR9and N when ---- is a double bond and X1is selected from CR9R9aand NR9bwhen ---- is a single bond; R2is selected from H, C1-4alkyl and C1-4haloalkyl; X2is selected from N and CR10; R3, R4and R5are independently selected from H, halo, CN, C1-6alkyl and C1-6haloalkyl; X3is selected from N and CR11; R6and R7are independently selected from H, halo, CN, C1-6alkyl, C1-6haloalkyl, OC1-6alkyl and OC1-6haloalkyl; R8is selected from H, C1-4alkyl and C1-4haloalkyl; each R8ais independently selected from OR12, NR12R13, C(O)NR12R13, halo, C1-6alkyl, C1-6haloalkyl, C2-6alkenyl, C2-6alkynyl, C3-10cycloalkyl and C3-10heterocycloalkyl, wherein all alkyl, alkenyl, alkynyl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents selected from halo, OR14, NR14R15and C1-6alkyl; R9, R9a, R10and R11are independently selected from H, halo, C1-6alkyl and C1-6haloalkyl; R9bis selected from H, C1-6alkyl and C1-6haloalkyl; R12is selected from H, C1-6alkyl, C1-6haloalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two substituents selected from halo, OH, OC1-4alkyl and OC1-4fluoroalkyl; and R13, R14and R15are independently selected from H, C1-6alkyl and C1-6haloalkyl. 85 87135412. The compound of claim 1, wherein R1is selected from C3-10cycloalkyl and C3-10heteroycloalkyl, each of which is optionally substituted with one or two R8a.

3. The compound of claim 1, wherein R1is selected from H, C1-4alkyl and C1-4fluoroalkyl.

4. The compound of claim 3, wherein R1is selected from H, CH3, CF3, CHF2, CH2CH3, CH2CH2F, CH2CH2CH3, CH(CH3)2, CH(CH3)CH2CH3and CH(CH3)3.

5. The compound of claim 1, wherein R1is C3-10cycloalkyl optionally substituted with one or two R8a.

6. The compound of claim 5, wherein R1is monocyclic C3-8cycloalkyl optionally substituted with one or two R8a.

7. The compound of claim 6, wherein R1is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, each of which is optionally substituted with one or two R8a.

8. The compound of claim 7, wherein R1is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, each of which is optionally substituted with one R8a.

9. The compound of claim 8, wherein R1is selected from cyclobutyl and cyclohexyl, each of which is substituted with one R8a.

10. The compound of claim 1, wherein R1is C3-10heterocycloalkyl optionally substituted with one or two R8a.

11. The compound of claim 10, wherein R1is selected from aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, diazetidinyl, dioxetanyl, dithietanyl, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, isoxthiolidinyl, thiazolidinyl, isothiazolidinyl, dioxolanyl, dithiolanyl, piperidinyl, tetrahydropyranyl, diazinanyl (e.g., piperazinyl), morpholinyl, thiomorpholinyl, dioxanyl, dithianyl, azepanyl oxepanyl and thiepanyl, each of which is optionally substituted with one or two R8a.

12. The compound of claim 11, wherein R1is selected from thietanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, aziridinyl, azetidinyl, pyrrolidinyl, 86 8713541morpholinyl, piperazinyl and piperidinyl, each of which is optionally substituted with one R8a.

13. The compound of claim 12, wherein R1is selected from oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, morpholinyl, piperazinyl, and piperidinyl, each of which is optionally substituted with one R8a.

14. The compound of claim 13, R1is selected from oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is optionally substituted with one R8a.

15. The compound of claim 14, wherein R1is selected from oxetanyl, tetrahydrofuranyl, and tetrahydropyranyl, each of which is unsubstituted.

16. The compound of any one of claims 1 to 15, wherein X1is selected from CR9and N when ---- is a double bond and X1is CR9R9awhen ---- is a single bond.

17. The compound of any one of claims 1 to 16, wherein R2is H.

18. The compound of any one of claims 1 to 17, wherein X2is selected from N and CH.

19. The compound of any one of claims 1 to 18, wherein R3, R4and R5are independently selected from H, Cl, F, Br, CN, C1-4alkyl and C1-4fluoroalkyl.

20. The compound of claim 19, wherein R3, R4and R5are independently selected from H, Cl, F, CN, CH3 and CF3.

21. The compound of any one of claims 1 to 18, wherein at least one of R3and R5is selected from halo and CN.

22. The compound of any one of claims 1 to 21, wherein X3is selected from N and CH.

23. The compound of any one of claims 1 to 22, wherein R6and R7are independently selected from H, Cl, F, Br, CN, C1-4alkyl, C1-4fluoroalkyl, OC1-4alkyl and OC1-4fluoroalkyl.

24. The compound of claim 23, wherein R6and R7are independently selected from H, Cl, F, CN, CH3, CHF2,CF3, CH2CH3,CH2CH2F,OCH3, OCHF2and OCF3.

25. The compound of claim 24, wherein R6is selected from OCH3and OCF3and R7is selected from Cl, F, CH3, and CF3. 87 871354126. The compound of claim 25, wherein R6is selected from OCH3and OCF3and R7is Cl.

27. The compound of any one of claims 1 to 26, wherein R8is selected from H and CH3.

28. The compound of any one of claims 1 to 27, wherein each R8ais independently selected from OR12, NR12R13, C(O)NR12R13, Cl, F, Br, C1-4alkyl, C1-4fluoroalkyl, C3-6cycloalkyl and C3-6heterocycloalkyl, wherein all alkyl, cycloalkyl and heterocycloalkyl are optionally substituted with one or more substituents selected from Cl, Br, F, OR14, NR14R15and C1-4alkyl.

29. The compound of claim 28, wherein each R8ais independently selected from OR12, NR12R13, Cl, F, CH3, CHF2, CH3CH3 and CF3.

30. The compound of claim 29, wherein each R8ais independently selected CH3, CHF2, CH3CH3and CF3.

31. The compound of claim 29, wherein each R8ais independently selected from NR12R13and C(O)NR12R13.

32. The compound of any one of claims 1 to 16, wherein R9, R9a, R10and R11are independently selected from H, Cl, Br, F, C1-4alkyl and C1-4fluoroalkyl and R9bis selected from H, C1-4alkyl and C1-4fluoroalkyl.

33. The compound of claim 32, wherein R9, R9a, R10and R11are H and R9bis selected from H and CH3.

34. The compound of any one of claims 1 to 33, wherein R12is selected from H, C1-4alkyl, C1-4fluoroalkyl, C3-10cycloalkyl and C3-10heterocycloalkyl, the latter four groups being optionally substituted with one or two substituents selected from Cl, F, Br, OH, OCH3and OCF3.

35. The compound of claim 34, wherein R12is selected from H, CH3and CF3.

36. The compound of claim 35, wherein R12is selected from H and CH3.

37. The compound of any one of claims 1 to 36, wherein R13is selected from H, C1-4alkyl and C1-4fluoroalkyl. 88 871354138. The compound of claim 37, wherein R13is selected from H and CH3.

39. The compound of any one of claims 1 to 38, wherein R14and R15are independently selected from H, CH3and CF3.

40. The compound of claim 39, wherein R14and R15are independently selected from H and CH3.

41. The compound of claim 1, selected from: 5-Chloro-N-(4-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide; trans-5-Chloro-N-(3-(2-(((1R,4R)-4-(dimethylamino)cyclohexyl)amino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-2-methoxypyridine-3- sulfonamide formic acid salt; N-(3-(2-Amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4-fluorophenyl)-5- chloro-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin- 6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2,4-difluoro-3-(2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin- 6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2,4- difluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2,4-difluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(2-(oxetanylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)imidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(ethylamino)-8,9-dihydroimidazo[1',2':1,6]pyrido[2,3-d]pyrimidin-6-yl)-4- fluorophenyl)-2-methoxypyridine-3- 89 87135415-Chloro-N-(2-fluoro-3-(2-(methylthio)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6- yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(4-fluoro-3-(9-methyl-2-(methylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(9-methyl-2-(methylamino)-[1,2,4]triazolo[4’,3’:1,6]pyrido[2,3- d]pyrimidin-6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; trans trans-4-((6-(5-((5-Chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1- carboxamide; trans-4-((6-(3-((5-Chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)-N-methylcyclohexane-1- carboxamide; trans-4-((6-(3-((5-Chloro-2-methoxypyridine)-3-sulfonamido)-2-fluorophenyl)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-2-yl)amino)cyclohexane-1-carboxamide; ((5-Chloro-2-methoxypyridin-3-yl)sulfonyl)(4-fluoro-3-(2-(methylamino)- [1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)phenyl)amide, Potassium; 5-Chloro-N-(3-(2-(ethylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2- fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2- fluorophenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(2-fluoro-3-(2-(isopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin- 6-yl)phenyl)-2-methoxypyridine-3-sulfonamide; 5-Chloro-N-(3-(2-(cyclopropylamino)-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)- 2,4-difluorophenyl)-2-methoxypyridine-3-sulfonamide ; and N-(3-(2-Amino-[1,2,4]triazolo[4',3':1,6]pyrido[2,3-d]pyrimidin-6-yl)-2,4-difluorophenyl)-5- chloro-2-methoxypyridine-3-sulfonamide, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof.

42. A pharmaceutical composition comprising one or more compounds of any one of claims 1 to 41, or a pharmaceutically acceptable salt, solvate and / or prodrug thereof, and a pharmaceutically acceptable carrier.

43. A method of inhibiting general control nonderepressible 2 (GCN2) in a cell, either in a biological sample or in a patient, comprising administering an effective amount of one or 90 8713541more compounds of any one of claims 1 to 41, or a pharmaceutically acceptable salt, prodrug and / or solvate thereof, to the cell.

44. A method of treating a disease, disorder or condition that is treatable by inhibiting GCN2, comprising administering a therapeutically effective amount of one or more compounds of any one of claims 1 to 41, or pharmaceutically acceptable salt, prodrug and / or solvate thereof, to a subject in need thereof.

45. The method of claim 44, wherein the disease, disorder or condition that is treatable by inhibiting GCN2 is a neoplastic disorder.

46. The method of claim 44, wherein disease, disorder or condition that is treatable by inhibiting GCN2, is cancer.

47. The method of claim 46, wherein 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.

48. The method of claim 44, wherein the disease, disorder or condition that is treatable by inhibiting GCN2 is a peripheral neuropathy.

49. The method of claim 48, wherein the peripheral neuropathy is Charcot-Marie-Tooth (CMT) peripheral neuropathy.

50. A method of treating a disease, disorder or condition that is treatable by inhibiting GCN2 comprising administering a therapeutically effective amount of one or more compounds of any one of claims 1 to 41, or a pharmaceutically acceptable salt, prodrug and / or solvate thereof, in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibiting GCN2 to a subject in need thereof.

51. The method of claim 50, wherein the disease, disorder or condition treatable by inhibiting GCN2 is cancer and / or peripheral neuropathy.

52. The method of claim 50, wherein the disease, disorder or condition that is treatable by inhibiting GCN2, is cancer and the one or more compounds of the application are administered or used in combination with one or more additional cancer treatments. 91 871354153. The method of claim 52, wherein the one or more additional cancer treatments is a chemotherapeutic agent and the chemotherapeutic agent is cisplatin.

54. The method of claim 52, wherein the one or more additional cancer treatments is a chemotherapeutic agent and the chemotherapeutic agent is L-asparaginase (L-ASNase).

55. The method of claim 52, wherein the one or more additional cancer treatments is small molecule therapy and the small molecule therapy is a glutaminase inhibitor or an asparagine synthetase (ASNS) inhibitor.

56. A method of improving the efficacy of one or more cancer treatments for treating cancer comprising administering an effective amount of one or more compounds of any one of claims 1 to 41, or a pharmaceutically acceptable salt, prodrug and / or solvate thereof, in combination with an effective amount of the one or more cancer treatments.

57. The method of claim 56, wherein the one or more cancer treatments is a chemotherapeutic agent and the chemotherapeutic agent is cisplatin.

58. The method of claim 56, wherein the one or more cancer treatments is a chemotherapeutic agent and the chemotherapeutic agent is L-asparaginase (L-ASNase).

59. The method of claim 56, wherein the one or more cancer treatments is small molecule therapy and the small molecule therapy is a glutaminase inhibitor or an asparagine synthetase (ASNS) inhibitor.

60. The method of claim 56, wherein the cancer is associated with low asparagine synthetase (ASNS) expression and the one or more additional cancer treatments is L- asparaginase (L-ASNase).

61. The method of claim 56, wherein the cancer is associated with asparagine synthetase (ASNS) overexpression or dysregulation and the one or more additional cancer treatments are one or more asparagine synthetase (ASNS) inhibitors and / or L- asparaginase.

62. The method of claim 56, wherein the cancer is associated with low asparagine synthetase (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. 92 871354163. The method of claim 56, wherein cancer is associated with asparagine synthetase (ASNS) overexpression or dysregulation and glutaminase overexpression or dysregulation and the one or more additional cancer treatments are L-asparaginase (L-ASNase), one or more glutaminase inhibitors and / or one or more asparagine synthetase (ASNS) inhibitors. 93 8713541

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