Heterocyclic compounds as GCN2 and PERK kinase inhibitors

Abstract

Described herein are compounds of formula 1-A that are inhibitors of GCN2 kinase or PERK kinase, and methods of treating disorders, including GCN2 kinase or PERK kinase associated disorders, with said compounds. TIFF2024544632000067.tif6596

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 285,833, filed December 3, 2021, and U.S. Provisional Patent Application No. 63 / 348,557, filed June 3, 2022, the contents of each of which are incorporated by reference in their entirety herein. [Background technology]

[0002] Cancer cells require a continuous supply of nutrients to sustain their abnormal growth and rapid division. As part of these nutrients, amino acids are essential to support the high metabolic demands of tumor cells.

[0003] GCN2 is a serine / threonine protein kinase and one of the eukaryotic translation initiation factors 2α (eIF2α), which are key regulators of the integrated stress response (ISR). The ISR is essential for maintaining cellular homeostasis under a wide range of stressors and is activated when cells adapt to stress conditions such as hypoxia and amino acid starvation. The ISR is regulated by phosphorylation and activation of eIF2α kinases, including GCN2, which acts as an early response to perturbations in cellular homeostasis. In addition to GCN2, there are three other eIF2α kinase families, including PKR-like ER kinase (PERK), double-stranded RNA-dependent protein kinase (PKR), and heme-regulated eIF2α kinase (HRI). All four eIF2α kinases share extensive homology in their kinase catalytic domains but have distinct regulatory domains. Each of the IF2α kinases responds to distinct environmental and physiological stresses, which reflects their unique regulatory mechanisms. PERK kinase is activated under stress conditions such as ATP depletion and the unfolded protein response, and similar to GCN2, PERK kinase activation leads to upregulation of the key ISR transcription factor ATF4.

[0004] Under conditions of essential amino acid limitation or other stress factors (UV irradiation, redox stress, or proteasome inhibition), GCN2 phosphorylates eIF2α, which inhibits the formation of new ternary complexes and thus inhibits mRNA translation initiation. While decreasing overall mRNA translation, phosphorylation of eIF2α in tumor cells also increases the translation of the ISR transcription factor ATF4, which increases the expression of many stress response genes, including genes dedicated to the supply of amino acids to tumor cells, i.e., amino acid synthetases and transporters that mediate the influx of amino acids into tumor cells. ATF4 is overexpressed in human solid and liquid tumors, suggesting an important function in tumor progression.

[0005] Asparagine is a key amino acid involved in several biosynthetic pathways that significantly impact carcinogenesis and tumor biology. All cells require asparagine for their protein synthesis and growth. Normal cells obtain most of their asparagine requirements through endogenous synthesis. Compared to normal cells, cancer cells require increased amounts of asparagine to grow and proliferate, and since they cannot produce the required amount themselves, they must rely on circulating asparagine to survive. Asparagine synthetase (ASNS) catalyzes the synthesis of asparagine from aspartate and glutamine. L-asparaginase (ASNase) removes circulating asparagine, thereby depriving cancer cells of a vital nutrient and causing them to die. The use of L-asparaginase, the first example of an anticancer therapy targeting a tumor-specific metabolic signature, is an established treatment in pediatric acute lymphoblastic leukemia (ALL), but toxicity limits its use beyond this patient population. The particularly low level of expression of ASNS in many ALL cell lines makes asparagine depletion an effective therapeutic method due to the cells' abnormal dependence on circulating serum asparagine as a nutrient required for growth compared to that of normal cells. A poor response to asparaginase is associated with an increased risk of relapse. Other hematological and solid cancers express low levels of ASNS and therefore should also be asparagine auxotrophs and asparaginase sensitive. Conversely, in some cancer types, ASNS is overexpressed, promoting cell proliferation, chemoresistance, and metastatic behavior. In the case of asparaginase-resistant cancers, the action of blood asparagine depletion through L-asparaginase instead leads to significant ASNS overexpression, virtually negating the effects of chemotherapeutic agents. Numerous studies have shown that ASNS is at the heart of the cellular response to amino acid deprivation and other forms of cellular stress. Through transcriptional regulation, the ASNS gene is the target of two signaling pathways aimed at ensuring cell survival. The first, termed the Amino Acid Response (AAR), is activated by GCN2 kinase under conditions of imbalanced amino acid availability.The second pathway, termed the Unfolded Protein Response (UPR), is activated by PERK kinase under conditions of increased endoplasmic reticulum stress. The AAR and UPR pathways converge on the phosphorylation of eIF2α, which leads to preferential translation of a select population of mRNAs, including the transcription factor ATF4, with concomitant attenuation of global protein synthesis. ATF4 is the primary driver of ASNS induction and functions as a transactivator through binding to enhancer elements within the ASNS promoter.

[0006] GCN2 sensitizes cancer cells with low basal expression of ASNS to the anti-leukemic agent L-asparaginase in vitro and in vivo. Treatment with GCN2 inhibitors sensitized acute lymphoblastic leukemia cells to L-asparaginase by preventing the induction of ASNS. GCN2 inhibitors exhibit synergistic antiproliferative effects with L-asparaginase in ASNS-low / deficient cancers. Thus, combination therapy of GCN2 inhibitors and L-asparaginase is promising for achieving improved outcomes in acute lymphoblastic leukemia and other types of cancer. Acute lymphoblastic leukemia, acute myeloid leukemia, and pancreatic cancer cells are particularly sensitive to combination therapy of L-asparaginase and GCN2 inhibitors. Previously reported studies have shown that combination therapy of ASNase and GCN2 inhibitors in acute lymphoblastic leukemia, acute myeloid leukemia, and pancreatic cancer cells showed potent antitumor activity compared with the results of monotherapy with L-asparaginase or GCN2 inhibitors. Thus, GCN2 inhibitors may represent sensitizers to L-asparaginase used in the treatment of these tumors. In summary, GCN2 inhibition enhances the sensitivity of cancer cells with low basal ASNS expression to L-asparaginase treatment by preventing ASNS induction.

[0007] Inhibition of GCN2 may also be an effective strategy for targeting the tumor microenvironment, including the immune system, including tryptophan-dependent immune surveillance of tumor cells.

[0008] The tumor microenvironment (TME: a set of extracellular components and stromal cells (endothelial cells, cancer-associated fibroblasts, tumor-associated macrophages, tumor-infiltrating T cells) that surrounds tumor cells is characterized by a lack of oxygen and key nutrients such as glucose and amino acids, resulting in an overall immunosuppressive environment.

[0009] Many tumors have evolved to evade immune surveillance by exploiting their metabolic flexibility and redirecting nutrients to their own benefit. Stromal cells and myeloid-derived suppressor cells (MDSCs) within the tumor inhibit immune function and create a nutrient-deprived environment that supports tumor growth.

[0010] Increased catabolism of tryptophan, an essential amino acid, driven by overexpression of key enzymes in tryptophan metabolism [indoleamine-2,3-dioxygenase (IDO) and tryptophan-2,3-dioxygenase (TDO)] by cells in the tumor microenvironment leads to an immunosuppressive microenvironment in many types of cancer. Local tryptophan depletion is thought to be a crucial T cell immunosuppressive mechanism. In T cells, GCN2 kinase has been identified as a molecular sensor of tryptophan depletion. Activation of GCN2 by tryptophan depletion induces apoptosis and attenuates T cell proliferation. GCN2 is a key effector signaling component of IDO / TDO and is considered a highly tryptophan-dependent T cell metabolic checkpoint.

[0011] The GCN2 pathway is not only important for tumor immune evasion, but also plays an active role in regulating other aspects of the tumor microenvironment. It has been demonstrated that GCN2 knockdown prevents amino acid deprivation (AAD)-induced expression of vascular endothelial growth factor (VEGF), which tumors use to enhance nutrient supply via increased angiogenesis. Thus, activation of the GCN2 / ATF4 pathway promotes tumor growth and angiogenesis through AAD-mediated VEGF expression. Abrogation of ATF4 or GCN2 expression significantly inhibited tumor growth in vivo.

[0012] Therefore, selective inhibition of GCN2 can increase the activity of the immune system and reduce the angiogenesis of the tumor microenvironment. The GCN2-eIF2α-ATF4 pathway is important for maintaining the metabolic homeostasis of tumor cells under stress conditions and for maintaining the microenvironment of immunosuppressive immune cells. The PERK-ATF4 pathway is also important for maintaining the homeostasis of tumor cells under stress conditions. It has been reported that there is crosstalk regulation of both GCN2 and PERK signaling pathways, such that inhibition of GCN2 activates PERK as a compensatory mechanism, and vice versa.

[0013] There is a need for inhibitors of GCN2 and / or PERK that modulate the tumor-promoting aspects of GCN2 and / or PERK both in the tumor cell (tumor cell autonomous) and in the tumor immune cell microenvironment. Summary of the Invention

[0014] Described herein are compounds that modulate (inhibit or activate) GCN2 (general control nonderepressible 2) kinase and / or PERK (PKR-like ER kinase) kinase, and methods of their use for the treatment of disorders, including GCN2- or PERK-related diseases.

[0015] In one embodiment, the compound represented by formula IA: [ka] or a pharma- ceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, as described herein, wherein: X 1 and X 3 are each independently selected from the group consisting of CH and N; 2 is NR 6 , O, and S; R 1 , R 2 , and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4is selected from the group consisting of halogen, alkoxy and alkyl; R 5 is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acyl.

[0016] In another embodiment, described herein are pharmaceutical compositions comprising a compound described herein (e.g., compounds of formulas IA, IB, IC, and ID described herein) and a pharma- ceutically acceptable carrier or excipient.

[0017] In another embodiment, described herein is a method of treating a disease caused by dysregulation of the integrated stress response in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0018] In another embodiment, described herein are methods of treating a disease caused by dysregulation of the integrated stress response and / or the unfolded protein response in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0019] In another embodiment, described herein is a method of modulating the activity of GCN2 kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0020] In another embodiment, described herein are methods of activating GCN2 kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0021] In another embodiment, described herein is a method of modulating activity of PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0022] In another embodiment, described herein are methods of activating PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0023] In another embodiment, described herein is a method of inhibiting GCN2 kinase and PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0024] In another embodiment, described herein are methods of modulating the activity of GCN2 kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0025] In another embodiment, described herein are methods of inhibiting the activity of PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0026] In another embodiment, described herein is a method of treating a disease selected from a GCN2-related disease and a PERK-related disease in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0027] In another embodiment, described herein is a method of treating a disease selected from a GCN2-related disease and a PERK-related disease in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, and a therapeutically effective amount of one or more therapeutic agents.

[0028] In another embodiment, described herein are methods of treating cancer in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., compounds of formulas IA, IB, IC, and ID described herein), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0029] In another embodiment, described herein is a method of treating a disorder selected from the group consisting of melanoma, fibrosarcoma, thyroid cancer, ovarian cancer, colon cancer, pancreatic cancer, lung cancer, bladder cancer, gastrointestinal stromal tumors, solid tumors, blood-borne cancers, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), and other cancers caused by activation of the GCN2 signaling pathway, in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., compounds of formulas IA, IB, IC, and ID described herein) or a pharma- ceutically acceptable salt thereof. [Brief description of the drawings]

[0030] [Figure 1] 1 is a graphical representation showing the unexpected stimulation of the UPR / ISR marker ATF4 (black bars) in response to increasing concentrations of Compound 2. [Diagram 2]1 shows a graph of stimulation of PERK oligomers induced by Compound 2 in a NanoBRET assay. [Diagram 3] Shows stimulation of PERK downstream signaling proteins ATF4 and CHOP (actin as a loading control) in H929 multiple myeloma cells. [Figure 4] 1 is a graphical representation showing stimulation of ATF4 target genes in response to increasing concentrations of Compound 2 in H929 multiple myeloma cells. [Diagram 5] FIG. 1 shows the effect of Compound 2 on readout of apoptotic pathways in H929 multiple myeloma cells. [Figure 6A] Figures 6A, 6B, 6C and 6D show cell proliferation studies of compound 2 in combination with standard of care in RPMI8226 multiple myeloma cells (Figure 6A), H929 multiple myeloma cells (Figure 6B), GA-10 Burkitt's lymphoma cells (Figure 6C), and follicular lymphoma cells (Figure 6D). [Figure 6B] Figures 6A, 6B, 6C and 6D show cell proliferation studies of compound 2 in combination with standard of care in RPMI8226 multiple myeloma cells (Figure 6A), H929 multiple myeloma cells (Figure 6B), GA-10 Burkitt's lymphoma cells (Figure 6C), and follicular lymphoma cells (Figure 6D). [Figure 6C] Figures 6A, 6B, 6C and 6D show cell proliferation studies of compound 2 in combination with standard of care in RPMI8226 multiple myeloma cells (Figure 6A), H929 multiple myeloma cells (Figure 6B), GA-10 Burkitt's lymphoma cells (Figure 6C), and follicular lymphoma cells (Figure 6D). [Figure 6D] Figures 6A, 6B, 6C and 6D show cell proliferation studies of compound 2 in combination with standard of care in RPMI8226 multiple myeloma cells (Figure 6A), H929 multiple myeloma cells (Figure 6B), GA-10 Burkitt's lymphoma cells (Figure 6C), and follicular lymphoma cells (Figure 6D). [Figure 7A]Figures 7A and 7B show the effect of Compound 2 in combination with asparaginase on ATF4 levels, which measures GCN2 activity, in an in vivo PK / PD model. Figure 7A shows a data plot of the results from the PK / PD model study, and Figure 7B shows the results in tabular format. [Figure 7B] Figures 7A and 7B show the effect of Compound 2 in combination with asparaginase on ATF4 levels, which measures GCN2 activity, in an in vivo PK / PD model. Figure 7A shows a data plot of the results from the PK / PD model study, and Figure 7B shows the results in tabular format. [Figure 8] 1 shows plots demonstrating the effect of Compound 2 in combination with asparaginase on tumor growth in an MV-4-11 xenograft model. [Figure 9A] Figure 9A shows the effect of Compound 2 in a recombinant GCN2 activity assay, which measures the activity of recombinant GCN2 enzyme in vitro. Figure 9B shows the effect of Compound 2 in a recombinant PERK activity assay, which measures the activity of recombinant PERK enzyme in vitro. [Figure 9B] Figure 9A shows the effect of Compound 2 in a recombinant GCN2 activity assay, which measures the activity of recombinant GCN2 enzyme in vitro. Figure 9B shows the effect of Compound 2 in a recombinant PERK activity assay, which measures the activity of recombinant PERK enzyme in vitro. [Figure 10A] Figures 10A, 10B, and 10C show the effect of Compound 2 on the ISR and apoptotic pathway in solid tumor cancer (Figure 10A, H2030; Figure 10B, H358; Figure 10C, HT-1080) cell lines. [Figure 10B] Figures 10A, 10B, and 10C show the effect of Compound 2 on the ISR and apoptotic pathway in solid tumor cancer (Figure 10A, H2030; Figure 10B, H358; Figure 10C, HT-1080) cell lines. [Figure 10C] Figures 10A, 10B, and 10C show the effect of Compound 2 on the ISR and apoptotic pathway in solid tumor cancer (Figure 10A, H2030; Figure 10B, H358; Figure 10C, HT-1080) cell lines. [Figure 11A] Figures 11A, 11B, and 11C show the effect of Compound 2 in a spheroid proliferation assay. Figure 11A shows cell regression in H2030 solid tumor spheroids by Compound 2 as a single agent. Figures 11B and 11C show the effect of Compound 2 as a single agent on cell proliferation in H358 (Figure 11B) and HT-1080 (Figure 11C) solid tumor spheroids. [Figure 11B] Figures 11A, 11B, and 11C show the effect of Compound 2 in a spheroid proliferation assay. Figure 11A shows cell regression in H2030 solid tumor spheroids by Compound 2 as a single agent. Figures 11B and 11C show the effect of Compound 2 as a single agent on cell proliferation in H358 (Figure 11B) and HT-1080 (Figure 11C) solid tumor spheroids. [Figure 11C] Figures 11A, 11B, and 11C show the effect of Compound 2 in a spheroid proliferation assay. Figure 11A shows cell regression in H2030 solid tumor spheroids by Compound 2 as a single agent. Figures 11B and 11C show the effect of Compound 2 as a single agent on cell proliferation in H358 (Figure 11B) and HT-1080 (Figure 11C) solid tumor spheroids. [Figure 12A] Figures 12A, 12B, and 12C show the effect of compound 2 in combination with standard therapeutic agents for the inhibition of spheroid growth. Figure 12A shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without sotorasib. Figure 12B shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without sotorasib. Figure 12C shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without trametinib. [Figure 12B]Figures 12A, 12B, and 12C show the effect of compound 2 in combination with standard therapeutic agents for the inhibition of spheroid growth. Figure 12A shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without sotorasib. Figure 12B shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without sotorasib. Figure 12C shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without trametinib. [Figure 12C] Figures 12A, 12B, and 12C show the effect of compound 2 in combination with standard therapeutic agents for the inhibition of spheroid growth. Figure 12A shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without sotorasib. Figure 12B shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without sotorasib. Figure 12C shows cell proliferation experiments of H2030 NSCLC spheroids treated with compound 2 titers in combination with or without trametinib. [Figure 13A] Figures 13A, 13B, and 13C show the effect of Compound 2 in HT-1080 shRNA knockdown assay. Figure 13A shows GCN2 and PERK knockdown using targeted shRNA in HT-1080 cell line. Figure 13B shows Western blot quantification of ATF4 signal. Figure 13C shows the effect of Compound 2 on spheroid growth inhibition. [Figure 13B] Figures 13A, 13B, and 13C show the effect of Compound 2 in HT-1080 shRNA knockdown assay. Figure 13A shows GCN2 and PERK knockdown using targeted shRNA in HT-1080 cell line. Figure 13B shows Western blot quantification of ATF4 signal. Figure 13C shows the effect of Compound 2 on spheroid growth inhibition. [Figure 13C]Figures 13A, 13B, and 13C show the effect of Compound 2 in HT-1080 shRNA knockdown assay. Figure 13A shows GCN2 and PERK knockdown using targeted shRNA in HT-1080 cell line. Figure 13B shows Western blot quantification of ATF4 signal. Figure 13C shows the effect of Compound 2 on spheroid growth inhibition. [Figure 14A] Figures 14A and 14B show the effect of Compound 2 in the HT-1080 fibrosarcoma xenograft PK / PD model. Figure 14A shows the upregulation of ATF4 levels by Compound 2 at different doses. Figure 14B shows the corresponding plasma levels of Compound 2 at various time points after administration. [Figure 14B] Figures 14A and 14B show the effect of Compound 2 in the HT-1080 fibrosarcoma xenograft PK / PD model. Figure 14A shows the upregulation of ATF4 levels by Compound 2 at different doses. Figure 14B shows the corresponding plasma levels of Compound 2 at various time points after administration. [Figure 15A] Figures 15A and 15B show the effect of Compound 2 in the HT-1080 xenograft efficacy model. Figure 15A shows the effect of Compound 2 on tumor growth in the LoVo colorectal xenograft model, and Figure 15B shows the effect of Compound 2 on tumor growth in the HT-1080 fibrosarcoma xenograft model. [Figure 15B] Figures 15A and 15B show the effect of Compound 2 in the HT-1080 xenograft efficacy model. Figure 15A shows the effect of Compound 2 on tumor growth in the LoVo colorectal xenograft model, and Figure 15B shows the effect of Compound 2 on tumor growth in the HT-1080 fibrosarcoma xenograft model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0031] The features and other details of the present disclosure are described in more detail below.Specific terms employed in the specification, examples and appended claims are summarized here.These definitions should be read in light of the remainder of this disclosure as understood by those skilled in the art.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0032] definition The definitions set forth in this application are intended to clarify terms used throughout this application.

[0033] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this subject matter belongs. As used in this specification and the appended claims, unless otherwise specified, the following terms have the meanings set forth to facilitate understanding of this disclosure.

[0034] When a bond to a substituent is shown to cross a bond connecting two atoms in a ring, such a substituent may be bonded to any atom on the ring. When a substituent is listed without indicating the atom through which such a substituent is bonded to the remainder of the compound of a given formula, such a substituent may be bonded through any atom in such a substituent. Combinations of substituents, positions of substituents and / or variables are permissible if such combinations result in stable compounds.

[0035] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0036] As used herein, the term "herein" means the entire application.

[0037] As used herein, "deuterated" means that at least one hydrogen atom is replaced by deuterium. In any sample of a deuterated compound, it is likely that some individual molecules of the compound will have hydrogen rather than deuterium at the specified position. However, the percentage of molecules of the deuterated compound that have deuterium at the specified position will be much greater than that which occurs naturally. Deuterium at the deuterated position is enriched.

[0038] As used herein, the term "optionally" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes cases where the event or circumstance occurs as well as cases where it does not occur. For example, "optionally substituted alkyl" refers to cases where the alkyl may be substituted and also to cases where the alkyl is not substituted.

[0039] It will be appreciated that the substituents and substitution patterns on the compounds of the present disclosure can be selected by one of ordinary skill in the art to provide chemically stable compounds that can be readily synthesized from readily available starting materials by techniques known in the art, as well as the methods described below. When a substituent is itself substituted with multiple groups, it is understood that these multiple groups can be on the same carbon or different carbons, so long as a stable structure is obtained.

[0040] As used herein, the term "optionally substituted" refers to the replacement of 1-6 hydrogen atoms in a given structure with the radical of a specified substituent, including, but not limited to, hydroxy, hydroxyalkyl, alkoxy, halogen, alkyl, aryl, cycloalkyl, heterocyclyl, amino, aminoalkyl, cyano, haloalkyl, haloalkoxy, -OC(=O)-CH2-Oalkyl. Preferably, "optionally substituted" refers to the replacement of 1-4 hydrogen atoms in a given structure with a substituent as described above. More preferably, 1-3 hydrogen atoms are replaced by a substituent as described above. It is understood that the substituents may be further substituted.

[0041] As used herein, the term "substituted" refers to a moiety having a substituent replacing a hydrogen on one or more carbons of the backbone. It should be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is in accordance with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound that is not naturally transformed, e.g., by rearrangement, cyclization, elimination, and the like. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more of the same or different substituents for appropriate organic compounds. For purposes of this application, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituent of organic compounds described herein that satisfies the valences of the heteroatoms.

[0042] Substituents can include any of the substituents described herein, for example, such substituents can include, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), alkoxy, amino, amido, imine, cyano, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties, unless otherwise specified. Those skilled in the art will appreciate that the substituents themselves can be substituted, where appropriate. For example, the substituents of substituted alkyl can include amino, amido, sulfonyl, as well as substituted and unsubstituted forms of ethers, carbonyls (including carboxylates and esters), -CF3, -CN, and the like. References to chemical moieties herein are understood to include substituted variants, unless specifically described as "unsubstituted." For example, references to an "aryl" group or moiety implicitly include both substituted and unsubstituted variants.

[0043] As used herein, the term "alkyl" refers to a fully saturated straight or branched chain non-aromatic hydrocarbon. Typically, a straight or branched chain alkyl group has from 1 to about 20, preferably from 1 to about 10, carbon atoms, e.g., C1-C6, unless otherwise defined. 10 It may be an alkyl or, for example, a C1-C6 alkyl. Examples of straight chain and branched alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl (n-propyl), 2-propyl, n-butyl, sec-butyl, tert-butyl, 1-pentyl, 2-pentyl, 3-pentyl, neo-pentyl, 1-hexyl, 2-hexyl, 3-hexyl, 1-heptyl, 2-heptyl, 3-heptyl, 4-heptyl, 1-octyl, 2-octyl, 3-octyl, or 4-octyl. Furthermore, the term "alkyl" as used throughout the specification, examples, and claims is intended to include both "unsubstituted alkyl" and "substituted alkyl," the latter referring to an alkyl moiety having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. An "alkyl" group may be optionally substituted.

[0044] When used with chemical moieties such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, "C x -C y " is meant to include groups containing x to y carbons in the chain. For example, "C x -C y The term " refers to a substituted or unsubstituted saturated hydrocarbon group, including straight and branched chain alkyl groups containing x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl. C0 alkyl indicates a hydrogen when the group is in a terminal position and a bond when it is internal.

[0045] As used herein, the term "alkenyl" refers to an unsaturated straight or branched chain hydrocarbon having at least one carbon-carbon double bond. Exemplary alkenyl groups include, but are not limited to, straight or branched chain groups of 2 to 6 or 3 to 4 carbon atoms, referred to herein as C2-C6 alkenyl and C3-C4 alkenyl, respectively. Exemplary alkenyl groups include, but are not limited to, vinyl, allyl, butenyl, pentenyl, and the like.

[0046] As used herein, the term "alkynyl" refers to an unsaturated straight or branched chain hydrocarbon having at least one carbon-carbon triple bond. Exemplary alkynyl groups include, but are not limited to, straight or branched groups of 2 to 6 or 3 to 6 carbon atoms, referred to herein as C2-C6 alkynyl and C3-C6 alkynyl, respectively. Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, methylpropynyl, and the like.

[0047] As used herein, the term "alkoxy" refers to a straight or branched chain saturated aliphatic (alkyl) hydrocarbon radical bonded to an oxygen atom attached to a core structure. The alkoxy group preferably has 1-6 carbon atoms, i.e., may be a C1-C6 alkoxy. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentoxy, 3-methylbutoxy, and the like.

[0048] As used herein, the term "alkoxyalkyl" refers to an alkyl group (as defined above) substituted with an alkoxy group, and may be represented by the general formula alkyl-O-alkyl. Examples of alkoxyalkyl groups include, but are not limited to, methyl-O-ethylene-, ethyl-O-ethylene-.

[0049] As used herein, the term "haloalkyl" refers to an alkyl group (defined above) substituted with one or more halogens. Monohaloalkyl radicals can have, for example, a chlorine, bromine, iodine, or fluorine atom. Dihalo and polyhaloalkyl radicals can have two or more of the same or different halogen atoms. Examples of haloalkyl include, but are not limited to, chloromethyl, dichloromethyl, trichloromethyl, dichloroethyl, dichloropropyl, fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl, difluoroethyl, difluoropropyl, and the like.

[0050] As used herein, the term "haloalkoxy" refers to a radical in which one or more of the hydrogen atoms of an alkoxy group have been replaced with one or more halogens. Representative examples of "haloalkoxy" groups include, but are not limited to, difluoromethoxy (-OCHF2), trifluoromethoxy (-OCF3), or trifluoroethoxy (-OCH2CF3).

[0051] As used herein, the term "aryl" includes substituted or unsubstituted monocyclic aromatic groups in which each atom of the ring is carbon. The ring is preferably 5-7 membered, more preferably 6 membered. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings (fused rings), at least one of the rings being aromatic. For example, the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. The term "fused" means that a second ring is attached to or formed by having two adjacent atoms in common with the first ring. The term "fused" is equivalent to the term "fused". Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthryl, phenol, aniline, indanyl, dihydrobenzofuranyl, dihydroisobenzofuranyl, indolinyl, isoindolinyl, and the like. Unless otherwise specified, aryl groups described herein may be optionally substituted.

[0052] As used herein, the terms "polycyclyl," "polycycle," and "polycyclic" refer to two or more rings (e.g., cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl) in which one or more atoms are common to two or more adjacent rings. For example, the rings are "fused rings." Each of the rings of a polycycle can be substituted or unsubstituted. In certain embodiments, each ring of a polycycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

[0053] As used herein, the term "acyl" refers to a -C(=O)-R w In the formula, R w is optionally substituted alkyl. Examples of "acyl" include, but are not limited to, R w But, C1-C 10 Alkyl (C1-C 10 acyl) or C1-C 6-Examples include alkyl (C1-C6 acyl). In some embodiments, each occurrence of an optionally substituted substituent is selected from the group consisting of H, OH, alkoxy, cyano, F, and amino. Additional examples of "acyl" include -C(=O)-CH3, -C(=O)-CH2-CH3, -C(=O)-CH2-CH2-CH3, or -C(=O)-CH(CH3)2.

[0054] As used herein, the terms "amine" and "amino" refer to both unsubstituted and substituted amines and their salts, e.g., moieties represented by the formula: [ka] In the formula, each R z independently represent hydrogen or a hydrocarbyl group, or R z together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure.

[0055] As used herein, the terms "amide" and "amido" are represented by the formula: [ka] In the formula, R x , R y , and R z each independently represents hydrogen or a hydrocarbyl group, or R y , and R z together with the N atom to which they are attached complete a heterocyclyl having 4 to 8 atoms in the ring structure.

[0056] As used herein, the term "acylamino" refers to an amino group, as defined above, substituted with an acyl group.

[0057] As used herein, "aminocarbonyl" refers to a carbonyl group substituted with an amino group.

[0058] As used herein, the term "alkenylalkyl" refers to an alkyl group substituted with an alkenyl group.

[0059] As used herein, the term "alkynylalkyl" refers to an alkyl group substituted with an alkynyl group.

[0060] As used herein, the term "alkylamino" refers to an amino group, as defined above, substituted with at least one alkyl group.

[0061] As used herein, the term "aminoalkyl" refers to an alkyl group substituted with an amino group.

[0062] As used herein, the term "amidoalkyl" refers to an alkyl group substituted with an amide group.

[0063] As used herein, the term "cyanoalkyl" refers to an alkyl group substituted with a cyano group.

[0064] As used herein, the term "cycloalkoxyalkyl" refers to an alkyl group (as defined above) substituted with a cycloalkoxy group, and may be represented by the general formula cycloalkyl-O-alkyl. Examples of cycloalkoxyalkyl groups include, but are not limited to, cyclopropyl-O-methylene-, cyclopropyl-O-ethylene.

[0065] As used herein, the term "cycloalkylalkyl" refers to an alkyl group substituted with a cycloalkyl group.

[0066] As used herein, the term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group.

[0067] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group.

[0068] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group.

[0069] As used herein, the term "cycloalkyl," alone or in combination with other terms, refers to a fully saturated cyclic hydrocarbon. "Cycloalkyl" includes monocyclic, bicyclic, and tricyclic rings. Typically, monocyclic cycloalkyl groups, unless otherwise defined, have from 3 to about 10 carbon atoms, more typically from 3 to 8 carbon atoms (e.g., C3-C4). 10 Cycloalkyl, or, for example, C3-C6 cycloalkyl). Examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. The second ring of a bicyclic cycloalkyl, or the second or third ring of a tricyclic cycloalkyl, can be selected from saturated, unsaturated, and aromatic rings. Cycloalkyl includes bicyclic and tricyclic molecules in which one, two, or three or more atoms are shared between the two rings. Cycloalkyl may be further substituted with alkyl, alkenyl, alkoxy, aminoalkyl, carbonyl-substituted alkyl, -CF3, -CN, and the like.

[0070] As used herein, the term "cycloalkylalkyl" refers to an alkyl group substituted with a cycloalkyl group.

[0071] As used herein, the term "cyano" refers to the group --CN.

[0072] As used herein, the term "hydroxy" or "hydroxyl" refers to an --OH group.

[0073] The terms "halo" or "halogen," as used herein, alone or in combination with other terms, mean chloro, fluoro, bromo, and iodo.

[0074] As used herein, the term "heteroatom" refers to an atom of any element other than carbon or hydrogen. Exemplary heteroatoms are nitrogen (N), oxygen (O), sulfur (S), and silicon (Si).

[0075] As used herein, the terms "heterocyclyl," "heterocycloalkyl," "heterocycle," and "heterocyclic" refer to a non-aromatic, saturated, or partially saturated ring system, including 3-15 membered, monocyclic, polycyclic (e.g., bicyclic, tricyclic), bridged or fused, having at least one heteroatom or heterogroup selected from O, N, S, S(O), S(O), NH, or C(O), with the remaining ring atoms independently selected from the group consisting of carbon, oxygen, nitrogen, and sulfur. Examples of "heterocycloalkyl" include, but are not limited to, azetidinyl, oxetanyl, imidazolidinyl, pyrrolidinyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,4-dioxanyl, dioxidothiomorpholinyl, oxapiperazinyl, oxapiperidinyl, tetrahydrofuryl, tetrahydropyranyl, tetrahydrothiophenyl, dihydropyranyl, indolinyl, indolinylmethyl, 2-azabicyclo[2.2.2]octanyl, azocinyl, chromanyl, xanthenyl, and N-oxides thereof. Attachment of the heterocycloalkyl substituent can occur via either a carbon atom or a heteroatom. Heterocycloalkyl groups may be optionally substituted with one or more suitable groups, such as one or more of the aforementioned groups. Preferably, "heterocycloalkyl" refers to a 5- or 6-membered ring selected from the group consisting of azetidinyl, oxetanyl, imidazolidinyl, pyrrolidinyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,4-dioxanyl, and N-oxides thereof. More preferably, "heterocycloalkyl" includes azetidinyl, pyrrolidinyl, morpholinyl, and piperidinyl. Heterocycloalkyl is optionally substituted with one or more of the aforementioned groups.

[0076] As used herein, the term "heteroaryl" refers to a substituted or unsubstituted aromatic single ring structure, preferably a 5-7 membered ring, more preferably a 5-6 membered ring, the ring structure containing at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The term "heteroaryl" also refers to a substituted or unsubstituted aromatic or partially aromatic ring system containing at least one heteroatom, having two or more cyclic rings (bicyclic, tricyclic, or polycyclic), containing 8-20 ring atoms, preferably 5-10 ring atoms, which are covalently bonded or fused, two or more atoms are common to two adjacent rings, and at least one of the rings is heteroaromatic, e.g., the other cyclic ring may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, and / or heterocyclyl. The ring may contain N or S atoms, the N or S atoms being optionally oxidized, or the N atoms being optionally quaternized. All heteroaryls are optionally substituted. Any suitable ring position of the heteroaryl moiety may be covalently linked to the defined chemical structure. Examples of heteroaryls include furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, cinnolinyl, isoxazolyl, thiazolyl, isothiazolyl, 1H-tetrazolyl, oxadiazolyl, thiadiazolyl, triazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl, benzofuranyl, benzothienyl, benzotriazinyl, phthalazinyl, thianthrene, dibenzofuranyl, dibenzothienoyl, ... Examples of the aryl radicals include, but are not limited to, benzimidazolyl, indolyl, isoindolyl, indazolyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, purinyl, pteridinyl, 9H-carbazolyl, α-carboline, indolizinyl, benzisothiazolyl, benzoxazolyl, pyrrolopyridyl, furopyridinyl, purinyl, benzothiadiazolyl, benzoxadiazolyl, benzotriazolyl, benzotridiazolyl, carbazolyl, dibenzothienyl, acridinyl, and the like.

[0077] As used herein, the term "hydrocarbyl" refers to a group that does not have =O or =S substituents and typically has at least one carbon-hydrogen bond and a predominantly carbon backbone, but may optionally contain heteroatoms, bonded through a carbon atom. Thus, groups such as methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered hydrocarbyl for the purposes of this application, while substituents such as acetyl (having =O substituent on the bonded carbon) and ethoxy (bonded through an oxygen rather than a carbon) are not hydrocarbyl. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, cycloalkyl, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

[0078] As used herein, the term "sulfonamide" is represented as follows: [ka] In the formula, R z represents independently at each occurrence a hydrogen, an alkyl, or a cycloalkyl group, or R z The groups, together with the N atom to which they are attached, complete a heterocycle having 4 to 8 atoms in the ring structure.

[0079] As used herein, the term "sulfonyl" refers to -S(O)-R 6d R refers to the group 6d represents alkyl or cycloalkyl.

[0080] "Combination therapy" is a treatment that includes the administration of two or more therapeutic agents, for example a compound of Formula I and the enzyme asparaginase (ASNase) or a derivative thereof, to a patient in need of treatment.

[0081] "Disease," "disorder," and "condition" are used interchangeably herein.

[0082] An "individual," "patient," or "subject" is used interchangeably herein and includes any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, most preferably a human. The compounds described herein can be administered to mammals, such as humans, but can also be administered to other mammals, such as animals requiring veterinary treatment, for example, companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.).

[0083] The compounds described herein are useful in treating GCN2-driven diseases (sometimes abbreviated herein as "GCN2-associated diseases"), such as cancer (e.g., colorectal cancer (e.g., colorectal cancer, rectal cancer, anal cancer, familial colorectal cancer, hereditary nonpolyposis colorectal cancer, gastrointestinal stromal tumors), lung cancer (e.g., non-small cell lung cancer, small cell lung cancer, malignant mesothelioma), mesothelioma, pancreatic cancer (e.g., pancreatic ductal carcinoma, pancreatic endocrine tumors) , pharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer (e.g., papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma), duodenal cancer, small intestine cancer, breast cancer (e.g., invasive ductal carcinoma, ductal carcinoma in situ, inflammatory breast cancer), ovarian cancer (e.g., ovarian epithelial cancer, extragonadal germ cell tumor, ovarian germ cell tumor, ovarian low malignant potential tumor), testicular tumor, prostate cancer (e.g., hormone-dependent prostate cancer, non-hormone-dependent prostate cancer, castration-resistant prostate ), liver cancer (e.g., hepatocellular carcinoma, primary liver cancer, extrahepatic bile duct cancer), thyroid cancer (e.g., medullary thyroid cancer), kidney cancer (e.g., renal cell carcinoma (e.g., renal clear cell carcinoma), transitional cell carcinoma of the renal pelvis and ureter), uterine cancer (e.g., cervical cancer, endometrial cancer, uterine sarcoma), gestational choriocarcinoma, brain tumors (e.g., medulloblastoma, glioma, pineal astrocytoma, pilocytic astrocytoma, diffuse astrocytoma, anaplastic astrocytoma, ptosis, The present invention is useful for the treatment of cancers including, for example, myeloma, thyroid cancer, thyroid cancer, thyroid cancers, and thyroid cancers of unknown primary nucleus, as well as for the treatment of cancers of unknown primary nucleus, such as myeloma, thyroid cancer, thyroid cancers of unknown primary nucleus, thyroid cancers of unknown cellular origin ...

[0084] The compounds described herein, e.g., compounds of formula I as defined herein, may be used in combination with one or more additional therapeutic agents to treat the disorders described herein, such as the cancers described herein. In some embodiments, the compounds described herein may be used in combination with hormonal therapeutic agents, chemotherapeutic agents, immunotherapeutic agents, agents that inhibit the action of cell growth factors and their receptors, such as PERK inhibitors and autophagy inhibitors, the enzyme asparaginase (ASNase), and the like.

[0085] "Pharmaceutically or pharmacologically acceptable" includes molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to animals or humans, as appropriate. For human administration, preparations should meet sterility, pyrogenicity, and general safety and purity standards as required by FDA Office of Biologics standards.

[0086] As used herein, the term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers to any and all solvents, dispersion media, coatings, isotonicity agents, absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The composition may also include other active compounds that provide supplementary, additional, or enhanced therapeutic functions.

[0087] As used herein, the term "pharmaceutical composition" refers to a composition comprising at least one compound disclosed herein formulated together with one or more pharma- ceutically acceptable carriers.

[0088] As used herein, the term "pharmaceutically acceptable salts" refers to salts of acidic or basic groups that may be present in the compounds used in the compositions. Compounds included in the compositions that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids which may be used to prepare pharma- ceutically acceptable acid addition salts of such basic compounds are those which form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including, but not limited to, malate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Compounds contained in the present composition that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, specifically calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts. Compounds contained in the present composition that contain a basic or acidic moiety can also form pharma-ceutically acceptable salts with various amino acids. Compounds of the present disclosure may contain both acidic and basic groups, for example, one amino group and one carboxylic acid group. In such cases, the compounds may exist as acid addition salts, zwitterions, or base salts.

[0089] The compounds of the present disclosure may contain one or more chiral centers and therefore may exist as stereoisomers. As used herein, the term "stereoisomers" consists of all enantiomers or diastereomers. These compounds may be designated by the symbols "R" or "S" depending on the configuration of the substituents around the stereogenic carbon atom, although those skilled in the art will recognize that the structure may implicitly indicate a chiral center. These compounds may also be designated by "(+)" and "(-)" based on their optical rotation. The compounds described herein encompass the various stereoisomers of these compounds and mixtures thereof. Mixtures of enantiomers or diastereomers may be designated by the symbols "(±)" in nomenclature, although those skilled in the art will recognize that the structure may implicitly indicate a chiral center.

[0090] As used herein, the term "therapeutically effective amount" refers to an amount of the subject compound that elicits the biological or medical response of a tissue, system, or animal (e.g., a mammal or human) that is desired by a researcher, veterinarian, physician, or other clinician. The compounds described herein are administered in a therapeutically effective amount to treat a disorder.

[0091] "Treating" includes any effect that results in the improvement of a condition, disease, disorder, or the like, eg, amelioration, alleviation, modulation, or elimination.

[0092] The present disclosure also encompasses isotopically labeled compounds that are identical to those enumerated herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from that normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, e.g., 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35S, 18 F, and 36 Cl. For example, compounds of the present disclosure may have one or more H atoms replaced with deuterium.

[0093] Individual enantiomers and diastereomers of the disclosed compounds may be prepared synthetically from commercially available starting materials containing asymmetric or stereocenters, or by preparation of racemic mixtures followed by resolution methods well known to those skilled in the art. These resolution methods are exemplified by (1) attachment of the mixture of enantiomers to a chiral auxiliary, separation of the resulting mixture of diastereomers by recrystallization or chromatography, and liberation of the optically pure product from the auxiliary, (2) salt formation with an optically active resolving agent, (3) direct separation of the mixture of optical enantiomers on a chiral liquid chromatography column, or (4) kinetic resolution using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be resolved into their component enantiomers by well-known methods such as chiral phase liquid chromatography or crystallization of the compound in a chiral solvent. Stereoselective synthesis, which is a chemical or enzymatic reaction in which a single reactant forms an unequal mixture of stereoisomers during the creation of a new stereocenter or the transformation of an existing stereocenter, is well known in the art. Stereoselective synthesis encompasses both enantio- and diastereoselective transformations and may involve the use of chiral auxiliaries. See, e.g., Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH: Weinheim, 2009. compound

[0094] In one embodiment, described herein is a compound represented by formula IA: [ka] or a pharma- ceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein X 1 and X 3 are each independently selected from the group consisting of CH and N; 2is NR 6 , O, and S; R 1 , R 2 , and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy and alkyl; R 5 is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acyl.

[0095] In some embodiments, R 1 , R 2 , and R 3 At least one of R is halogen. 1 , R 2 , and R 3 At least one of R is fluoro. 1 is fluoro. In some embodiments, X 1 is N.

[0096] In some embodiments, R 6 is not [ka] In the formula, L 70 is methylene or ethylene, and X 70 and X 71 One of them is CH2 and the other is N-CO-R 101 where R 101 is C 1-4 Alkyl, C 2-4 Alkenyl, or C 2-4alkynyl, each of which is unsubstituted or selected from halogen, cyano, hydroxy, amino, NH(C 1-4 alkyl), N(C 1-4 Alkyl)2, C 3-10 Cycloalkyl and C containing one or two heteroatoms each independently selected from the group consisting of nitrogen and oxygen 3-10 Heterocycloalkyl is substituted with 1 to 3 substituents each independently selected from the group consisting of heterocycloalkyl.

[0097] In another embodiment, described herein is a compound represented by formula IB: [ka] or a pharma- ceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein X 1 and X 3 are each independently selected from the group consisting of CH and N; 2 is NR 6 , O, and S; R 2 and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy and alkyl; R 5 is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acyl.

[0098] In some embodiments, X 1 is N.

[0099] In one embodiment, described herein is a compound represented by formula IC: [ka] or a pharma- ceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein X 2 is NR 6 , O, and S; X 3 is selected from the group consisting of CH and N; R 2 and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy and alkyl; R 5 is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acyl.

[0100] In some embodiments, X 2 is NR 6 It is.

[0101] In another embodiment, described herein is a compound represented by formula ID: [ka] or a pharma- ceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein R 2 and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy and alkyl; R 5is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acetate.

[0102] In some embodiments, R 2 is H and R 3 is H. In some embodiments, R 2 is F and R 3 is H. In some embodiments, R 2 is H and R 3 is F. In some embodiments, R 6 is selected from the group consisting of (C1-C8), (C2-C8)alkenyl, (C2-C8)alkenyl-(C1-C4)alkyl, (C2-C8)alkynyl, (C2-C8)alkynyl-(C1-C4)alkyl, (C3-C8)cycloalkyl, (C3-C8)cycloalkyl-(C1-C4)alkyl, (C3-C8)alkoxy-(C1-C4)alkyl, (C3-C8)cycloalkenyl, (C3-C8)cycloalkenyl-(C1-C4)alkyl, heterocyclyl, heterocyclyl-(C1-C4)alkyl, aryl, heteroaryl, and heteroaryl-(C1-C4)alkyl. In some embodiments, R 6 is selected from the group consisting of (C1-C8)alkyl, (C3-C8)cycloalkyl, (C3-C8)alkoxy-(C1-C4)alkyl, heterocyclyl, and heteroaryl. 6 is selected from the group: [ka]

[0103] In some embodiments, R 6is not [ka] In the formula, L 70 is methylene or ethylene, and X 70 and X 71 One of them is CH2 and the other is N-CO-R 101 where R 101 is C 1-4 Alkyl, C 2-4 Alkenyl, or C 2-4 alkynyl, each of which is unsubstituted or selected from halogen, cyano, hydroxy, amino, NH(C 1-4 alkyl), N(C 1-4 Alkyl)2, C 3-10 Cycloalkyl and C containing one or two heteroatoms each independently selected from the group consisting of nitrogen and oxygen 3-10 Heterocycloalkyl is substituted with 1 to 3 substituents each independently selected from the group consisting of heterocycloalkyl.

[0104] In some embodiments, R 4 is selected from the group consisting of halogen, (C1-C6)alkoxy, and C1-C6 alkyl. 4 is selected from the group consisting of chloro, fluoro, methoxy, and methyl. In some embodiments, R 5 is selected from the group consisting of H, halogen, and (C-C)alkyl. In some embodiments, R 5 is selected from the group consisting of chloro, fluoro, and methyl. In some embodiments, R 7 is H.

[0105] In some embodiments, described herein is a compound selected from the group consisting of: [ka] and the pharma- ceutically acceptable salts, enantiomers, stereoisomers, and tautomers thereof.

[0106] Treatment method The compounds described herein, e.g., compounds of formula I as defined herein, can act as therapeutic agents against diseases driven by GCN2 or PERK kinases and are useful for treating diseases and disorders in patients in need thereof, such as cancer. Exemplary cancers include colorectal cancer, rectal cancer, anal cancer, familial colorectal cancer, hereditary nonpolyposis colorectal cancer, gastrointestinal stromal tumors, lung cancer (e.g., non-small cell lung cancer, small cell lung cancer, malignant mesothelioma), mesothelioma, pancreatic cancer (e.g., pancreatic ductal carcinoma, pancreatic endocrine tumors), pharyngeal cancer, laryngeal cancer, esophageal cancer, gastric cancer (e.g., papillary adenocarcinoma, mucinous adenocarcinoma, adenosquamous carcinoma), duodenal cancer, small intestine cancer, breast cancer (e.g., invasive ductal carcinoma, non-invasive ductal carcinoma, ... in situ ductal carcinoma, inflammatory breast cancer), ovarian cancer (e.g., ovarian epithelial carcinoma, extragonadal germ cell tumor, ovarian germ cell tumor, ovarian low malignant potential tumor), testicular tumor, prostate cancer (e.g., hormone-dependent prostate cancer, non-hormone-dependent prostate cancer, castration-resistant prostate cancer), liver cancer (e.g., hepatocellular carcinoma, primary liver cancer, extrahepatic bile duct cancer), thyroid cancer (e.g., medullary thyroid carcinoma), kidney cancer (e.g., renal cell carcinoma (e.g., renal clear cell carcinoma), renal pelvis and transitional cell carcinoma of the ureter), uterine cancer (e.g., cervical cancer, uterine carcinoma, uterine sarcoma), gestational choriocarcinoma, brain tumors (e.g., medulloblastoma, glioma, pineal astrocytoma, pilocytic astrocytoma, diffuse astrocytoma, anaplastic astrocytoma, pituitary adenoma), retinoblastoma, skin cancer (e.g., basal cell carcinoma, malignant melanoma, melanoma), sarcomas (e.g., rhabdomyosarcoma, leiomyosarcoma, soft tissue sarcoma, spindle cell sarcoma, osteosarcoma), fibrosarcoma, malignant bone tumor The present invention is directed to treating or preventing cancer, bladder cancer, blood cancer (e.g., multiple myeloma, leukemia (e.g., acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), blast crisis of chronic leukemia), malignant lymphoma, Hodgkin's disease, chronic myeloproliferative disorder), cancer of unknown primary origin), cancer growth inhibitors, cancer metastasis inhibitors, apoptosis promoters, and precancerous lesions (e.g., myelodysplastic syndrome of the bone marrow).

[0107] Also described herein is a method of treating a disease caused by dysregulation of an integrated stress response in a patient in need thereof, comprising administering to the patient, in one embodiment, a therapeutically effective amount of a compound described herein (e.g., a compound of formula IA, IB, IC, and ID described herein), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the dysregulation of the integrated stress response and / or the unfolded protein response is caused by GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response and / or the unfolded protein response is caused by PERK kinase. In some embodiments, the dysregulation of the integrated stress response is caused by GCN2 kinase. In some embodiments, the dysregulation of the unfolded protein response is caused by PERK kinase. In some embodiments, the dysregulation of the integrated stress response is caused by activation of a kinase selected from the group consisting of PERK kinase and GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response is caused by activation of GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response is caused by activation of PERK kinase.

[0108] Also described herein is a method of treating a disease caused by dysregulation of the integrated stress response and / or the unfolded protein response in a patient in need thereof, comprising administering to the patient, in one embodiment, a therapeutically effective amount of a compound described herein (e.g., a compound of formula IA, IB, IC, and ID described herein), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the dysregulation of the integrated stress response and / or the unfolded protein response is caused by activation of a kinase selected from the group consisting of PERK kinase and GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response is caused by activation of GCN2 kinase. In some embodiments, the dysregulation of the unfolded protein response is caused by activation of PERK kinase.

[0109] In another embodiment, described herein is a method of modulating the activity of GCN2 kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0110] In another embodiment, described herein are methods of activating GCN2 kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0111] In another embodiment, described herein is a method of modulating activity of PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0112] In another embodiment, described herein are methods of activating PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0113] In another embodiment, described herein is a method of inhibiting GCN2 kinase and PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0114] In another embodiment, described herein is a method of modulating the activity of GCN2 kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutical acceptable salt thereof, or a pharmaceutical composition described herein.

[0115] In another embodiment, described herein are methods of inhibiting the activity of PERK kinase in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0116] In another embodiment, described herein are methods of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., compounds of formulas IA, IB, IC, and ID described herein), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, and malignant lymphoma. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, erythroleukemia, histocyctic lymphoma, Waldenstrom's macroglobulinemia, and malignant lymphoma. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is acute myeloid leukemia. In some embodiments, the cancer is acute lymphoblastic leukemia. In some embodiments, the cancer is fibrosarcoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is B-cell lymphoma. In some embodiments, the cancer is T-cell lymphoma.

[0117] In one embodiment, described herein is a method of treating amyloidosis in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein. In one embodiment, described herein is a method of treating light chain amyloidosis in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0118] In another embodiment, described herein are methods of treating a disease selected from a GCN2-related disease and a PERK-related disease in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., compounds of formulas IA, IB, IC, and ID described herein), or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition described herein. In some embodiments, the disease is a GCN2-related disease. In some embodiments, the disease is a PERK-related disease. In some embodiments, the disease is cancer. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumors, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumors, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, and malignant lymphoma. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, erythroleukemia, histiocytic lymphoma, Waldenstrom's macroglobulinemia, and malignant lymphoma. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is acute myeloid leukemia. In some embodiments, the cancer is acute lymphoblastic leukemia. In some embodiments, the cancer is fibrosarcoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is B-cell lymphoma. In some embodiments, the cancer is T-cell lymphoma.In some embodiments, the disease is amyloidosis. In some embodiments, the disease is light chain amyloidosis.

[0119] In another embodiment, described herein are methods of treating a disease selected from a GCN2-related disease and a PERK-related disease in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound described herein (e.g., compounds of formulas IA, IB, IC, and ID described herein), or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of one or more therapeutic agents. In some embodiments, the disease is a GCN2-related disease. In some embodiments, the disease is a PERK-related disease. In some embodiments, the disease is cancer. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumors, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumors, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphocytic leukemia, and malignant lymphoma. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, erythroleukemia, histiocytic lymphoma, Waldenstrom's macroglobulinemia, and malignant lymphoma. In some embodiments, the cancer is leukemia. In some embodiments, the leukemia is acute myeloid leukemia. In some embodiments, the leukemia is acute lymphocytic leukemia. In some embodiments, the cancer is fibrosarcoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is B-cell lymphoma. In some embodiments, the cancer is T-cell lymphoma.In some embodiments, the disease is amyloidosis. In some embodiments, the disease is light chain amyloidosis. In some embodiments, the one or more therapeutic agents are selected from the group consisting of L-asparaginase, pegylated asparaginase, PERK inhibitors, mTOR inhibitors, immunomodulatory agents, MAPK pathway inhibitors, MEK inhibitors, ERK inhibitors, and Ras inhibitors. In some embodiments, the one or more therapeutic agents are selected from the group consisting of IMiD agents, proteasome inhibitors, steroids, anti-CD38 agents, anti-CD20 agents, Bcl-2 inhibitors, PI3K inhibitors, bispecific antibodies, nucleoside analogs, BTK inhibitors, DNA alkylating agents, EZH2 inhibitors, anthracyclines, topoisomerase inhibitors, platins, tyrosine kinase inhibitors, HDAC inhibitors, nuclear export inhibitors, microtubule inhibitors L-asparaginase, pegylated asparaginase, PERK inhibitors, mTOR inhibitors, immunomodulatory agents, MAPK pathway inhibitors, MEK inhibitors, ERK inhibitors, and Ras inhibitors. In some embodiments, the one or more therapeutic agents are L-asparaginase, pegaspargase, calaspargase pegol-mnkl, bortezomib, carfilzomib, ixazomib, thalidomide, pomalidomide, lenalidomide, dexamethasone, prednisone, daratumumab, daratumumab / hyaluronidase, isatuximab, rituximab, obinutuzumab, venetoclax, idelalisib, copanlisib, duvelisib, umbralisib, The agent is selected from the group consisting of gemcitabine, cytarabine, ibrutinib, acalabrutinib, zanubrutinib, bendamustine, cyclophosphamide, tazemetostat, doxorubicin, daunorubicin, etoposide, oxaloplatin, carboplatin, cisplatin, bosutinib, dasatinib, imatinib, nilotinib, ponatinib, panobinostat, selinexor, vincristine, JZP-458, eriaspase, PF745 (JZP-341), asparaginase Erwinia chrysanthemi (crisantaspase), Escherichia coli asparaginase (colaspase), anti-PD1 agents, anti-PDL1 agents, and anti-CTLA4 agents.

[0120] In one embodiment, the present specification describes a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in treating a disease caused by dysregulation of an integrated stress response and / or an unfolded protein response in a patient in need of such treatment. In some embodiments, the dysregulation of the integrated stress response and / or the unfolded protein response is caused by a kinase selected from the group consisting of PERK kinase and GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response and / or the unfolded protein response is caused by GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response and / or the unfolded protein response is caused by PERK kinase. In some embodiments, the dysregulation of the integrated stress response is caused by GCN2 kinase. In some embodiments, the dysregulation of the unfolded protein response is caused by PERK kinase. In some embodiments, the dysregulation of the integrated stress response is caused by activation of a kinase selected from the group consisting of PERK kinase and GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response is caused by activation of GCN2 kinase. In some embodiments, the dysregulation of the integrated stress response is caused by the activation of PERK kinase.In one embodiment, the compounds described herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions described herein, are described herein for use in regulating the activity of GCN2 kinase in a patient in need of such regulation.In one embodiment, the compounds described herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions described herein, are described herein for use in activating GCN2 kinase in a patient in need of such regulation.In one embodiment, the compounds described herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions described herein, are described herein for use in regulating the activity of PERK kinase in a patient in need of such regulation.In one embodiment, described herein is a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in activating PERK kinase in a patient in need thereof.

[0121] In one embodiment, described herein are compounds as described herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions as described herein, for use in inhibiting GCN2 kinase and inhibiting PERK kinase in a patient in need of such inhibition. In one embodiment, described herein are compounds as described herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions as described herein, for use in modulating GCN2 kinase activity in a patient in need of such modulation. In another embodiment, described herein are compounds as described herein, or pharma- ceutically acceptable salts thereof, or pharmaceutical compositions as described herein, for use in inhibiting PERK kinase activity in a patient in need of such inhibition.

[0122] In one embodiment, described herein is a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in treating cancer in a patient in need of such treatment. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, erythroleukemia, histiocytic lymphoma, Waldenstrom's macroglobulinemia, and malignant lymphoma. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is acute myeloid leukemia. In some embodiments, the cancer is acute lymphoblastic leukemia. In some embodiments, the cancer is fibrosarcoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is B-cell lymphoma. In some embodiments, the cancer is T-cell lymphoma.

[0123] In one embodiment, described herein is a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in treating amyloidosis in a patient in need of such treatment. In one embodiment, described herein is a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in treating light chain amyloidosis in a patient in need of such treatment.

[0124] In one embodiment, described herein is a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in treating a disease selected from a GCN2-related disease and a PERK-related disease in a patient in need of such treatment. In some embodiments, the disease is a GCN2-related disease. In some embodiments, the disease is a PERK-related disease. In some embodiments, the disease is cancer. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, erythroleukemia, histiocytic lymphoma, Waldenstrom's macroglobulinemia, and malignant lymphoma. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is acute myeloid leukemia. In some embodiments, the cancer is acute lymphoblastic leukemia. In some embodiments, the cancer is fibrosarcoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is B cell lymphoma. In some embodiments, the cancer is T cell lymphoma. In some embodiments, the disease is amyloidosis. In some embodiments, the disease is light chain amyloidosis.

[0125] In one embodiment, described herein is a compound as described herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition as described herein, for use in treating a disease selected from a GCN2-related disease and a PERK-related disease in a patient in need of such treatment. In some embodiments, the disease is a GCN2-related disease. In some embodiments, the disease is a PERK-related disease. In some embodiments, the disease is cancer. In some embodiments, the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, multiple myeloma, B cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B cell lymphoma, T cell lymphoma, erythroleukemia, histiocytic lymphoma, Waldenstrom's macroglobulinemia, and malignant lymphoma. In some embodiments, the cancer is leukemia. In some embodiments, the leukemia is acute myeloid leukemia. In some embodiments, the leukemia is acute lymphoblastic leukemia. In some embodiments, the cancer is fibrosarcoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is B cell lymphoma. In some embodiments, the cancer is T cell lymphoma. In some embodiments, the disease is amyloidosis. In some embodiments, the disease is light chain amyloidosis.In some embodiments, the one or more therapeutic agents are selected from the group consisting of an IMiD agent, a proteasome inhibitor, a steroid, an anti-CD38 agent, an anti-CD20 agent, a Bcl-2 inhibitor, a PI3K inhibitor, a bispecific antibody, a nucleoside analogue, a BTK inhibitor, a DNA alkylating agent, an EZH2 inhibitor, an anthracycline, a topoisomerase inhibitor, a platin, a tyrosine kinase inhibitor, an HDAC inhibitor, a nuclear export inhibitor, a microtubule inhibitor, L-asparaginase, a pegylated asparaginase, a PERK inhibitor, an mTOR inhibitor, an immunomodulatory agent, a MAPK pathway inhibitor, a MEK inhibitor, an ERK inhibitor, and a Ras inhibitor. In some embodiments, the one or more therapeutic agents are selected from the group consisting of bortezomib, carfilzomib, ixazomib, thalidomide, pomalidomide, lenalidomide, dexamethasone, prednisone, daratumumab, daratumumab / hyaluronidase, isatuximab, rituximab, obinutuzumab, venetoclax, idelalisib, copanlisib, duvelisib, umbralisib, gemcitabine, cytarabine, ibrutinib, acalabrutinib, zanubrutinib, venetoclax ... and cisplatin, bosutinib, dasatinib, imatinib, nilotinib, ponatinib, panobinostat, selinexor, vincristine, L-asparaginase, pegaspargase, calaspargase pegol-mnkl, JZP-458, eriaspase, PF745 (JZP-341), asparaginase Erwinia chrysanthemi (crisantaspase), Escherichia coli asparaginase (colaspase), anti-PD1 agents, anti-PDL1 agents, and anti-CTLA4 agents.

[0126] The compounds provided herein can be administered to patients (animals and humans) in need of such treatment in dosages that will provide optimal pharmaceutical efficacy. It will be understood that the dosage required for use in any particular application will vary from patient to patient, depending not only on the particular compound or composition selected, but also on the route of administration, the nature of the condition being treated, the age and condition of the patient, any concomitant medications or special diets followed by the patient, and other factors that one skilled in the art would recognize, and the appropriate dosage will ultimately be left to the discretion of the attending physician. To treat the above-mentioned clinical conditions and diseases, the compounds provided herein can be administered orally, subcutaneously, topically, parenterally, by inhalation spray, or rectally in dosage unit formulations containing conventional non-toxic pharmacologic acceptable carriers, adjuvants, and vehicles. Parenteral administration can include subcutaneous injections, intravenous or intramuscular injections, or infusion techniques.

[0127] Treatment can continue for as long or as short as desired. The composition may be administered, for example, on a regimen of 1-4 times a day or more. A suitable treatment period can be, for example, at least about 1 week, at least about 2 weeks, at least about 1 month, at least about 6 months, at least about 1 year, or indefinitely. The treatment period can be terminated when the desired results are achieved.

[0128] Combination therapy The compounds described herein, e.g., compounds of formula I as defined herein, can be administered in combination with one or more additional therapeutic agents to treat the disorders described herein, such as the cancers described herein. For example, the present disclosure provides pharmaceutical compositions comprising the compounds described herein, e.g., compounds of formula I as defined herein, one or more additional therapeutic agents, and a pharma- ceutically acceptable excipient. In some embodiments, a compound of formula I as defined herein and one additional therapeutic agent are administered. In some embodiments, a compound of formula I as defined herein and two additional therapeutic agents are administered. In some embodiments, a compound of formula I as defined herein and three additional therapeutic agents are administered. Combination therapy can be achieved by administering two or more therapeutic agents, each of which is formulated and administered separately. For example, a compound of formula I as defined herein and an additional therapeutic agent can be formulated and administered separately. Combination therapy can also be achieved by administering two or more therapeutic agents in a single formulation, e.g., a pharmaceutical composition comprising a compound of formula I as one therapeutic agent and one or more additional therapeutic agents, such as a chemotherapeutic agent. For example, the compound of formula I defined herein and the additional therapeutic agent can be administered in a single formulation. Other combinations are also encompassed in combination therapy. Two or more agents in combination therapy can be administered simultaneously, but they do not have to be administered simultaneously. For example, administration of a first agent (or combination of agents) can precede administration of a second agent (or combination of agents) by minutes, hours, days, or weeks. Thus, two or more agents can be administered within minutes of each other, or within 1, 2, 3, 6, 9, 12, 15, 18, or 24 hours of each other, or within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14 days of each other, or within 2, 3, 4, 5, 6, 7, 8, 9 weeks of each other. In some cases, even longer intervals are possible. In many cases, it is desirable, but not necessary, that two or more agents used in combination therapy are present in the patient's body at the same time.

[0129] Combination therapy can also include more than one administration of one or more of the combined drugs, using a different sequence of the component drugs. For example, when drug X and drug Y are used in combination, they can be administered one or more times sequentially in any combination, such as in the sequence XYX, XXY, YXY, YYX, XXYY, etc.

[0130] Combination therapy can also include two or more administrations of one or more drugs used in combination using different administration routes.Each of one or more drugs can be administered orally, subcutaneously, topically, parenterally, by inhalation spray, or rectally, independently in a dosage unit formulation containing conventional non-toxic pharma- ceutically acceptable carriers, adjuvants, and vehicles.Parenteral administration can include subcutaneous injections, intravenous or intramuscular injections, or infusion techniques.

[0131] In some embodiments, the compounds of formula I described herein are combined with asparaginase (ASNase, L-asparaginase) or a derivative thereof. In some embodiments, the asparaginase is obtained from Erwinia chrysanthemi and is known as crisantaspase or asparaginase Erwinia chrysanthemi. Asparaginase Erwinia chrysanthemi is sold under the trademark Erwinaze® or Erwinase®. In some embodiments, the asparaginase is obtained from Escherichia coli and is known as colaspase. Colaspase is sold under the trademark Elspar®, Leunase®, Kidrolase®, or Spectrila® (recombinant E. coli aparaginase). Pegylated derivatives of colaspase are pegaspargase, sold under the trademark Oncaspar®, and calaspargase pegol-mnkl, sold under the trademark Asparlas®. Other asparaginase products currently in preclinical or clinical development include JZP-458 (recombinant Erwinia asparaginase), PF745 (JZP-341), Eryaspase (GRASPA®), and Xoncane.

[0132] In some embodiments, a compound of formula I as defined herein is combined with an immunomodulatory agent. In some embodiments, the immunomodulatory agent enhances adaptive immune responses. In some embodiments, the immunomodulatory agent enhances the activity of antigen presenting cells. In some embodiments, the immunomodulatory agent enhances the anti-tumor activity of myeloid cells, including macrophages. In some embodiments, the immunomodulatory agent enhances the anti-tumor activity of natural killer cells. In some embodiments, the immunomodulatory agent enhances the activity of effector T cells, including cytotoxic T cells.

[0133] In some embodiments, one or more additional therapeutic agents that may be administered in combination with the compounds provided herein may be MAPK pathway inhibitors, such as MEK inhibitors, ERK inhibitors, and Ras inhibitors.

[0134] Exemplary MEK inhibitors include, but are not limited to, trametinib, selumetinib, cobimetinib, binimetinib, and pharmaceutically acceptable salts thereof. Exemplary ERK inhibitors include, but are not limited to, ulixertinib, SCH772984, LY3214996, ravoxertinib, VX-11e, ASN-007, GDC-0994, MK-8353, ASTX-029, LTT462, KO-947, and pharmaceutically acceptable salts thereof. Exemplary Ras inhibitors include, but are not limited to, AMG-510, MRTX849, ARS-1620, ARS-3248, LY3499446, and pharmaceutically acceptable salts thereof.

[0135] In some embodiments, the additional therapeutic agent may be an immunomodulatory agent, including an anti-PD-1 or anti-PDL-1 therapeutic, including, but not limited to, pembrolizumab, nivolumab, pidilizumab, cemiplimab, atezolizumab, durvalumab, BMS-936559, or avelumab. In some embodiments, the additional therapeutic agent may be an anti-TIM3 (anti-HAVcr2) therapeutic agent, including but not limited to TSR-022 or MBG453, an anti-LAG3 therapeutic agent, including but not limited to leratolimab, LAG525, or TSR-033, an anti-4-1BB (anti-CD37, anti-TNFRSF9), a CD40 agonist therapeutic agent, including but not limited to SGN-40, CP-870,893, or RO7009789, an anti-CD47 therapeutic agent, including but not limited to Hu5F9-G4, an anti-CD20 therapeutic agent, an anti-CD38 therapeutic agent, a STING agonist, including but not limited to ADU-S100, MK-1454, ASA404, or an amide benzimidazole. In some embodiments, the additional therapeutic agent may be an anti-CTLA4 agent, including ipilimumab, tremelimumab. In some embodiments, the additional therapeutic agent may be a hypomethylating agent, including but not limited to azacitidine or decitabine, epidermal growth factor inhibitors, statins, metformin, angiotensin receptor blockers, other immunomodulatory therapies, including but not limited to thalidomide, lenalidomide, pomalidomide, prednisone, or dexamethasone. In some embodiments, the additional therapeutic agent may be an immunotherapeutic agent, including targeted therapeutics, cancer vaccines, and CAR-T cell therapies.

[0136] The compounds of formula I described herein may be administered in combination with other therapeutic agents known to treat cancer, including radiation therapy, antitubulin agents, DNA alkylating agents, DNA synthesis inhibitors, DNA intercalating agents, antiestrogens, antiandrogens, steroids, anti-EGFR drugs, kinase inhibitors, mTOR inhibitors, PI3 kinase inhibitors, cyclin-dependent kinase inhibitors, CD4 / CD6 kinase inhibitors, topoisomerase inhibitors, histone deacetylase (HDAC) inhibitors, DNA methylation inhibitors, anti-HER2 agents, antiangiogenic agents, proteasome inhibitors, PARP (poly ADP-ribose polymerase) inhibitors, cell cycle regulating kinase inhibitors, thalidomide, lenalidomide, pomalidomide, bortezomib, carfilzomib, ixazomib, daratumumab, daratumumab / hyaluronidase, isotuximab, dexamethasone, and antibody-drug-conjugates (ADCs).

[0137] In one embodiment, the additional therapeutic agent is an antitubulin agent (e.g., paclitaxel, paclitaxel protein-bound particles for injectable suspension including nab-paclitaxel, eribulin, docetaxel, ixabepilone, vincristine, auristatin, or maytansinoids), vinorelbine, DNA alkylating agents (including cisplatin, carboplatin, oxaliplatin, cyclophosphamide, ifosfamide, temozolomide), DNA intercalating agents or DNA topoisomerase inhibitors (including, for example, anthracyclines such as doxorubicin, pegylated liposomal doxorubicin, daunorubicin, idarubicin, mitoxantrone, or epirubicin; camptothecins such as topotecan, irinotecan, or exatecan), 5-fluorouracil, capecitabine, cytarabine, decitabine, 5-azacitadine (5-azacitadine), The chemotherapy agents may include cyclosporine, gemcitabine, and methotrexate.

[0138] In some embodiments, the additional therapeutic agent may be a kinase inhibitor, including but not limited to erlotinib, gefitinib, neratinib, afatinib, osimertinib, lapatanib, crizotinib, brigatinib, ceritinib, alectinib, lorlatinib, everolimus, temsirolimus, abemaciclib, LEE011, palbociclib, cabozantinib, ripretinib, sunitinib, pazopanib, sorafenib, regorafenib, sunitinib, axitinib, dasatinib, imatinib, nilotinib, idelalisib, ibrutinib, BLU-667, loxo 292, larotrectinib, and quizartinib.

[0139] In some embodiments, the additional therapeutic agent is an anti-estrogen, including but not limited to tamoxifen, fulvestrant, anastrozole, letrozole, and exemestane; an anti-androgen, including but not limited to abiraterone acetate, enzalutamide, nilutamide, bicalutamide, flutamide, cyproterone acetate; a steroid, including but not limited to prednisone and dexamethasone; a PARP inhibitor, including but not limited to neraparib, olaparib, talazoparib, and rucaparib; a topoisomerase inhibitor, including but not limited to irinotecan, camptothecin, exatecan, and topotecan. The inhibitor may be a biologic agent including, but not limited to, tetracycline I inhibitors, topoisomerase II inhibitors including, but not limited to, anthracyclines, etoposide, etoposide phosphate, and mitoxantrone, histone deacetylase (HDAC) inhibitors including, but not limited to, vorinostat, romidepsin, panobinostat, valproic acid, and belinostat, DNA methylation inhibitors including, but not limited to, DZNep and 5-aza-2'-deoxycytidine, proteasome inhibitors including, but not limited to, bortezomib and carfilzomib, trastuzumab, adotrastuzumab, pertuzumab, cetuximab, and panitumumab.

[0140] In some embodiments, the additional therapeutic agent can be an anti-angiogenic agent, including bevacizumab, aflibercept, and AMG386.

[0141] In some embodiments, the additional therapeutic agent may be a therapeutic vaccine, including but not limited to, DM1, DM4, MMAE, MMAF, or antibody-drug-conjugates (ADCs) containing a payload of camptothecin, brentuximab vedotin and trastuzumab emtansine, radiation therapy, sipuleucel-T.

[0142] In some embodiments, the additional therapeutic agent may be an autophagy inhibitor, including a ULK inhibitor, a VPS34 inhibitor, a PIKfyve inhibitor, a PPT1 inhibitor, or a lysosomal blocker. In some embodiments, the additional therapeutic agent may be DCC-3116, SAR405, SB02024, hydroxychloroquinine, chloroquinine, apilimod, MRT403, and LYS05.

[0143] In some embodiments, the additional therapeutic agent is selected from luteinizing hormone releasing hormone (LHRH) analogs, including goserelin and leuprolide.

[0144] In some embodiments, the additional therapeutic agent is everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, AZD 1152, enzastaurin, vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, pemetrexed, erlotinib, dasatanib, nilotinib, decatanib, panitumumab, amrubicin, oregovomab, Lep-etu, nolatrexed, AZD 2171, butabulin, ofatumtunab, zanolimumab, edotecarin, tetrandrine, rubitecan, tesmifene, oblimersen, ticilimumab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, cilengitide, gymatecan, IL13-PE38QQR, INO 1001, IPdR1KRX-0402, lucanton, LY 317615, neuradiab, vitespan, Rta 744, alanosine (Sdx 102), talampanel, atrasentan, XR 311, romidepsin, ADS-100380, sunitinib, 5-fluorouracil, vorinostat, etoposide, gemcitabine, doxorubicin, irinotecan, liposomal doxorubicin, 5'-deoxy-5-fluorouridine, vincristine, temozolomide, ZK-304709, seliciclib;PD0325901, AZD-6244, capecitabine, L-glutamic acid, N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidin-5-yl)-ethyl]benzoyl]-, disodium salt, heptahydrate, camptothecin, PEG-labeled irinotecan, tamoxifen, toremifene citrate, anastrozole, exemestane, letrozole, DES (diethylstilbestrol), estradiol, estrogen, conjugated estrogen, bevacizumab, IMC-1C11, CHIR-258, 3-[5-(methylsulfonylpiperazinemethyl)-indolyl]-quinolone, vatalanib, AG-013736, AVE-0005, [D-Ser(tBu)6, Azgly 10] acetate (Pyro-Glu-His-Trp-Ser-Tyr-D-Ser(tBu)-Leu-Arg-Pro-Azgly-NH2 acetate [C; 59 H 84 N 18 O 14 -(C2H4O2) x(wherein x=1-2.4)], goserelin acetate, leuprolide acetate, triptorelin pamoate, medroxyprogesterone acetate, hydroxyprogesterone caproate, megestrol acetate, raloxifene, bicalutamide, furutanide, nilutamide, megestrol acetate, CP-724714; TAK-165, HKI-272, erlotinib, lapatanib, canertinib, ABX-EGF antibody, Erbitux, EKB-569, PKI-166, GW-572016, ionafarnib, BMS-214662, tipifarnib; amifostine, NVP-LAQ824, suberoylanilide hydroxamic acid, valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, amsacrine, anagrelide, L-asparaginase, Bacillus Calmette-Guerin (BCG) vaccine, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol , epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gemcitabine, Gleevec, hydroxyurea, idarubicin, ifosfamide, imatinib, leuprolide, levamisole, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, mitomycin, mitotane, mitoxantrone, nilutamide, octreotide, oxaliplatin, pamidronate, pentostatin, plicamycin, porfimer, procarbazine, raltitrexed, rituximab, streptomycin, Tozosin, teniposide, testosterone, thalidomide, thioguanine, thiotepa, tretinoin, vindesine, 13-cis-retinoic acid, phenylalanine mustard, uracil mustard, estramustine, altretamine, floxuridine, 5-deoxyuridine, cytosine arabinoside, 6-mecaptopurine, deoxycoformycin, calcitriol, valrubicin, mithramycin, vinblastine, vinorelbine, topotecan, razoxine, marimastat, COL-3, neovastat, BMS-275291, squalamine,Endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifene, idoxifene, spironolactone, finasteride, cimitidine, trastuzumab, denileukin diftitox, gefitinib, bortezimib, irinotecan, topotecan, doxorubicin, docetaxel, vinorelbine, bevacizumab (monoclon lonal antibody) and Erbitux, Cremophor-free paclitaxel, Epithilon B, BMS-247550, BMS-310705, droloxifene, 4-hydroxytamoxifen, pipendoxifene, ERA-923, arzoxifene, fulvestrant, acolbifene, lasofoxifene, idoxifene, TSE-424, HMR-3339, ZK186619, PTK787 / ZK 222584, VX-745, PD 184352, rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, temsirolimus, AP-23573, RAD001, ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, wortmannin, ZM336372, L-779450, PEG-filgrastim, darbepoetin, erythropoietin, granulocyte-colony stimulating factor, zoledronate, prednisone, cetuximab, granulocyte-macrophage colony-stimulating factor, histrelin, pegylated interferon alpha-2a, interferon alpha-2a, pegylated interferon alpha-2b, interferon Alpha-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-trans retinoic acid, ketoconazole, interleukin-2, megestrol, immunoglobulin, nitrogen mustard, methylprednisolone, ibritumomab tiuxetan, androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, editronate, mitotane, cyclosporine, liposomal daunorubicin, edwina-asparaginase, strontium 89, casopitant, netupitant, NK1 receptor blockers,Selected from the group consisting of palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, dolasetron, tropisetron, pegfilgrastim, erythropoietin, epoetin alfa and darbepoetin alfa, ipilumumab, and mixtures thereof.

[0145] Pharmaceutical Compositions and Kits Another aspect of the present disclosure provides pharmaceutical compositions comprising the compounds disclosed herein formulated together with a pharma- ceutically acceptable carrier. Specifically, the present disclosure provides pharmaceutical compositions comprising the compounds disclosed herein formulated together with one or more pharma- ceutically acceptable carriers. These formulations include those suitable for oral, rectal, topical, buccal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous) rectal, vaginal, or aerosol administration, although the most suitable administration form in any given case depends on the extent and severity of the condition being treated and the nature of the particular compound being used. For example, the disclosed compositions may be formulated as a unit dose and / or may be formulated for oral or subcutaneous administration.

[0146] Exemplary pharmaceutical compositions can be used in the form of pharmaceutical preparations, for example, in solid, semi-solid, or liquid form, containing one or more of the compounds described herein as active ingredients mixed with organic or inorganic carriers or excipients suitable for external, enteral, or parenteral application. The active ingredients can be compounded with conventional non-toxic pharmaceutically acceptable carriers for, for example, tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions, and any other form suitable for use. The subject active compounds are included in the pharmaceutical composition in an amount sufficient to produce the desired effect on the disease process or condition.

[0147] To prepare solid compositions such as tablets, the primary active ingredient can be mixed with pharmaceutical carriers, such as conventional tableting ingredients, such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gums, and other pharmaceutical diluents, such as water, to form solid preformulation compositions containing a homogenous mixture of the compounds provided herein, or non-toxic pharma- ceutically acceptable salts thereof. When these preformulation compositions are referred to as homogenous, it is meant that the active ingredient is evenly dispersed throughout the composition, such that the composition may be readily subdivided into equally effective unit dosage forms, such as tablets, pills, and capsules.

[0148] In solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.), the subject compositions are mixed with one or more pharma- ceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and / or acacia; (3) humectants, such as glycerol; (4) disintegrants, such as glycerol; Disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (5) solution retarders such as paraffin, (6) absorption accelerators such as quaternary ammonium compounds, (7) wetting agents such as acetyl alcohol and glycerol monostearate, (8) absorbents such as kaolin and bentonite clay, (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof, and (10) coloring agents. In the case of capsules, tablets, and pills, the compositions may also contain buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols, and the like.

[0149] Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binders (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethylcellulose), surface active agents, or dispersants. Molded tablets may be made by molding a mixture of the subject composition moistened with an inert liquid diluent in a suitable machine. Tablets, as well as other solid dosage forms such as dragees, capsules, pills, and granules, may be optionally scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art.

[0150] Compositions for inhalation or insufflation include solutions and suspensions in pharma- ceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders.Liquid dosage forms for oral administration include pharma-ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.In addition to the subject compositions, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters, cyclodextrins, and mixtures thereof.

[0151] Suspending agents may include, in addition to the subject compositions, suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, and mixtures thereof.

[0152] Formulations for rectal or vaginal administration may be prepared by mixing the subject compositions with one or more suitable non-irritating excipients or carriers including, for example, cocoa butter, polyethylene glycol, a suppository wax, or a salicylate, and may be presented as a suppository, which is solid at room temperature but becomes liquid at body temperature and therefore melts in the body cavity releasing the active agent.

[0153] Dosage forms for transdermal administration of the subject compositions include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active component may be mixed under sterile conditions with a pharma- ceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.

[0154] The ointments, pastes, creams, and gels may contain, in addition to the subject compositions, excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0155] Powders and sprays may contain, in addition to the subject composition, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0156] Alternatively, the compositions and compounds of the present disclosure may be administered by aerosol. This is accomplished by preparing an aqueous aerosol, liposomal preparation, or solid particles containing the compounds. A non-aqueous (e.g., fluorocarbon propellant) suspension may be used. An ultrasonic nebulizer may be used because it minimizes exposure of the drug to shear, which may result in degradation of the compounds contained in the subject compositions. Typically, aqueous aerosols are made by formulating an aqueous solution or suspension of the subject compositions with conventional pharma- ceutically acceptable carriers and stabilizers. Carriers and stabilizers vary according to the requirements of a particular subject composition, but typically include non-ionic surfactants (Tween, Pluronic®, or polyethylene glycol), innocuous proteins such as serum albumin, sorbitan esters, amino acids such as oleic acid, lecithin, glycine, buffers, salts, sugars, or sugar alcohols. Aerosols are generally prepared from isotonic solutions.

[0157] Pharmaceutical compositions of the present disclosure suitable for parenteral administration comprise the subject compositions in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately prior to use, and may contain antioxidants, buffers, bacteriostats, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents.

[0158] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions provided herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate and cyclodextrin. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0159] In another embodiment, an enteral pharmaceutical formulation is provided that includes the disclosed compound and an enteric material, and a pharma- ceutically acceptable carrier or excipient thereof. An enteric material refers to a polymer that is substantially insoluble in the acidic environment of the stomach and is predominantly soluble in intestinal fluids at a particular pH. The small intestine is the part of the digestive tract (intestine) between the stomach and the large intestine, and includes the duodenum, jejunum, and ileum. The pH of the duodenum is about 5.5, the pH of the jejunum is about 6.5, and the pH of the distal ileum is about 7.5.

[0160] Thus, the enteric material will not dissolve until the pH is, for example, about 5.0, about 5.2, about 5.4, about 5.6, about 5.8, about 6.0, about 6.2, about 6.4, about 6.6, about 6.8, about 7.0, about 7.2, about 7.4, about 7.6, about 7.8, about 8.0, about 8.2, about 8.4, about 8.6, about 8.8, about 9.0, about 9.2, about 9.4, about 9.6, about 9.8, or about 10.0. Exemplary enteric materials include cellulose acetate phthalate (CAP), hydroxypropyl methylcellulose phthalate (HPMCP), polyvinyl acetate phthalate (PVAP), hydroxypropyl methylcellulose acetate succinate (HPMCAS), cellulose acetate trimellitate, hydroxypropyl methylcellulose succinate, cellulose acetate succinate, cellulose acetate hexahydrophthalate, cellulose propionate phthalate, cellulose acetate maleate, cellulose acetate butyrate, cellulose acetate propionate, copolymers of methyl methacrylate and methyl methacrylate, copolymers of methyl acrylate, copolymers of methyl methacrylate and methacrylic acid, copolymers of methyl vinyl ether and maleic anhydride (Gantrez ES series), ethyl methacrylate-methyl methacrylate-chlorotrimethylammonium ethyl acrylate copolymers, natural resins such as zein, shellac, and copalcohol, as well as several commercially available enteric dispersions (e.g., Eudragit L30D55, Eudragit FS30D, Eudragit Examples of suitable enteric materials include enteric esters such as enteric esters L100, Eudragit S100, Kollicoat EMM30D, Estacryl 30D, Coateric, and Aquateric. The solubility of each of the above materials is either known or readily determinable in vitro. While the foregoing is a list of possible materials, one of ordinary skill in the art having the benefit of this disclosure will recognize that it is not comprehensive and that there are other enteric materials that will meet the objectives described herein.

[0161] Advantageously, kits are provided herein for use by a purchaser in need of, for example, a cancer treatment. Such kits include appropriate dosage forms, such as those described above, and instructions that explain how to use such dosage forms to mediate, reduce, or prevent inflammation. The instructions direct the consumer or medical practitioner to administer the dosage forms according to modes of administration known to those skilled in the art. Such kits may advantageously be packaged and sold in single or multiple kit units. One example of such a kit is the so-called blister pack. Blister packs are well known in the packaging industry and are widely used for packaging pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister packs generally consist of a sheet of relatively stiff material covered with a foil of a preferably transparent plastic material. During the packaging process, recesses are formed in the plastic foil. These recesses have the size and shape of the tablets or capsules to be packed. The tablets or capsules are then placed in the recesses and the sheet of relatively stiff material is sealed against the plastic foil with the face of the foil opposite to the direction in which the recesses were formed. As a result, the tablets or capsules are sealed in the recesses between the plastic foil and the sheet. Preferably, the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by manually applying pressure to the recesses, thereby forming openings in the sheet at the locations of the recesses, through which the tablets or capsules can then be removed.

[0162] It may be desirable to provide a memory aid on the kit, for example in the form of numbers next to the tablets or capsules, which numbers correspond to the days of the regimen on which the tablets or capsules so designated should be taken. Another example of such a memory aid is a calendar printed on a card, for example, "Week 1, Monday, Tuesday, ... etc. ... Week 2, Monday, Tuesday, ...", etc. Other variations of memory aids will be readily apparent. A "daily dose" can be a single tablet or capsule or multiple tablets or capsules to be taken on a given day. Also, a daily dose of a first compound can consist of one tablet or capsule and a daily dose of a second compound can consist of multiple tablets or capsules, and vice versa. The memory aid should reflect this. EXAMPLES

[0163] The compounds described herein can be prepared in a number of ways based on the teachings contained herein and the disclosure of synthetic procedures in the art. In the description of the synthesis method, Selection of solvent, reaction atmosphere, reaction temperature, All suggested reaction conditions, including experimental durations and work-up procedures, are within the scope of the present invention unless otherwise stated. The conditions can be selected to be the standard for the reaction. The functionality present on various parts of the molecule It is understood that the substituents must be compatible with the proposed reagents and reactions. Substituents that are not compatible with the reaction conditions are It will be apparent to those skilled in the art that alternative methods may be used. Starting materials for the examples are either commercially available or can be prepared from known sources. It is easily prepared by standard methods.

[0164] The following abbreviations are used in this disclosure and have the following definitions: "AcOH" is acetic acid, "ADP" is adenosine diphosphate, "Ar" is argon gas, "ASNase" is asparaginase, "Boc" is t-butyl carbonate, and "conc" is acetylated ketone. "Cs2CO3" is cesium carbonate, "DCM" is dichloromethane, "DIAD" is diisopropyl azodicarboxylate, "DIEA" is N,N-diisopropylethylamine, "DMF" is N,N-dimethylformamide, "dppf" is 1,1'-bis(diphenylphosphino)ferrocene, "DMSO" is dimethylsulfoxide, "EDTA" is ethylenediaminetetraacetic acid, "ESI" is electrospray ionization, "EtOAc" is ethyl acetate, "EtOH" is ethanol, "GST" is glutathione S-transferase, "h" is time (hour or hours), "HCl" is hydrochloric acid, "Hex" is hexane, "H2O" is water, and "IC 50" is the 50% inhibitory concentration, "K2CO3" is potassium carbonate, "KOAc" is potassium acetate, "NaBH4" is sodium borohydride, "LAH" is lithium aluminum hydride, "CH3CN" is acetonitrile, "mCPBA" is 4-chloroperbenzoic acid, "MeOH" is methanol, "MHz" is megahertz, "min" is minute or minutes, "MS" is mass spectrometry, "NADH" is nicotinamide adenine dinucleotide, "NaH" is sodium hydride, "NaHCO3" is sodium bicarbonate, "NaNO2" is sodium nitrite, "NaOMe" is sodium methoxide, and "Na2SO4" is sodium sulfate. , "NBS" is N-bromosuccinimide, "NCS" is N-chlorosuccinimide, "NH4Cl" is ammonium chloride, "NIS" is N-iodosuccinimide, "NMR" is nuclear magnetic resonance, "PBS" is phosphate buffered saline, "Pd2(dba)3" is tris(dibenzylideneacetone)dipalladium(0), Pd(dppf)Cl2" is 1,1-bis(diphenylphosphino)ferrocene-palladium(II) dichloride, "Ph3P" is triphenylphosphine, "PMB" is paramethoxybenzyl, "POCl3" is phosphorus oxychloride, "rt" is room temperature, also known as ambient temperature, understood to consist of the range of standard laboratory temperatures ranging from 15 to 25°C, and "sat'd" is is saturated, "SDS" is sodium dodecyl sulfate, "SOCl2" is thionyl chloride, "TEA" is triethylamine, "TFA" is trifluoroacetic acid, "THF" is tetrahydrofuran, and "XantPhos" is 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene. general chemistry

[0165] Exemplary compounds described herein are accessible by the general synthetic methods illustrated in the following schemes, intermediate preparations, and accompanying examples. [ka]

[0166] Scheme 1 shows an exemplary preparation of sulfonyl chloride 1.5. Bromide 1.1a (commercially available or synthesized by one skilled in the art) is converted to thioether 1.2a by Pd-catalyzed coupling reaction (e.g., using Pd2(dba)3, XantPhos, phenylmethanethiol in a solvent such as toluene at elevated temperature in the presence of a base such as DIEA). Alternatively, compound 1.2 can be prepared from aniline 1.1b (commercially available or synthesized by one skilled in the art) by diazotization using dibenzyl disulfide and amyl nitrite in CH3CN at elevated temperature. Reduction of ester 1.2 (R = Me, Et) with a reducing agent (LAH or NaBH4) gives primary alcohol 1.3. Alcohol 1.3 can be converted to 1.4 by reaction with acetic anhydride in the presence of a base such as DIEA. Finally, smooth oxidation of thioether 1.4 with a combination of NCS and AcOH affords the corresponding sulfonyl chloride 1.5, following the general reaction conditions reported in Synthesis, 2006, 24, 4131-4134 and Bioorg. Med. Chem., 2017, 25, 3447-3460. [ka]

[0167] Scheme 2 shows an exemplary preparation of boronate 2.4. Compound 2.1 (commercially available, synthesized as described in WO2013 / 134298, or synthesized by one skilled in the art) is reacted with bis(pinacolato)diboron by a boronation reaction known to those skilled in the art (palladium-mediated reaction carried out at elevated temperature in a suitable solvent such as 1,4-dioxane using a palladium catalyst such as Pd(dppf)Cl2 and a suitable base such as KOAc) to give compound 2.2, which is reacted with sulfonyl chloride 1.5 (see Scheme 1) to give compound 2.4. Alternatively, compound 2.1 is reacted with sulfonyl chloride 1.5 to give sulfonamide 2.3, which is converted to boronate 2.4 under boronation reaction conditions known to those skilled in the art. [ka]

[0168] Scheme 3 shows an exemplary preparation of boronate 3.3. Compound 3.2 is prepared from aniline 3.1 (either commercially available or synthesized by one skilled in the art) by diazotization followed by Cu-mediated chlorination of the resulting intermediate (according to the general reaction conditions reported in Org. Proc. Res. Dev., 2009, 5, 875-879). Sulfonyl chloride 3.2 is reacted with amine 2.2 under sulfonamide coupling to give sulfonamide 3.3. [ka]

[0169] Scheme 4 shows an exemplary preparation of intermediate 4.4. In the presence of a base such as NaH, an alkylating agent (R 5 N-alkylation of 3-iodo-1H-pyrrolo[3,2-c]pyridine 4.1 with 0)2SO2 gives 4.2. Compound 4.2 can be activated by art-known oxidation such as mCPBA in DCM to give N-oxide 4.3. Pyrrolopyridine N-oxide 4.3 is converted to aminopyrrolopyridine 4.4 using a mixture of PMBNH2 and p-toluenesulfonyl chloride, followed by in situ deprotection with TFA. [ka]

[0170] Scheme 5 shows an exemplary preparation of intermediate 5.8. Various amines (R 5Cyclization of 2-(4,6-dichloropyrimidin-5-yl)acetaldehyde 5.1 with NBS (or NIS) affords compound 5.2. Bromination (or iodination) of 5.2 with NBS (or NIS) affords compound 5.4 (X=CH). Reaction of compound 5.5 with NBS (or NIS) affords compound 5.6. Compounds 5.3, 5.5, and 5.6 can be reacted with an alkylating agent (R 5 Alkylation with 5.4, 5.7, and 5.8, respectively, can be obtained by alkylation with 5.4, 5.7, and 5.8 under standard Mitsunobu conditions. 5 -OH) (e.g., performed in the presence of Ph3P and DIAD) to give 5.4, 5.7, and 5.8, respectively. In another embodiment, intermediate 5.8 can also be prepared from 5.4 by displacement reaction with ammonium hydroxide, and from 5.7 by bromination (or iodination) with NBS (or NIS). [ka]

[0171] Scheme 6 shows an exemplary preparation of formula I. Bromide (or iodide) 4.4 and 5.8 react with boronate 2.2 in the presence of a palladium catalyst (Suzuki conditions) to give aniline 6.1. Sulfonamide coupling reaction of aniline 6.1 with sulfonyl chloride 1.5 gives sulfonamide 6.3. Alternatively, 6.3 can be prepared from bromide (or iodide) 4.4 and 5.8 with boronate 2.4 under Suzuki conditions. Deprotection of sulfonamide 6.3 with K2CO3 in a protic solvent such as MeOH at room temperature gives formula I. In another embodiment, bromide (or iodide) 4.4 and 5.8 react with boronate 3.3 in the presence of a palladium catalyst (Suzuki conditions) to give 6.2. Reduction of 6.2 under LiAlH4 gives primary alcohol, formula I. Preparation of intermediates and final compounds.

[0172] Using the synthetic procedures and methods described herein, as well as methods known to one of ordinary skill in the art, the following compounds were made: General Method A: Borylation Example A1: 2-Fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline [ka]

[0173] A mixture of KOAc (4.6 g, 47.4 mmol), 3-bromo-2-fluoroaniline (3.0 g, 15.8 mmol), and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (4.8 g, 18.9 mmol) in 1,4-dioxane (30 mL) was degassed with Ar for 10 min. PdCl2(dppf) (1.3 g, 1.6 mmol) was added and the reaction mixture was heated at 110 °C for 6 h. The reaction was cooled to room temperature and filtered through a pad of Celite. The filtrate was removed under reduced pressure and the residue was purified by silica gel column chromatography (0-50% EtOAc / Hexanes) to give 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (2.7 g, 72% yield) as a tan solid. 1 H NMR(500 MHz,DMSO-d6):δ 6.86(m,2H),6.76(d,J=6.8 Hz,1H),5.02(s,2H),1.28(s,12H);MS(ESI)m / z:238.2(M+H + ).

[0174] Using General Method A above, the intermediates in Table A below were prepared. [Table A]

[0175] General Method B: Alkylation: Example B1: 4-Chloro-5-iodo-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine [ka] A solution of 4-chloro-5-iodo-7H-pyrrolo[2,3-d]pyrimidine (2.0 g, 7.2 mmol) in DMF (18 mL) was treated with Cs2CO3 (4.7 g, 14 mmol). 2-Iodopropane (1.2 g, 7.2 mmol) was added and the reaction mixture was stirred at room temperature for 12 h. The reaction was poured into water (100 mL) and the solid was collected via vacuum filtration to give 4-chloro-5-iodo-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine (1.8 g, 78%) as a yellow fluffy solid. 1 H NMR(500 MHz,DMSO-d6):δ 8.63(s,1H),8.18(d,J=1.4 Hz,1H),5.04(m,1H),1.47(d,J=6.8 Hz,6H);MS(ESI)m / z:322.0(M+H + ).

[0176] Using general method B above, the intermediates in Table B below were prepared. [Table B]

[0177] Preparation of B6: (E)-N'-(5-iodo-7-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide [ka]

[0178] A solution of 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (5.0 g, 19 mmol) in DMF (50 mL) was treated with N,N-dimethylformamide dimethyl acetal (3.43 g, 28 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure and the residue was diluted with DCM (150 mL) and washed with brine (2×) solution. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The crude material was triturated with diethyl ether (100 mL) and the solid was filtered to give the desired (E)-N'-(5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide (5.0 g, 85%). 1 H NMR(500 MHz,DMSO-d6):δ 11.9(s,1H),8.80(s,1H),8.27(s,1H),7.41(d,J=2.0,1H),3.21(s,3H),3.17(s,3H); MS(ESI)m / z:315.8(M+H + ).

[0179] A solution of (E)-N'-(5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide (2.0 g, 6.3 mmol) and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.6 g, 7.6 mol) in DMF (30 mL) under oxygen atmosphere was treated with NaHCO3 (1.3 g, 12.6 mmol), 2,2-bipyridyl (1.1 g, 6.9 mmol) and Cu(OAc)2 (1.3 g, 6.9 mol) at room temperature. The reaction mixture was stirred at room temperature for 4 days and concentrated under reduced pressure. The crude material was triturated with diethyl ether (60 mL) and the solid was filtered to give the desired (E)-N'-(5-iodo-7-(1-methyl-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide (3.0 g, crude), which was used in the next reaction without further purification. MS (ESI) m / z: 395.8 (M+H + ). Example B7: (E)-N'-(5-iodo-7-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide [ka]

[0180] (E)-N'-(5-iodo-7-(1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide was prepared from (E)-N'-(5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-N,N-dimethylformimidamide (0.69 g, 2.2 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole pyrazole (0.5 g, 2.6 mol) analogously to Example B6. MS (ESI) m / z: 381.8 (M+H + ).

[0181] General method C: cyclization Example C1: 4-Chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine [ka] A solution of 2-(4,6-dichloropyrimidin-5-yl)acetaldehyde (2.0 g, 10.5 mmol) in EtOH (18 mL) was treated with Et3N (2.9 mL, 21 mmol). The reaction mixture was stirred at room temperature for 10 min, then cyclopropanamine (0.6 mL, 9.4 mmol) was added. The reaction mixture was stirred at 140° C. overnight. The reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0-100% EtOAc / Hexanes) to give 4-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine (1.6 g, 79%) as a pale yellow solid. 1H NMR(500 MHz,DMSO-d6):δ 8.59(s,1H),7.65(d,J=3.7 Hz,1H),6.54(d,J=3.5 Hz,1H),3.59(m,1H),1.01-1.03(m,4H); MS(ESI)m / z:194.0(M+H + ).

[0182] Using general procedure C above, the intermediates in Table C below were prepared. [Table C]

[0183] General Method D: Bromination or Iodination Example D1: 5-Bromo-4-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine [ka] A solution of 4-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine (C1, 0.67 g, 3.5 mmol) in DCM (15 mL) was treated with NBS (0.74 g, 4.2 mmol) and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure and the residue was suspended in water (5 mL). The solid was filtered, washed with water (2 mL) and dried under vacuum to give 5-bromo-4-chloro-7-cyclopropyl-7H-pyrrolo[2,3-d]pyrimidine (0.6 g, 64%) as a white solid. 1 H NMR(500MHz,DMSO-d6):δ8.69(s,1H),8.00(d,J=2.1Hz,1H),3.66(m,1H),1.10(m,4H);MS(ESI)m / z:272.0(M+H + ) and 274.

[0184] Using General Method D above, the intermediates in Table D below were prepared. [Table D]

[0185] General method E: Substitution Example E1: 5-Iodo-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine [ka] A suspension of 4-chloro-5-iodo-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine (B1, 1.78 g, 5.5 mmol) in 1,4-dioxane (5.5 mL) was treated with NH4OH (30% in water, 1.7 mL, 13 mmol) and the reaction was heated to 95° C. overnight. The mixture was cooled to room temperature and diluted with cold water. The mixture was stirred at 0° C. for 30 minutes, then the solid was collected via vacuum filtration to give the desired product 5-iodo-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (0.80 g, 48%) as a white solid. 1 H NMR(500 MHz,DMSO-d6):δ 8.09(s,1H),7.59(s,1H),6.59(s,2H),4.89(m,1H),1.40(d,J=6.8 Hz,6H); MS(ESI)m / z:303.0(M+H + ).

[0186] Using general procedure E above, the intermediates in Table E below were prepared. [Table E-1] [Table E-2]

[0187] Example E12: Preparation of 3-iodo-1-methyl-1H-pyrrolo[3,2-c]pyridin-4-amine [ka] A stirred suspension of 3-iodo-1-methyl-1H-pyrrolo[3,2-c]pyridine (0.90 g, 3.5 mmol) in DCM (20 mL) was treated with 3-chloroperoxybenzoic acid (0.90 g, 5.2 mmol) at room temperature. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with saturated NaHCO3 and then the solution was extracted with DCM (2x). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give crude 3-iodo-1-methyl-1H-pyrrolo[3,2-c]pyridine 5-oxide (0.6 g, 65%) as an off-white solid. 1 H NMR(400 MHz,DMSO-d6):δ 8.12(s,1H),8.00(d,J=7.2 Hz,1H),7.34(s,1H),7.59(d,J=7.2 Hz,1H),3.82(s,3H); MS(ESI)m / z:275.0(M+H) + ).

[0188] A solution of 3-iodo-1-methyl-1H-pyrrolo[3,2-c]pyridine 5-oxide (0.6 g, 2.1 mmol) in chloroform (20 mL) was treated with (4-methoxyphenyl)methanamine (1.3 g, 9.8 mmol) and stirred at room temperature for about 30 min. The mixture was cooled to 0° C., followed by the addition of p-toluenesulfonyl chloride (0.92 g, 4.8 mmol) in small portions and the reaction mixture was stirred at room temperature for 2.5 h. The mixture was diluted with DCM (50 mL) and washed with saturated NaHCO3 solution. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude material was purified by silica gel column chromatography (70% EtOAc / Hexanes) to give 3-iodo-N-(4-methoxybenzyl)-1-methyl-1H-pyrrolo[3,2-c]pyridin-4-amine (0.2 g, 23%) as a pale yellow solid. 1H NMR(400 MHz,DMSO-d6):δ 7.68(d,J=6.4 Hz,1H),7.36(s,1H),7.31(d,J=7.6 Hz,2H),6.89(d,J=7.6 Hz,2H),6.81(d,J=6.0 Hz,1H),6.14(br m,1H),4.64(d,J=5.2 Hz,2H),3.73(s,3H),3.69(s,3H); MS(ESI)m / z:394.2(M+H + ).

[0189] A suspension of 3-iodo-N-(4-methoxybenzyl)-1-methyl-1H-pyrrolo[3,2-c]pyridin-4-amine (0.2 g, 0.508 mmol) in DCM (10 mL) was stirred under ice-water bath. TFA (1 mL) was then added dropwise and the reaction mixture was stirred at room temperature for 4 h. The reaction mixture was concentrated under reduced pressure and the residue was treated with a saturated solution of NaHCO3. The mixture was stirred for 10 min and the aqueous layer was further extracted with 10% MeOH / DCM (2x). The combined organic extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (3-5% MeOH / DCM) to give 3-iodo-1-methyl-1H-pyrrolo[3,2-c]pyridin-4-amine (0.05 g, 36%) as an off-white solid. 1 H NMR(400 MHz,DMSO-d6):δ 7.63,d,J=6.4 Hz,1H),7.48(s,1H),6.94(d,J=6.4 Hz,1H),6.35(br s,2H),3.72(s,3H); MS(ESI)m / z:273.9(M+H) + ).

[0190] General Method F: Preparation of Thioethers from Amines Example F1: Ethyl 3-(benzylthio)-2,5-dichlorobenzoate [ka]

[0191] A solution of ethyl 3-amino-2,5-dichlorobenzoate (12.0 g, 51.0 mmol) in CH3CN (250 mL) was treated with amyl nitrite (9.64 mL, 81.0 mmol). Dibenzyl disulfide (12.6 g, 51.0 mmol) was added at room temperature and the reaction mixture was heated at 70 °C for 3 h. The reaction mixture was quenched with ice water (100 mL) and extracted with EtOAc (3x). The combined organics were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (10-20% EtOAc / Hexanes) to give ethyl 3-(benzylthio)-2,5-dichlorobenzoate (10.0 g, 58% yield) as an off-white solid. 1 H NMR(500 MHz,DMSO-d6):δ 7.63(d,J=2.8 Hz,1H),7.56(d,J=2.4 Hz,1H),7.44(d,J=7.2 Hz,2H),7.35(t,J=7.6 Hz,2H)7.28(d,J=7.2 Hz,1H),4.42(s,2H),4.33(q,J=7.2 Hz,2H),1.29(t,J=6.8 Hz,3H).

[0192] General Method G: Preparation of Thioethers from Bromine Example F2: Ethyl 3-(benzylthio)-2-chlorobenzoate [ka]

[0193] A solution of methyl 3-bromo-2-chlorobenzoate (3.0 g, 12 mmol), phenylmethanethiol (1.6 g, 13 mmol) in toluene (30 mL) was treated with DIEA (4.2 mL, 24 mmol). The mixture was purged with Ar for 5 min, then XantPhos (0.67 g, 1.2 mmol) and Pd2(dba)3 (0.55 g, 0.60 mmol) were added. The mixture was purged with Ar for 5 min and heated to 90 °C overnight. The reaction was cooled to room temperature, filtered through silica gel, and washed with EtOAc:Hexanes (1:1). The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (0-100% EtOAc:Hexanes) to give methyl 3-(benzylthio)-2-chlorobenzoate (2.8 g, 80%) as an orange oil. 1 H NMR(500 MHz,DMSO-d6):δ 7.60(dd,J=8.0,1.5 Hz,1H),7.50(dd,J=7.6,1.5 Hz,1H),7.37-7.46(m,2H),7.31-7.37(m,2H),7.23-7.31(m,2H),4.34(s,2H),3.85(s,3H).

[0194] Using general procedures F and G above, the intermediates in Table F below were prepared. [Table F]

[0195] General method H: reduction Example G1: (3-(benzylthio)-2,5-dichlorophenyl)methanol [ka]

[0196] A mixture of CaCl2 (0.45 g, 0.92 mmol) and NaBH4 (6.20 g, 37 mmol) in EtOH (200 mL) was stirred at 0 °C. A cold solution of ethyl 3-(benzylthio)-2,5-dichlorobenzoate (F1, 14.0 g, 9.2 mmol) in THF (200 mL) was added dropwise at 0 °C. The reaction mixture was stirred at 0 °C, slowly warmed to room temperature, and then heated at 60 °C for 5 h. The reaction mixture was cooled to room temperature, quenched with saturated NH4Cl solution (50 mL), and then extracted with EtOAc (3x). The combined organic extracts were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (1-10% EtOAc / Hexanes) to give (3-(benzylthio)-2,5-dichlorophenyl)methanol (8.0 g, 66%) as a brown solid. 1 H NMR(400 MHz,DMSO-d6):δ 7.44(m,2H),7.35(m,3H),7.25-7.32(m,2H),5.57(t,J=5.6 Hz,1H),4.51(d,J=6.0 Hz,2H),4.36(s,2H).

[0197] Using general procedure H above, the intermediates in Table G below were prepared. [Table G]

[0198] General Method I: Acetylation Example H1: 3-(benzylthio)-2,5-dichlorobenzyl acetate [ka]

[0199] A solution of (3-(benzylthio)-2,5-dichlorophenyl)methanol (G1, 0.82 g, 2.7 mmol) in THF (10 mL) was treated with acetic anhydride (0.31 mL, 3.3 mmol) at room temperature. The mixture was stirred at 50 °C under Ar for 2 h and then diluted with water. The solution was extracted with EtOAc (3x) and the combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 3-(benzylthio)-2,5-dichlorobenzyl acetate (0.92 g, 98%) as a white solid. 1 H NMR (400 MHz, DMSO-d6): δ 7.51(m,3H),7.41(m,3H),7.35(m,1H),5.16(s,2H),4.45(s,2H),2.16(s,3H).

[0200] Using general method I above, the intermediates in Table H below were prepared. [Table H-1] [Table H-2]

[0201] General Method J: Preparation of Sulfonyl Chlorides Example I1: 2,5-Dichloro-3-(chlorosulfonyl)benzyl acetate [ka]

[0202] A solution of 3-(benzylthio)-2,5-dichlorobenzyl acetate (H1, 0.92 g, 2.7 mmol) and 1-chloropyrrolidine-2,5-dione (1.8 g, 13 mmol) in THF / H2O (1:1, 3 mL) at 0 °C was treated with acetic acid (7 mL). The reaction mixture was allowed to warm slowly to room temperature and then stirred at room temperature for 6 h. The reaction mixture was quenched with saturated NaHCO3 solution (100 mL) and extracted with EtOAc (3x). The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0-100% EtOAc / Hexanes) to give 2,5-dichloro-3-(chlorosulfonyl)benzyl acetate (0.45 g, 53%) as a white solid. 1 H NMR (400 MHz, DMSO-d6): δ 7.84(d,J=2.8 Hz,1H),7.52(d,J=2.8 Hz,1H),5.13(s,2H),2.11(s,3H).

[0203] Using general procedure J above, the intermediates in Table I below were prepared. [Table I-1] [Table I-2]

[0204] General Method K: Sulfonamide Coupling Example J1: 2,5-dichloro-3-(N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfamoyl)benzoic acid methyl ester [ka]

[0205] A solution of 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (A12.2 g, 9.3 mmol) in DCM (30 mL) was treated with pyridine (2.2 mL, 28 mmol). The reaction mixture was cooled to 0° C. and a solution of methyl 2,5-dichloro-3-(chlorosulfonyl)benzoate (I22.8 g, 9.3 mmol) in DCM (10 mL) was added dropwise. The reaction mixture was allowed to warm slowly to room temperature over 2 h. The reaction mixture was concentrated under reduced pressure and the crude was dissolved in DCM (50 mL). The solution was washed with 1.0 M HCl (2×), aqueous NaHCO3 (3×), and brine (1×). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was suspended in hexane and the solid was collected by vacuum filtration to give methyl 2,5-dichloro-3-(N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfamoyl)benzoate (4.2 g, 90%) as a brown solid. 1 H NMR(500 MHz,DMSO-d6):δ 10.71(s,1H),8.13(d,J=2.6 Hz,1H),8.02(d,J=2.6 Hz,1H),7.46(m,1H),7.38(td,J=7.9,1.8 Hz,1H),7.16(t,J=7.7 Hz,1H),3.91(s,3H),1.28(s,12H); MS(ESI)m / z 526.0(M+Na+H + ).

[0206] General method L: reduction Example J2: Preparation of 2,5-dichloro-N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(hydroxymethyl)benzenesulfonamide [ka]

[0207] A solution of methyl 2,5-dichloro-3-(N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfamoyl)benzoate (J1, 0.5 g, 0.99 mmol) in THF (5 mL) was treated portionwise with LAH (0.11 g, 30 mmol) at 0° C. and then stirred at 0° C. for 1 h. The reaction mixture was diluted with diethyl ether and then quenched with water (0.2 mL), 15% aqueous NaOH (0.2 mL), and water (0.4 mL). The mixture was stirred for 4 h and then filtered through a pad of Celite. The filtrate was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 2,5-dichloro-N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3-(hydroxymethyl)benzenesulfonamide (0.26 g, 55%) as a colorless solid, which was used in the next reaction without further purification. MS (ESI) m / z 498.0 (M+Na+H + ).

[0208] Using general procedures K and L above, the intermediates in Table J below were prepared. [Table J-1] [Table J-2]

[0209] General Method M: Suzuki Reaction Example 1: 3-(N-(3-(4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)sulfamoyl)-2,5-dichlorobenzyl acetate [ka]

[0210] A solution of 5-iodo-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.7 g, 6.3 mmol) in a mixture of 1,4-dioxane and water (4:1, 100 mL) was stirred under Ar. 2,5-dichloro-3-(N-(2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfamoyl)benzyl acetate (3.6 g, 6.9 mmol) and Cs2CO3 (5.1 g, 15.7 mmol) were added at room temperature and the reaction mixture was degassed with Ar for 5 min. Pd(dppf)Cl2.DCM (0.25 g, 0.31 mmol) was added and the resulting mixture was heated at 90 °C for 1.5 h. The reaction was cooled to room temperature and filtered through a pad of Celite, washing with EtOAc. The filtrate was washed with saturated NaHCO3 (2x) followed by water (2x). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the crude material. The crude material was further stirred in CH3CN (300 mL) at room temperature for 2 h. The solid was filtered and dried under vacuum to give 3-(N-(3-(4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)sulfamoyl)-2,5-dichlorobenzyl acetate (2.5 g, 91%) as a light brown solid. 1 H NMR(400 MHz,DMSO-d6):δ 10.76(br s,1H),8.14(s,1H),7.93(d,J=2.4 Hz,1H),7.84(m,1H),7.26(s,1H),7.19(m,3H),6.02(br s,2H),5.20(s,2H),3.73(s,3H),3.56(s,3H); MS(ESI)m / z 538.0(M+H + ).

[0211] General method N: Deprotection Example 2: N-(3-(4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-2,5-dichloro-3-(hydroxymethyl)benzenesulfonamide [ka]

[0212] A solution of 3-(N-(3-(4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)sulfamoyl)-2,5-dichlorobenzyl acetate (1, 2.3 g, 4.2 mmol) in MeOH (40 mL) was treated with K2CO3 (2.3 g, 17 mmol) at room temperature. The mixture was stirred at room temperature for 3 h and then the solvent was evaporated under reduced pressure. The crude material was acidified with 10% citric acid solution (pH approx. 5) and then the solid was filtered and washed thoroughly with water. The solid was treated with a mixture of CH3CN (5 mL) and DCM (5 mL) at room temperature for 2 h. The precipitate was filtered and dried to give N-(3-(4-amino-7-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-2-fluorophenyl)-2,5-dichloro-3-(hydroxymethyl)benzenesulfonamide (1.6 g, 76%) as an off-white solid. 1 H NMR(400 MHz,DMSO-d6)δ 10.69(br s,1H),8.14(s,1H),7.84(d,J=2.4 Hz,1H),7.78(br m,1H),7.27(s,1H),7.15-7.25(br m,3H),5.99(br s,2H),5.72(t,J=5.6 Hz,1H),4.60(d,J=5.6 Hz,2H),3.73(s,3H).MS(ESI)m / z 496.27(M+H + ).

[0213] Using general methods L, M and N above, the intermediates in Table K below were prepared. [Table K-1] [Table K-2] [Table K-3] [Table K-4] [Table K-5] [Table K-6]

[0214] Example 30. Biochemical Assays of GCN2 GCN2 kinase activity was determined using a TR-FRET kinase activity assay (e.g., Riddle et al. Analytical Biochemistry (2006) 356(1) 108-116). Assays were performed in 384-well plates (13 uL assay volume) using 2 nM GCN2 (Carna Biosciences), 130 nM GFP-EIf2a (Invitrogen), 0.2 mg / mL E. coli tRNA (sigma), and 1 mM ATP (Invitrogen) in kinase buffer. Inhibition of GCN2 was measured by adding serially diluted test compounds (final assay concentration 0.5% DMSO) followed by a 3 hour incubation. Tb-peIF2a (pSer52) antibody (Invitrogen) (final assay concentration 2 nM) in kinase buffer containing ETDA (final assay concentration 20 mM) was added. After 60 min incubation at room temperature, TR-FRET was monitored using an excitation wavelength of 340 nm and emission wavelengths of 490 nm and 520 nm. The emission ratio (520 / 490) at each compound concentration was converted to percent inhibition using controls (i.e., reactions without test compound and reactions with known inhibitors) and calculated as IC 50 Values ​​were calculated by fitting a four parameter sigmoidal curve to the data using Prism (GraphPad software).

[0215] SEQ ID NO:1 - GCN2 protein sequence (residues 1-1649, G556E with N-terminal GST tag)

[0216] Example 31. Biochemical assay of PERK PERK kinase activity was determined spectrophotometrically using a coupled pyruvate kinase / lactate dehydrogenase assay (e.g., Schindler et al. Science (2000) 289:1938-1942), which continuously monitors the ATP hydrolysis-dependent oxidation of NADH. Assays were performed in 384-well plates (final volume 100 μL) using 10 nM PERK (Beryllium), 0.25 mg / mL myelin basic protein, 1.5 units of pyruvate kinase, 2.1 units of lactate dehydrogenase, 1 mM phosphoenolpyruvate, 0.28 mM NADH, and 1 mM ATP in assay buffer (100 mM Tris, pH 7.5, 15 mM MgCl2, 0.5 mM DTT, 0.004% (w / v) BSA, and 0.004% Triton X-100). Inhibition of PERK was measured by adding serially diluted test compounds (final assay concentration 1% DMSO). The decrease in absorbance at 340 nm was monitored continuously for 6 h at 30 °C on a multimode microplate reader (BioTek). Reaction rates were calculated using a 2-3 h time frame. Reaction rates at each concentration of compound were converted to percent inhibition using controls (i.e., reactions without test compound and reactions with known inhibitors) and calculated as IC 50 Values ​​were calculated using software routines in Prism (GraphPad software).

[0217] SEQ ID NO:2 - PERK protein sequence (residues 563-1115, SEQ ID: NM — 004836) MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLK SSKYIAWPLQGWQATFGGGDHPPKSDLVPRGSKYDSVSGEANDSSWNDIKNSGYISRYLTDFEPIQCLGRGGFGVVFEAKNKVDDCNYAIKRIRLPNRELAREKVMREVKALAKLEHPGIVRYFNAWLEAPPEKWQEKMDEIWLKDESTDWPLSSPSPMDAPSVKIRRMDPFSTKEHIEIIAPSPQRSRSFSVGI SCDQTSSSESQFSPLEFSGMDHEDISESVDAAYNLQDSCLTDCDVEDGTMDGNDEGHSFELCPSEASPYVRSRERTSSSIVFEDSGCDNASSKEEPKTNRLHIGNHCANKLTAFKPTSSKSSSEATLSISPPRPTTLSLDLTKNTTEKLQPSSPKVYLYIQMQLCRKENLKDWMNGRCTIEERERSVCLHIFLQI AEAVEFLHSKGLMHRDLKPSNIFFTMDDVVKVGDFGLVTAMDQDEEEQTVLTPMPAYARHTGQVGTKLYMSPEQIHGNSYSHKVDIFSLGLILFELLYPFSTQMERVRTLTDVRNLKFPPLFTQKYPCEYVMVQDMLSSPMERPEAINIIENAVFEDLDFPGKTVLRQRRSRSLSSSGTKHSRQSNNSHSPLPSN [Table 1]

[0218] In Table 1, "+" indicates IC<100 nM 50 "++" indicates IC > 100nM and < 500nM 50 "+++" indicates IC greater than 500nM and less than 1000nM50 "++++" indicates IC greater than 1000nM and less than 10000nM 50 Refers to...

[0219] In the following examples and figures provided herein, "Compound 2" refers to the compound of Example 2 above. Example 32. A phenotypic assay for cellular inhibition of GCN2, the CCRF-CEM ASNase cell proliferation assay.

[0220] CCRF-CEM leukemia cells (catalog number CCL-116) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cells were grown in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA) and 1% penicillin / streptomycin / L-glutamine at 37°C, 5% CO2 and 95% humidity. Cells were grown until they reached 1 million cells per mL, at which point they were subcultured or harvested for assay use. Ten thousand cells per well in 200 μL of RPMI-1640 supplemented with 10% heat-inactivated fetal bovine serum and 1% penicillin / streptomycin were dispensed into 96-well black clear-bottom plates. Serial dilutions of test compounds and 1 mU / mL ASNase were added in triplicate and the plates were incubated for 72 hours at 37°C, 5% CO2, and 95% humidity. At the end of the incubation, 40 μL of a 440 mM resazurin (Sigma, St. Louis, MO) solution in PBS was added to each well of the plate and the plates were incubated for an additional 6 hours at 37°C, 5% CO2, and 95% humidity. Plates were read on a Synergy2 (Biotek, Winooski, VT) or equivalent reader using an excitation of 540 nm and an emission of 600 nm. Data were analyzed using GraphPad Prism software (GraphPad, San Diego, CA) to determine IC 50 The value was calculated. Example 33. HCT116 Amino Acid Starved (-AA) Phospho-GCN2 and ATF4 Assays

[0221] HCT116 colorectal cancer cells (catalog number CCL-247) were obtained from American Type Culture Collect (ATTC, Manassas, VA). Briefly, cells were grown in RPMI1640 supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA) and 1% penicillin / streptomycin / L-glutamine at 37°C, 5% CO2, and 95% humidity. Cells were expanded until they reached 70-95% confluency, at which point they were subcultured or harvested for assay use. Cells were seeded into 12-well culture plates at 500,000 cells per well in 1 mL of complete growth medium and incubated overnight at 37°C, 5% CO2, and 95% humidity. The next day, each well was replaced with 1 mL of Earle's Balanced Salt Solution (EBSS, Invitrogen, Carlsbad, CA) supplemented with 10% dialyzed fetal bovine serum, 5.5 mM glucose, 1% penicillin / streptomycin, and 1% vitamin solution (thermo catalog number 11120052). Serial dilutions of test compounds were dispensed into the wells. The plates were incubated for 4 hours at 37°C, 5% CO2, and 95% humidity. At the end of the incubation, the cells were washed with PBS supplemented with 1X Halt protease inhibitor, 1X Halt phosphatase inhibitor, 1X Sigma phosphatase inhibitor cocktail 2, and 1X EDTA, and then lysed with M-PER Mammalian Protein Extraction Reagent supplemented with 3X inhibitor mix as described above. The cell lysate was sonicated in a water bath sonicator (Qsonica, Newtown, CT), and the supernatant was boiled with SDS buffer and reducing agent. Western blots were performed on each lysate to quantify phosphorylated GCN2 (Thr899), total GCN2, ATF4, and β-actin. Membranes were imaged using a LI-COR Odyssey CLx Imaging System (LI-COR, Lincoln, NE). Data were analyzed using GraphPad Prism software (GraphPad, San Diego, CA) to determine IC 50 The value was calculated. Example 34. A phenotypic assay for cellular inhibition of pre-activated PERK by thapsigargin (TG): CCRF-CEM TG ATF4 ELISA

[0222] CCRF-CEM leukemia cells (catalog number CCL-116) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Briefly, cells were grown in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA) and 1% penicillin / streptomycin / L-glutamine at 37°C, 5% CO2, and 95% humidity. Cells were grown until they reached 1 million cells per mL, at which point they were subcultured or harvested for assay use. 1.5 million cells per well in 1 mL of complete growth medium were dispensed into 12-well plates and incubated overnight. Serial dilutions of test compounds were added and cells were incubated at 37°C, 5% CO2, and 95% for 3 hours, then 1 μμM thapsigargin was added and cells were incubated at 37°C, 5% CO2, and 95% for an additional hour. Cells were lysed and then ATF4 levels were measured using an ELISA assay (Proteintech, Rosemont, IL). Absorbance was measured at 450 nM and 544 nM using a Synergy2 or equivalent reader (Biotek, Winooski, VT). Data was analyzed using Prism software (GraphPad, San Diego, CA) to determine IC 50 The value was calculated. [Table 2]

[0223] In Table 2, "+" indicates IC<100 nM 50 "++" indicates IC > 100nM and < 500nM 50 "+++" indicates IC greater than 500nM and less than 1000nM 50"++++" indicates IC greater than 1000nM and less than 10000nM 50 Refers to... Example 35. H929 ATF4 ELISA Assay

[0224] H929 multiple myeloma cells (catalog number CRL-9068) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Briefly, cells were grown in RPMI-1640 medium supplemented with 20% heat-inactivated fetal bovine serum (catalog number A3840002, ThermoFisher Scientific, Waltham, MA), 1% penicillin / streptomycin / L-glutamine (catalog number 10378016, ThermoFisher Scientific, Waltham, MA), and 0.05 mM 2-mercaptoethanol (catalog number 21985-023, ThermoFisher Scientific, Waltham, MA) at 37°C, 5% CO2, and 95% humidity. Cells were grown until they reached 1.5 million cells per mL, at which point they were either subcultured or harvested for assay use. 1.5 million cells per well in 1 mL of complete growth medium were dispensed into 12-well plates and incubated overnight. Serial dilutions of test compounds were added and cells were incubated for 4 hours at 37°C, 5% CO2 and 95% CO2. Cells were lysed and then ATF4 levels were measured using an ELISA assay (Proteintech, Rosemont, IL). Absorbance was measured at 450 nM and 544 nM using a Synergy2 or equivalent reader (Biotek, Winooski, VT). Data was analyzed using PRISM software (Graphpad, San Diego, CA) to calculate fold stimulation of cellular ATF4 relative to vehicle-treated controls. [Table 3]

[0225] In Table 3, "+" refers to stimulation of ATF4 of 5-fold or less, "++" refers to stimulation of ATF4 of more than 5-fold and less than 10-fold, and "+++" refers to stimulation of ATF4 of more than 10-fold and less than 20-fold.

[0226] Figure 1 is a graphical representation showing the unexpected stimulation of the UPR / ISR marker ATF4 (black bars) in response to increasing concentrations of compound 2. Stimulation occurred in a bell-shaped manner, with ATF4 levels found to peak at concentrations ranging from 4 to 123 nM, a concentration range in which 50% or less of PERK molecules were occupied by compound 2, followed by an inhibitory phase in which ATF4 expression returned to basal levels at concentrations of 370 nM or greater of compound 2. Maximal stimulation in the ATF4 signal was 13-fold and occurred at approximately 41 nM compound 2. Example 36. Detection of PERK oligomerization using NanoBRET

[0227] Constructs for BRET were constructed using the NanoBRET™ Flexi® PPI Starter System (Promega, Madison, WI). Full-length PERK ORF was obtained from Genscript, Inc. (Piscataway, NJ). HEK-293 cells (catalog no. CRL-1573) were obtained from American Type Culture Collection (ATCC, Manassas, VA). Cells were grown in MEM medium supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA) and 1% penicillin / streptomycin / L-glutamine at 37° C., 5% CO2, and 95% humidity. For BRET assays, cells were seeded at a density of 40,000 cells / mL in culture medium in 6-well plates and allowed to attach and recover. Cells were transfected with C-terminally tagged PERK-NLuc and C-terminally tagged PERK-Halo using Lipofectamine LTX (Thermo, Waltham, MA) and proteins were expressed overnight at 37°C, 5% CO2, and 95% humidity. Cells were then detached from the culture dish using 0.05% trypsin EDTA, collected, and diluted to 22,000 cells / mL in assay medium consisting of Opti-MEM® I reduced serum medium supplemented with 4% FBS (Thermo). Cells were dispensed into 96-well cell culture plates and HaloTag® NanoBRET™ 618 Ligand (Promega) was added to a final concentration of 100 nM. Compounds or DMSO were added to the cells and the plates were incubated for 4 hours at 37°C, 5% CO2, and 95% humidity. NanoBRET™ Nano-Glo® Substrate was added to the cells and the cells were shaken for 30 seconds. Donor (460 nM) and acceptor (618) emissions were measured within 10 minutes of substrate addition on a Synergy Neo2 multimode reader (BioTek, Winooski, VT). Data was reported as BRET ratio (acceptor / donor).

[0228] Compound 2 has an EC 50 and unexpectedly exhibited stimulation of PERK oligomer formation. A graph of the stimulated PERK oligomer formation induced by compound 2 is shown in FIG. Example 37. Stimulation of UPR / ISR signaling pathway proteins ATF4 and CHOP in multiple myeloma

[0229] H929 multiple myeloma cells (catalog no. CRL-9068) were obtained from the American Type Culture Collection (ATTC, Manassas, VA) and grown and maintained at 1-2 million cells per mL in RPMI 1640 supplemented with 20% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA), 1% penicillin / streptomycin / L-glutamine, and 0.05 mM 2-mercaptoethanol at 37°C, 5% CO2, and 95% humidity. Cells were seeded at 4 million cells per well in 2 mL of complete growth medium in 6-well plates and treated with serial dilutions of test compounds for 4 hours at 37°C, 5% CO2, and 95% humidity. At the end of the incubation, cells were washed with PBS supplemented with 1X Halt protease inhibitor, 1X Halt phosphatase inhibitor, 1X Sigma phosphatase inhibitor cocktail 2, and 1X EDTA, then lysed with M-PER mammalian protein extraction reagent supplemented with 3X inhibitor mix, as described above. Cell lysates were sonicated in a water bath sonicator (Qsonica, Newtown, CT), and supernatants were boiled with SDS buffer and reducing agent. Western blots were performed to quantify ATF4, CHOP, and beta-actin (Cell Signaling Technology, Danvers, MA). Membranes were imaged with a LI-COR Odyssey CLx Imaging System (LI-COR, Lincoln, NE).

[0230] Compound 2 unexpectedly exhibited stimulation of the PERK pathway in H929 multiple myeloma cells. A diagram of stimulated PERK downstream signaling proteins ATF4 and CHOP (with actin as a loading control) is shown in FIG. 3. ATF4 is stimulated over a concentration range of 4.5 nM to 123 nM. CHOP is stimulated over a concentration range of 4.5 nM to 41 nM. Example 38. Stimulation of UPR / ISR target genes in multiple myeloma using quantitative RT-PCR

[0231] H929 multiple myeloma cells (catalog no. CRL-9068) were obtained from the American Type Culture Collection (ATTC, Manassas, VA). Cells were cultured at 37°C, 5% CO 2、 Cells were grown in RPMI-1640 medium supplemented with 20% heat-inactivated fetal bovine serum (catalog no. A3840002, ThermoFisher Scientific, Waltham, MA), 1% penicillin / streptomycin / L-glutamine (catalog no. 10378016, ThermoFisher Scientific, Waltham, MA), and 0.05 mM 2-mercaptoethanol (catalog no. 21985-023, ThermoFisher Scientific, Waltham, MA) at 4°C and 95% humidity. Cells were grown until they reached 70-95% confluence, at which point they were either subcultured or harvested for assay use. Cells were plated at 3.0 × 10 per well in 2 mL of complete growth medium in 6-well culture plates. 6Cells were seeded and incubated with the indicated concentrations of Compound 2 for 4 hours at 37°C, 5% CO2, and 95% humidity. At the end of the incubation, cells were washed with PBS (Sigma) and RNA was extracted using the RNeasy® Plus kit (Qiagen, Germantown, MD). cDNA was synthesized using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosciences, Beverly Hills, CA) and quantitative PCR was performed using TaqMan Assays (Table 1, Thermo) and TaqMan Fast Advanced Master Mix (Thermo) in a QuantStudio 3 (Applied Biosciences) according to the manufacturer's specifications. [Table 4]

[0232] Compound 2 unexpectedly induced the expression of UPR / ISR target genes in H929 multiple myeloma cells. Data was reported as fold change relative to vehicle control (DMSO) treated samples. Figure 4 is a graphical representation showing the stimulation of ATF4 target genes in response to increasing concentrations of compound 2 (DMSO black bar, 1.5 nM dark gray bar, 14 nM checkered bar, 41 nM striped bar, 123 nM light gray bar, and 370 nM dotted bar). Transcriptional activation of ATF4 target genes occurred in a bell-shaped manner. The maximum stimulation of ATF4 at 4 hours was about 3-fold, the maximum stimulation of CHOP at 4 hours was about 10-fold, the maximum stimulation of GADD34 at 4 hours was about 3-fold, the maximum stimulation of GPT2 at 4 hours was about 3-fold, and the maximum stimulation of VEGFA at 4 hours was about 2-fold. Stimulation of ATF4 target genes was found to occur in a bell-shaped manner, with levels peaking at concentrations ranging from 14 to 123 nM, a concentration range in which 50% or less of PERK molecules were occupied by compound 2, followed by an inhibitory phase in which expression returned to basal levels at concentrations of compound 2 ≥ 370 nM. Example 39. Stimulation of multiple myeloma apoptosis signaling pathways

[0233] H929 multiple myeloma cells (catalog no. CRL-9068) were obtained from the American Type Culture Collection (ATTC, Manassas, VA) and grown and maintained at 1-2 million cells per mL in RPMI 1640 supplemented with 20% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA), 1% penicillin / streptomycin / L-glutamine, and 0.05 mM 2-mercaptoethanol at 37°C, 5% CO2, and 95% humidity. Cells were seeded at 4 million cells per well in 2 mL of complete growth medium in 6-well plates and treated with serial dilutions of test compounds for 24 hours at 37°C, 5% CO2, and 95% humidity. At the end of the incubation, cells were washed with PBS supplemented with 1X Halt protease inhibitor, 1X Halt phosphatase inhibitor, 1X Sigma phosphatase inhibitor cocktail 2, and 1X EDTA, then lysed with M-PER mammalian protein extraction reagent supplemented with 3X inhibitor mix, as described above. Cell lysates were sonicated in a water bath sonicator (Qsonica, Newtown, CT), and supernatants were boiled with SDS buffer and reducing agent. Western blots were performed to quantify cleaved-caspase 3, cleaved-caspase 7, PARP, and beta-actin (Cell Signaling Technology, Danvers, MA). Membranes were imaged with a LI-COR Odyssey CLx Imaging System (LI-COR, Lincoln, NE).

[0234] Compound 2 unexpectedly induced the expression of pro-apoptotic proteins in H929 multiple myeloma cells. Treatment of cells with compound 2 resulted in increased levels of cleaved PARP, cleaved caspase-7, and cleaved caspase-3. Stimulation of these apoptotic readouts occurred in a bell-shaped fashion, peaking at concentrations ranging from 4.5 nM to 41 nM. Example 40. Inhibition of Multiple Myeloma and B-cell Lymphoma Cell Proliferation

[0235] H929, RPMI8226, GA-10, and DoHH-2 cells (catalog no. CRL-9068) were obtained from the American Type Culture Collection (ATTC, Manassas, VA). H929 cells were maintained in RPMI-1640 medium supplemented with 20% heat-inactivated fetal bovine serum (catalog no. A3840002, ThermoFisher Scientific, Waltham, MA), 1% penicillin / streptomycin / L-glutamine (catalog no. 10378016, ThermoFisher Scientific, Waltham, MA), and 0.05 mM 2-mercaptoethanol (catalog no. 21985-023, ThermoFisher Scientific, Waltham, MA). RPMI8226, GA-10, DoHH-2 were maintained in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (catalog no. A3840002, ThermoFisher Scientific, Waltham, MA) and 1% penicillin / streptomycin / L-glutamine (catalog no. 10378016, ThermoFisher Scientific, Waltham, MA). All cells were grown at 37°C, 5% CO2, and 95% humidity. Cells were grown until they reached 1.5 million cells per mL, at which point they were either subcultured or harvested for assay use.

[0236] For cell proliferation assays, appropriate numbers of cells (40,000 cells for H929; 20,000 cells for RPMI8226 and GA-10; 8,000 cells for DoHH-2) were seeded in 200 μL of each medium per well and dispensed into 96-well black clear-bottom plates. Serial dilutions of test compounds were added in triplicate and plates were incubated at 37° C., 5% CO2, and 95% humidity for 72 or 120 hours. Several standard of care agents were used in the combination studies (lenalidomide (catalog no. S1029, Selleckchem, Houston, TX), bortezomib (catalog no. S1013, Selleckchem, Houston, TX), dexamethasone (catalog no. S1322, Selleckchem, Houston, TX), ibrutinib (Pharmacyclics, Sunnyvale, CA). At the end of the incubation in H929, RPMI8226, and GA-10, 440 mM resazurin in PBS was added. 40 μL of (Sigma, St. Louis, MO) solution was added to each well of the plate and the plate was incubated for an additional 7 hours at 37°C, 5% CO2, and 95% humidity. Plates were read in a Synergy2 or equivalent reader (Biotek, Winooski, VT) using excitation at 540 nm and emission at 600 nm. At the end of the DoHH-2 incubation, cell viability was determined using the CellTiter-Glo® assay (Promega, Madison, WI). Luminescence was measured using an EnVision MultiLabel Reader (PerkinElmer, Waltham, MA). Data was analyzed using GraphPad Prism software (GraphPad, San Diego, CA) to determine IC 50 The value was calculated.

[0237] Compound 2 shows additive or synergistic effects on inhibition of cell proliferation when combined with standard of care (SOC) agents for treating multiple myeloma or B-cell lymphoma. Representative graphs are shown in Figures 6A, 6B, 6C, and 6D. Figure 6A represents a cell proliferation experiment of RPMI8226 multiple myeloma cells treated with compound 2 titers with (grey diamonds) or without (filled circles) 333 nM dexamethasone for 96 hours. The combined treatment resulted in inhibition of cell proliferation with an IC50 of 7 nM. Figure 6B represents a cell proliferation experiment of H929 multiple myeloma cells treated with compound 2 titers with (grey squares) or without (filled circles) 3 nM bortezomib for 96 hours. The combined treatment resulted in inhibition of cell proliferation with an IC50 of 6 nM. Figure 6C represents a cell proliferation experiment of GA-10 Burkitt's lymphoma cells treated with compound 2 potencies with (grey triangles) or without (filled circles) 50 nM dexamethasone for 96 hours. The combined treatment resulted in inhibition of cell proliferation with an IC50 of 4 nM. Figure 6D represents a cell proliferation experiment of follicular lymphoma cells treated with compound 2 potencies with (grey stars) or without (filled circles) 41 nM ibrutinib for 96 hours. The combined treatment resulted in inhibition of cell proliferation with an IC50 of 4 nM. 50 resulted in inhibition of cell proliferation. Example 41. CCRF-CEM xenograft pharmacokinetic / pharmacodynamic (PK / PD) model

[0238] The CCRF-CEM xenograft model was performed in an AAALAC-accredited facility in accordance with all laws, regulations, and guidelines of the National Institutes of Health (NIH) and with approval from the Animal Care and Use Committee at Labcorp (Ann Arbor, MI). Food and water were provided ad libitum. All mice were observed for clinical signs at least once daily. Female Envigo CB-17 SCIDs (6-7 weeks old) were inoculated subcutaneously just below the right upper axilla with 10 million cells in Dulbecco's phosphate-buffered saline using a 27-gauge needle and syringe. Tumor burden averaged 200 mm on day 22. 3When tumor size reached 100 μg / kg, mice were randomly assigned to groups such that the mean tumor burden for all groups was within 10% of the overall mean tumor burden for the study population. Groups were treated on days 23-24 as follows: vehicle control (administered orally and IP to mimic the combination group) (n=9) on day 23; ASNase (Abcam, Cambridge, UK) administered IP at 1000 U / kg / day (n=9). On day 24, subjects were randomly assigned to vehicle control (administered orally and IP to mimic the combination group) (n=9); Compound 2 orally at 50 mg / kg / day Compound 2 (n=9) and ASNase (Abcam, Cambridge, UK) IP at 1000 U / kg / day 2 hours prior to sample collection (n=9); Compound 2 orally at 25 mg / kg / day Compound 2 (n=9) and ASNase (Abcam, Cambridge, UK) IP at 1000 U / kg / day 2 hours prior to sample collection at 2, 6, and 10 hours after Compound 2 administration (n=9). Blood samples were collected in K2EDTA tubes, processed to plasma, flash frozen in liquid nitrogen, and then stored at -80°C. Plasma samples were subjected to pharmacokinetic analysis using liquid chromatography coupled with tandem mass spectrometry (Cayman Chemical, Ann Arbor, MI). Tumor and pancreatic tissues were harvested, powdered on liquid nitrogen in Covaris bags, and stored at -80°C. For tissue sample processing, approximately 30 mg of tumor or pancreatic tissue was lysed on ice in mPER lysis buffer (Thermo Fisher Scientific, Waltham, MA) supplemented with 3X Halt protease inhibitor, 3X Halt phosphatase inhibitor, 3X Sigma phosphatase inhibitor cocktail 2, and 3X EDTA, and then homogenized using a Bead Ruptor 96 (Omni, Kennesaw, GA). Samples were then centrifuged at 21,000g for 10 minutes at 4°C. The clarified lysates were then transferred to ice-cold conical-bottom 96-well plates, sealed, and then stored at -80°C. The next day, samples were thawed on ice and then centrifuged at 3739g for 10 minutes at 4°C. The clarified lysates were then transferred to ice-cold round-bottom 96-well plates.Protein concentration was determined using a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA). Lysates were normalized to 10 μg / μl before boiling in 4X SDS-sample buffer and reducing agent. Samples were separated using SDS-PAGE and then analyzed using Western blot to quantify phospho-GCN2 (Thr899) (Abcam, Cambridge, UK), total GCN2, ATF4, and beta-actin (Cell Signaling Technology, Danvers, MA). Membranes are imaged with a LI-COR Odyssey CLx Imaging System (LI-COR, Lincoln, NE). Data is analyzed using GraphPad Prism software (GraphPad, San Diego, CA) to calculate % inhibition.

[0239] Compound 2, in combination with ASNase, inhibits GCN2-mediated ATF4 levels in a CCRF-CEM leukemia xenograft model. As shown in Figure 7, Compound 2 inhibited ATF4 levels by 87-91% compared to vehicle control at 50 mg / kg oral dose and by 57-93% compared to vehicle control at 25 mg / kg oral dose. Corresponding plasma levels of Compound 2 were determined at PD time points 2, 6, and 10 hours post-dose. Example 42. MV-4-11 Xenograft Efficacy Model

[0240] The MV-4-11 xenograft model was performed in an AAALAC-accredited facility in accordance with all laws, regulations, and guidelines of the National Institutes of Health (NIH), as well as with approval from the Animal Care and Use Committee of Crown Bioscience (Taicang, China). Food and water were provided ad libitum. All mice were observed for clinical signs at least once daily. Female NOD / SCID mice (6-9 weeks old, Vital River Laboratories Research Models and Services, Beijing, China) were inoculated subcutaneously in the right flank with 5 million cells in Dulbecco's phosphate-buffered saline:Matrigel (1:1) using a 27-gauge needle and syringe. Tumor volumes averaged 150 mm on day 7. 3 When the tumor volume reached 100 mg / kg / day, mice were randomly assigned to groups such that the mean tumor volume for all groups was within 10% of the overall mean tumor volume for the study population. Groups were treated on days 7-21 as follows: vehicle control (oral and IP administration to mimic the combination group) (n=10); Compound 2 orally administered at 50 mg / kg / day of Compound 2 (n=10); Leunase (Kyowa Kirin, Japan) IP administered at 1000 U / kg / day (n=10); Leunase (Kyowa Kirin, Japan) IP administered at 1000 U / kg / day (n=10) and Compound 2 orally administered at 50 mg / kg / day of Compound 2 (n=10). Tumor volumes and body weights were measured three times weekly. Tumor volume (mg) was calculated using the formula: tumor volume (mg=mm 3 )=(length x width 2 ) / 2.

[0241] Compound 2 inhibits MV-4-11 tumor growth when administered orally in combination with ASNase. As shown in Figure 8, no significant effect on tumor growth was observed after 14 days of administration of 50 mg / kg / day of Compound 2 as a single agent. Compound 2 at 50 mg / kg / day in combination with Leunase resulted in a mean tumor growth inhibition of 54% after 14 days of administration. Example 43. Recombinant GCN2 activation assay

[0242] Recombinant GCN2 (EIF2AK4) (Carna Biosciences, Japan) was dephosphorylated by incubation with 16,000 units of lambda phosphatase (New England Biolabs, Ipswich, MA) in 1X phosphatase reaction buffer at 30°C for 3 hours. To measure compound 2-mediated GCN2 regulation, 12.5 nM of dephosphorylated GCN2 diluted in kinase buffer (Invitrogen, Carlsbad, CA) was incubated with compound 2 titer at room temperature for 30 minutes. After incubation, 130 nM of substrate (GFP-eIF2a) (ThermoFisher Scientific, Waltham, MA) and 0.05 mM ATP were added to the mixture and incubated at room temperature for 60 minutes. The reaction was stopped by the addition of 10 mM EDTA, and phosphorylated eiF2a substrate was detected using 2 nM LanthaScreen Tb-anti-peIF2a(pSer52) antibody (ThermoFisher Scientific, Waltham, MA) followed by incubation in the dark. After 60 min of incubation at room temperature, TR-FRET was monitored using an excitation wavelength of 340 nm and emission wavelengths of 490 nm and 520 nm.

[0243] In Figure 9A, compound 2 unexpectedly activated the GCN2 enzyme up to 250% of control at concentrations ranging from 0.05 to 3 nM. At concentrations above 10 nM, compound 2 inhibited the activity of recombinant GCN2 enzyme in vitro. Example 44. Recombinant PERK activation assay

[0244] To measure compound 2-mediated PERK regulation, 200 nM of PERK enzyme (Enzo, Farmingdale, NY) was incubated with compound 2 titer for 30 minutes at room temperature. After incubation, 200 nM of substrate (GFP-eIF2a) (ThermoFisher Scientific, Waltham, MA) and 1 mM ATP were added to the mixture and incubated at room temperature for 60 minutes. The reaction was stopped by adding 10 mM EDTA and phosphorylated eiF2a substrate was detected using 2 nM of LanthaScreen Tb-anti-peIF2a (pSer52) antibody (ThermoFisher Scientific, Waltham, MA), followed by incubation in the dark. After 60 minutes of incubation at room temperature, TR-FRET was monitored using an excitation wavelength of 340 nm and emission wavelengths of 490 nm and 520 nm.

[0245] In Figure 9B, concentrations of Compound 2 ranging from 0.05 to 6 nM unexpectedly activated the PERK enzyme up to 250% of controls. At concentrations above 3 nM, Compound 2 inhibited the activity of recombinant PERK enzyme in vitro. Example 45. Spheroid Western Blot Assay

[0246] NSCLC cell lines H358 (catalog no. CRL-5807) and H2030 (catalog no. CRL-5914), and fibrosarcoma cell line HT-1080 (catalog no. CRL-5807) were obtained from American Type Culture Collect (ATTC, Manassas, VA). Cells were grown in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen, Carlsbad, CA) and 1% penicillin / streptomycin / L-glutamine at 37°C, 5% CO2, and 95% humidity and maintained at 50-80% confluence until use. Cells were seeded at 500,000 cells / well in 24-well Elplasia plates (Corning, Glendale, AZ) containing 1 mL / well of complete growth medium and incubated overnight at 37°C, 5% CO2, and 95% humidity to form spheroids. The next day, serial dilutions of test compounds were prepared in complete growth medium, and 500 μL of existing medium was removed from each well and replaced with fresh medium containing compound titer. Spheroids were then incubated with test compounds under the same conditions for 4 or 24 hours. At the end of incubation, spheroids were washed with PBS supplemented with 1× Halt protease inhibitor, 1× Halt phosphatase inhibitor (Invitrogen, Carlsbad, CA), 1× Sigma phosphatase inhibitor cocktail 2 (Sigma, St. Louis, MO) and 1× EDTA (Invitrogen, Carlsbad, CA), and then lysed with M-PER Mammalian Protein Extraction Reagent (Invitrogen, Carlsbad, CA) supplemented with 3× inhibitor mix as described above. Cell lysates were sonicated in a water bath sonicator (Qsonica, Newtown, CT), and the supernatant was boiled with SDS buffer and reducing agent. Western blots were performed to quantify phospho-GCN2 (R&D Systems, Minneapolis, MN), ATF4, CHOP, cleaved PARP, cleaved caspases 3 and 7, and beta-actin (Cell Signaling Technology, Danvers, MA).Membranes were imaged with a LI-COR Odyssey CLx Imaging System (LI-COR, Lincoln, NE).

[0247] In Figures 10A, 10B, and 10C, compound 2 unexpectedly demonstrated stimulation of the ISR and apoptotic pathways in solid tumor cancer cell lines (Figure 10A, H2030; Figure 10B, H358; Figure 10C, HT-1080). Western blot images of upregulated GCN2 phosphorylation (pGCN2), as well as downstream signaling proteins ATF4 and pro-apoptotic pathway markers (cleaved-PARP1, cleaved-Caspase 3 / 7) (actin as loading control) are shown in Figures 10A, 10B, and 10C. Levels of pGCN2 were upregulated in H2030 and H358 cell lines, while ATF4, c-PARP, and c-Caspase 7 were upregulated over a concentration range from 4.6 nM up to 300 nM. Western blot quantification of ATF4 signal is represented in bar graph format. Compound 2 upregulated ATF4 by up to 20-fold in H2030 spheroids, 12-fold in H358 spheroids, and 15-fold in HT-1080 spheroids. Example 46. Spheroid proliferation assay

[0248] Non-small cell lung cancer (NSCLC) cell lines H358 (catalog no. CRL-5807) and H2030 (catalog no. CRL-5914), and fibrosarcoma line HT-1080 (catalog no. CRL-5807) were obtained from American Type Culture Collect (ATTC, Manassas, VA). Cells were grown in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 1% penicillin / streptomycin / L-glutamine (Invitrogen, Carlsbad, CA) at 37°C, 5% CO2, and 95% humidity and maintained at 50–80% confluency until use. 500-1000 cells were seeded into 96-well ultra-low attachment plates (Corning, Glendale, NC) with complete growth medium at 80 μL / well and incubated overnight at 37 °C, 5% CO2, and 95% humidity to allow spheroids to form. The next day, serial dilutions of test compounds were prepared in complete growth medium and 20 μL / well medium containing compound titers was added to the spheroids in triplicate. Spheroids were then incubated with test compounds under the same conditions for 5 days. At the end of the incubation, spheroids were lysed with 100 μL / well of CellTiter-Glo 3D Viability Assay (Promega, Madison, WI) and luminescence was detected using a Synergy2 or equivalent reader (Biotek, Winooski, VT). Data were analyzed using GraphPad Prism software (GraphPad, San Diego, CA) to determine IC 50 The value was calculated.

[0249] In Figures 11A, 11B, and 11C, Compound 2 as a single agent induced 50% cell regression in H2030 solid tumor spheroids (Figure 11A). Compound 2 as a single agent induced 50% cell regression in H358 (IC 50 10 nM, Fig. 11B) and HT-1080 (IC 50 12 nM, FIG. 11C) inhibited solid tumor spheroids.

[0250] In Figures 12A, 12B, and 12C, Compound 2 showed additive or synergistic effects on the inhibition of spheroid growth when combined with standard of care (SOC) agents. Representative graphs are shown in Figures 12A, 12B, and 12C. Figure 12A represents a cell proliferation experiment of H2030 NSCLC spheroids treated with Compound 2 titers in combination with 3.3 nM sotorasib (diamonds) or without sotorasib (circles) for 96 hours. Figure 12B represents a cell proliferation experiment of H358 NSCLC spheroids treated with Compound 2 titers in combination with 3.3 nM sotorasib (diamonds) or without sotorasib (circles) for 96 hours. Figure 12C depicts cell proliferation experiments of H2030 NSCLC spheroids treated with Compound 2 titers in combination with 3.7 nM trametinib (triangles) or without trametinib (circles) for 96 hours. In all three experiments, combination treatment resulted in greater spheroid regression compared to either single agent. Example 47. HT-1080 shRNA knockdown assay

[0251] Fibrosarcoma cell line HT-1080 (catalog no. CRL-5807) was obtained from American Type Culture Collect (ATCC, Manassas, VA). Cells were grown in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum and 1% penicillin / streptomycin / L-glutamine (Invitrogen, Carlsbad, CA) at 37°C, 5% CO2, and 95% humidity and maintained at 50-80% confluence until use. Constructs expressing shRNA targeting PERK, GCN2, or a scrambled control were obtained from MISSION (Sigma, St. Louis, MO), and lentiviruses containing each construct were produced in-house. Cells were seeded at 400,000 cells / well in 6-well tissue culture plates (ThermoFisher Scientific, Waltham, MA) with complete growth medium and incubated overnight at 37°C, 5% CO2, and 95% humidity to allow attachment. The next day, existing medium was removed from each well and replenished with 1 mL / well of fresh medium containing 5 μg / mL polybrene (VectorBuilder, Boston, IL). Cells were transduced with 200 μL / well of the corresponding lentivirus for 24 h and then processed for Western blot and cell proliferation assays.

[0252] In Figure 13A, total GCN2 was knocked down by 57% using targeted shRNA in HT-1080 cell line. PERK was knocked down by 65% ​​using targeted shRNA in HT-1080 cell line. Compound 2-mediated upregulation of ISR markers (ATF4, pEIF2a, and CHOP) was downregulated by knockdown of GCN2 or PERK. Figure 13B shows Western blot quantification of ATF4 signal. Compound 2-mediated upregulation of ATF4 was reduced by 75% by knockdown of GCN2. Compound 2-mediated upregulation of ATF4 was reduced by 50% by knockdown of PERK. In Figure 13C, in this solid tumor cell line, compound 2-mediated spheroid growth inhibition was attenuated by knockdown of GCN2, but not PERK. Example 48. HT-1080 fibrosarcoma xenograft pharmacokinetic / pharmacodynamic (PK / PD) model

[0253] The HT-1080 xenograft model was performed in an AAALAC-accredited facility in accordance with all laws, regulations, and guidelines of the National Institutes of Health (NIH) and with approval from the Animal Care and Use Committee at Labcorp (Ann Arbor, MI). Food and water were provided ad libitum. All mice were observed for clinical signs at least once daily. Female nude mice (6-7 weeks old) were inoculated subcutaneously just below the right upper axilla with 2 million cells in Dulbecco's phosphate-buffered saline using a 27-gauge needle and syringe. Tumor volume averaged 150 mm on day 6. 3When the tumor reached 10%, mice were randomly assigned to groups such that the mean tumor burden for all groups was within 10% of the overall mean tumor burden for the study population. Groups were treated on days 6-11 as follows: vehicle control (oral dosing) (n=9); Compound 2 orally administered at 10 mg / kg BID (n=9); Compound 2 orally administered at 5 mg / kg BID; Compound 2 orally administered at 1 mg / kg BID 2, 6, and 10 hours after dosing on day 11, prior to sample collection (n=9). Blood samples were collected in K2EDTA tubes, processed to plasma, flash frozen in liquid nitrogen, and then stored at -80°C. Plasma samples were subjected to pharmacokinetic analysis using liquid chromatography coupled with tandem mass spectrometry (Cayman Chemical, Ann Arbor, MI). Tumors were harvested, powdered over liquid nitrogen in Covaris bags, and stored at -80°C. For tissue sample processing, approximately 30 mg of tumor tissue was lysed on ice in mPER lysis buffer (Thermo Fisher Scientific, Waltham, MA) supplemented with 3X Halt protease inhibitor, 3X Halt phosphatase inhibitor, 3X Sigma phosphatase inhibitor cocktail 2, and 3X EDTA, then homogenized using a Bead Ruptor 96 (Omni, Kennesaw, GA). Samples were then centrifuged at 21,000g for 10 minutes at 4°C. The clarified lysates were then transferred to ice-cold conical-bottom 96-well plates, sealed, and then stored at -80°C. The next day, samples were thawed on ice and then centrifuged at 3739g for 10 minutes at 4°C. The clarified lysates were then transferred to ice-cold round-bottom 96-well plates. Protein concentrations were determined using a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA). Lysates were normalized to 10 μg / μl before boiling in 4× SDS-sample buffer and reducing agent. Samples were resolved using SDS-PAGE and then analyzed using Western blot to quantify ATF4 and beta-actin (Cell Signaling Technology, Danvers, MA).Membranes were imaged with a LI-COR Odyssey CLx Imaging System (LI-COR, Lincoln, NE). Data were analyzed and % inhibition was calculated using GraphPad Prism software (GraphPad, San Diego, CA).

[0254] In Figures 14 and 14B, compound 2 upregulated ATF4 levels in HT-1080 fibrosarcoma PK / PD xenograft model. As shown in Figure 14A, compound 2 upregulated ATF4 levels 4.5-fold compared to vehicle control when orally administered at 10 mg / kg BID, upregulated ATF4 levels 3.5-fold compared to vehicle control when orally administered at 5 mg / kg BID, and upregulated ATF4 levels 3-fold compared to vehicle control when orally administered at 1 mg / kg BID. The corresponding plasma levels of compound 2 were determined at PD time points 2, 6, and 10 hours after administration (Figure 14B). Example 49. LoVo xenograft efficacy model

[0255] The LoVo xenograft model was performed in an AAALAC-accredited facility in accordance with all laws, regulations, and guidelines of the National Institutes of Health (NIH) and with approval from the Animal Care and Use Committee of Crown Bioscience (Taicang, China). Food and water were provided ad libitum. All mice were observed for clinical signs at least once daily. Female Balb / c nude mice (5-9 weeks old) were inoculated subcutaneously just below the right upper axilla with 10 million cells in Dulbecco's phosphate-buffered saline using a 27-gauge needle and syringe. Tumor volume averaged 100 mm on day 6. 3 When tumor mass reached 100 mg / kg, mice were randomly assigned to groups such that the mean tumor burden for all groups was within 10% of the overall mean tumor burden for the study population. Groups were treated on days 6-27 as follows: vehicle control (oral administration) (n=9), Compound 2 orally administered at 5 mg / kg BID (n=9), Compound 2 orally administered at 1 mg / kg BID. Tumor volumes and body weights were measured three times weekly. Tumor mass (mg) was calculated using the formula: tumor mass (mg = mm 3 )=(length x width2 ) / 2. Example 50. HT-1080 xenograft efficacy model

[0256] The HT-1080 xenograft model was performed in an AAALAC-accredited facility in accordance with all laws, regulations, and guidelines of the National Institutes of Health (NIH) and with approval from the Animal Care and Use Committee at Labcorp (Ann Arbor, MI). Food and water were provided ad libitum. All mice were observed for clinical signs at least once daily. Female nude mice (6-7 weeks old) were inoculated subcutaneously just below the right upper axilla with 2 million cells in Dulbecco's phosphate-buffered saline using a 27-gauge needle and syringe. Tumor burden averaged 100 mm on day 6. 3 When tumor mass reached 100 mg / kg, mice were randomly assigned to groups such that the mean tumor burden for all groups was within 10% of the overall mean tumor burden for the study population. Groups were treated on days 6-14 as follows: vehicle control (oral administration) (n=9), Compound 2 orally administered at 10 mg / kg BID (n=9), Compound 2 orally administered at 5 mg / kg BID. Tumor volumes and body weights were measured three times weekly. Tumor mass (mg) was calculated using the formula: tumor mass (mg = mm 3 )=(length x width 2 ) / 2.

[0257] As shown in Figure 15A, compound 2 inhibited tumor growth in a LoVo colorectal xenograft model. The mean tumor growth inhibition was 69% after Compound 2 was administered at 5 mg / kg BID for 22 days. In Figure 15B, compound 2 inhibited tumor growth in a HT-1080 fibrosarcoma xenograft model. The mean tumor growth inhibition was 50% after Compound 2 was administered at 10 mg / kg BID for 14 days.

[0258] Equivalent Although specific embodiments have been discussed, the above specification is illustrative and not limiting. Many variations of the embodiments will become apparent to those skilled in the art upon review of this specification. The full scope of what is disclosed should be determined by reference to the claims, along with their full scope of equivalents, and this specification, along with such variations.

[0259] Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and the like used in the specification and claims are to be understood as being modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained.

Claims

1. Compounds of formula IA: 【Chemistry 1】 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein: X １ and X ３ are each independently selected from the group consisting of CH and N; X ２ is NR ６ , O, and S; R １ , R ２ and R ３ are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R ４ is selected from the group consisting of halogen, alkoxy, and alkyl; R ５ is selected from the group consisting of H, halogen and alkyl; R ６ is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R ７ is selected from the group consisting of H, alkyl, and acyl, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof.

2. (i) at least one of R 1 , R 2 and R 3 is halogen; and / or (ii) at least one of R 1 , R 2 and R 3 is fluoro; and / or (iii) R 1 is fluoro; and / or (iv) The compound of claim 1, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein X 1 is N.

3. (i) A compound represented by formula I-B: 【Chemistry 2】 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein: X 1 and X 3 are each independently selected from the group consisting of CH and N; X 2 is selected from the group consisting of NR 6 , O, and S; R 2 and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy, and alkyl; R 5 is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acyl, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof; or (ii) A compound represented by formula IC: 【Transformation 3】 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein: X 2 is selected from the group consisting of NR 6 , O, and S; X 3 is selected from the group consisting of CH and N; R 2 and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy, and alkyl; R 5 is selected from the group consisting of H, halogen, and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; R 7 is selected from the group consisting of H, alkyl, and acyl, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof; or (iii) A compound represented by formula ID: 【Chemistry 4】 or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein: R 2 and R 3 are each independently selected from the group consisting of H, halogen, cyano, and alkoxy; R 4 is selected from the group consisting of halogen, alkoxy, and alkyl; R 5 is selected from the group consisting of H, halogen and alkyl; R 6 is selected from the group consisting of H, alkyl, alkenyl, alkenylalkyl, alkynyl, alkynylalkyl, cycloalkyl, cycloalkylalkyl, alkoxyalkyl, cycloalkenyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, and heteroarylalkyl; 10. The compound of claim 1, wherein R7 is selected from the group consisting of H, alkyl, and acyl, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof.

4. (i) R 2 is H and R 3 is H; or (ii) R 2 is F and R 3 is H; or (iii) The compound of claim 3, wherein R 2 is H and R 3 is F, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof.

5. R 6 is (i) (C 1 -C 8 ) alkyl, (C 2 -C 8 ) alkenyl, (C 2 -C 8 ) alkenyl-(C 1 -C 4 ) alkyl, (C 2 -C 8 ) alkynyl, (C 2 -C 8 ) alkynyl-(C 1 -C 4 ) alkyl, (C 3 -C 8 ) cycloalkyl, (C 3 -C 8 ) cycloalkyl-(C 1 -C 4 ) alkyl, (C 3 -C 8 ) alkoxy-(C 1 -C 4 ) alkyl, (C 3 -C 8 ) cycloalkenyl, (C 3 -C 8 ) cycloalkenyl-(C 1 -C 4 ) alkyl, heterocyclyl, heterocyclyl-(C 1 -C 4 ) selected from the group consisting of alkyl, aryl, heteroaryl, and heteroaryl-(C 1 -C 4 )alkyl; or (ii) selected from the group consisting of (C 1 -C 8 )alkyl, (C 3 -C 8 )cycloalkyl, (C 3 -C 8 )alkoxy-(C 1 -C 4 )alkyl, heterocyclyl, and heteroaryl; or (iii) A compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, selected from the group consisting of: 【Transformation 5】

6. R 4 is (i) selected from the group consisting of halogen, (C 1 -C 6 )alkoxy, and (C 1 -C 6 )alkyl; or (ii) The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, selected from the group consisting of chloro, fluoro, methoxy, and methyl.

7. R 5 is (i) selected from the group consisting of H, halogen, and (C 1 -C 6 ) alkyl; or (ii) The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, selected from the group consisting of chloro, fluoro, and methyl.

8. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, wherein R 7 is H.

9. A compound selected from the group consisting of: 【Transformation 6】 and pharmaceutically acceptable salts, enantiomers, stereoisomers, and tautomers thereof.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 4 and 9, or a pharmaceutically acceptable salt, enantiomer, stereoisomer, or tautomer thereof, and a pharmaceutically acceptable carrier or excipient.

11. A pharmaceutical composition according to claim 10 for use in treating a disorder.

12. The pharmaceutical composition of claim 10 for use in a patient in need thereof, said use comprising the pharmaceutical composition: (i) a method for treating a disease caused by dysregulation of the integrated stress response and / or the unfolded protein response; or (ii) A method for treating a disease caused by dysregulation of the integrated stress response and / or the unfolded protein response, wherein the dysregulation of the integrated stress response and / or the unfolded protein response is caused by GCN2 kinase (general control nonderepressible 2 kinase) or PERK kinase (Protein kinase R-like Endoplasmic Reticulum kinase); or (iii) a method for regulating the activity of GCN2 kinase (general control nonderepressible 2 kinase); or (iv) A method for inhibiting the activity of GCN2 kinase (general control nonderepressible 2 kinase); or (v) a method for activating the activity of GCN2 kinase (general control nonderepressible 2 kinase); or (vi) A method for regulating the activity of PERK kinase (Protein kinase R-like Endoplasmic Reticulum kinase); or (vii) A method for inhibiting the activity of PERK kinase (Protein kinase R-like Endoplasmic Reticulum kinase); or (viii) A method for activating the activity of PERK kinase (Protein kinase R-like Endoplasmic Reticulum kinase); or (ix) A method for inhibiting GCN2 kinase (general control nonderepressible 2 kinase) and PERK kinase (Protein kinase R-like Endoplasmic Reticulum kinase); or (x) a method for treating cancer; or (xi) a method for treating amyloidosis; or (xii) a method for treating light chain amyloidosis; or (xiii) A method for treating a disease selected from a GCN2-related disease and a PERK-related disease; or (xiv) A method for treating a disease selected from a GCN2-related disease and a PERK-related disease, the method further comprising administering to the patient a therapeutically effective amount of one or more therapeutic agents, wherein the one or more therapeutic agents are: (1) is selected from the group consisting of an IMiD agent, a proteasome inhibitor, a steroid, an anti-CD38 agent, an anti-CD20 agent, a Bcl-2 inhibitor, a PI3K inhibitor, a bispecific antibody, a nucleoside analog, a BTK inhibitor, a DNA alkylating agent, an EZH2 inhibitor, an anthracycline, a topoisomerase inhibitor, a platin, a tyrosine kinase inhibitor, an HDAC inhibitor, a nuclear export inhibitor, a microtubule inhibitor, L-asparaginase, a pegylated asparaginase, a PERK inhibitor, an mTOR inhibitor, an immunomodulator, a MAPK pathway inhibitor, a MEK inhibitor, an ERK inhibitor, and a Ras inhibitor; or (2) L-asparaginase, pegaspargase, calaspargase pegol-mnkl, bortezomib, carfilzomib, ixazomib, thalidomide, pomalidomide, lenalidomide, dexamethasone, prednisone, daratumumab, daratumumab / hyaluronidase, isatuximab, rituximab, obinutuzumab, venetoclax, idelalisib, copanlisib, duvelisib, umbralisib, gemcitabine, cytarabine , ibrutinib, acalabrutinib, zanubrutinib, bendamustine, cyclophosphamide, tazemetostat, doxorubicin, daunorubicin, etoposide, oxaloplatin, carboplatin, cisplatin, bosutinib, dasatinib, imatinib, nilotinib, ponatinib, panobinostat, selinexor, vincristine, JZP-458, eriaspase, PF745 (JZP-341), asparaginase Erwinia chrysanthemi (crisantaspase), Escherichia coli asparaginase (colaspase), anti-PD1 agents, anti-PDL1 agents, and anti-CTLA4 agents.

13. The use is a method for treating cancer, and the cancer is selected from the group consisting of colorectal cancer, lung cancer, mesothelioma, pancreatic cancer, pharyngeal cancer, laryngeal cancer, esophageal cancer, stomach cancer, duodenal cancer, small intestine cancer, breast cancer, ovarian cancer, testicular tumor, prostate cancer, liver cancer, thyroid cancer, kidney cancer, uterine cancer, gestational choriocarcinoma, brain tumor, retinoblastoma, skin cancer, melanoma, sarcoma, fibrosarcoma, malignant bone tumor, bladder cancer, blood cancer, leukemia, and acute myeloid leukemia.

13. The pharmaceutical composition for use according to claim 12, wherein the cancer is selected from the group consisting of acute lymphoblastic leukemia, chronic lymphoblastic leukemia, chronic myelogenous leukemia, multiple myeloma, B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, T-cell lymphoma, erythroleukemia, histiocytic lymphoma, Waldenstrom's macroglobulinemia, light chain amyloidosis, and malignant lymphoma.

14. The cancer is (i) leukemia; or (ii) acute myeloid leukemia; or (iii) acute lymphoblastic leukemia; or (iv) fibrosarcoma; or (v) multiple myeloma; or (vi) lymphoma; or (vii) B-cell lymphoma; or (viii) T-cell lymphoma. A pharmaceutical composition for the use according to claim 13.

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