N-(3-(benzo[b]thiophene-2-carboxamido)-phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide derivatives and related compounds as lactate / ATP production inhibitors for the treatment of cancer

Abstract

The present invention relates to a compound of formula (I) or an enantiomer, diastereomer, N-oxide, solvate or pharma- ceutically acceptable salt thereof. The present invention further relates to a compound of formula (I) for use in a method for the prophylaxis and / or treatment of a disease or condition, in particular cancer. JPEG2025511974000226.jpg37106

Description

[Technical field]

[0001] The present invention relates to novel inventive compounds, particularly compounds useful in the treatment of cancer. The present invention further relates to pharmaceutical compositions comprising such compounds. The present invention further relates to such compounds for use as medicaments. The present invention further relates to such compounds or such pharmaceutical compositions for use in a method for preventing and / or treating a disease or condition in a particular cancer. The present invention further relates to a method for treating a disease or condition mediated by the lactate / ATP mechanism in a particular cancer, wherein an effective amount of such compounds or such pharmaceutical compositions is administered to a subject in need thereof. The present invention further relates to the use of such compounds or such pharmaceutical compositions in the manufacture of a medicament for treating a disease or condition mediated by the lactate / ATP mechanism in a particular cancer. [Background technology]

[0002] Cancer remains the second leading cause of mortality in mankind. Fatal cancers are those that are diagnosed too late, i.e., at a very late stage of disease progression, or those that have been treated but not cured. After repeated recurrences or at late stages, i.e., typically stage 3B or higher, many solid cancers tend to acquire phenotypes that are particularly resistant to chemo- and radiotherapy. Such advanced and treatment-resistant cancers are often characterized by a high degree of metastasis, where the primary tumor and / or metastases adopt a high degree of the so-called Warburg metabolism (Weber et al., Front Oncol. (2016) 6:257).

[0003] Nearly 100 years ago, Otto Warburg, as this process was later named, discovered that cancer cells utilize anaerobic glycolysis to convert glucose to lactate. This metabolic phenotype produces ATP with low efficiency compared to oxidative phosphorylation, i.e., mitochondrial respiration. Warburg ascribed this metabolic switch from respiration toward anaerobic glycolysis as a result of a lack of oxygen in the core of advanced cancers (Warburg et al., Biochem Zeitschr. (1924) 152: 309-44; Warburg et al., J Gen Physiol (1927) 8 (6): 519-30). However, research over the past two decades has revealed that this metabolic switch toward the dominance of anaerobic glycolysis occurs to meet the enormous demand for carbon-containing building blocks for malignant cell growth, rather than to achieve efficient ATP production. Several intermediates of glycolysis or the pentose phosphate pathway are used by cells to synthesize nucleosides, lipids, different carbohydrates or amino acids, or to generate NADPH, which is required for either lipid or cholesterol biosynthesis or to regenerate defense systems that protect against reactive oxygen species (ROS) (Vander et al., Cell (2017) 168 (4): 657-669). ROS are dangerous for the cells mentioned above, and their production rate increases dramatically in cancer cells (Hosios et al., J Biol Chem. (2018) 293 (20): 7490-7498). Therefore, pathways such as the glutathione or thioredoxin family that detoxify ROS species are absolutely required in rapidly growing cells and highly demand the provision of NADPH-reducing equivalents, which are typically supplied by the first oxidative step of the pentose phosphate pathway, which uses glucose as a substrate (Wang et al., Theranostics. (2011) 11(10):4839-4857).

[0004] ROS stress can also cause damaged proteins, misfolded or chemically damaged proteins to induce the so-called proteotoxic stress or unfolded protein response (UPR). When this stress situation occurs in the endoplasmic reticulum (ER), it is recognized as ER stress, a subphenomenon of proteotoxic stress (Radanovic et al., Cells. (2021) 10(11):2965). As part of the proteotoxic stress response, certain transcription factors, in particular heat shock factor-1 (HSF-1), are well known as master regulators of the transcriptional program that occurs to counteract and resolve proteotoxic stress. HSF-1 is one of the first proteins induced under heat shock or chemically induced proteotoxic stress responses and coordinates protein transcription and therefore production as well as systems that deal with this specific stress situation, such as certain chaperones, ROS-detoxification systems, certain membrane transporters or pumps (Dai et al., Philos Trans R Soc Lond B Biol Sci. (2018) 373(1738):20160525).

[0005] All of the above elements, increased anaerobic glycolysis, increased flux through the pentose phosphate pathway, increased need for ROS protection and dealing with proteotoxic stress, may be defined as hallmarks of Warburg metabolism in a broader and more modern definition.

[0006] The final major result of Warburg metabolism is the production of high levels of lactate, which is an inevitable consequence of not utilizing oxidative phosphorylation. Lactate can be considered as a waste product of anaerobic glycolysis resulting from the action of lactate dehydrogenase (LDH), which reduces pyruvate to lactate with the concomitant oxidation of NADH to NAD+. This NAD+ needs to be regenerated to allow intermediate conversions along the way of glycolysis, catalyzed by GAPDH. Thus, the conversion of pyruvate to lactate closes the loop of NAD-regeneration in anaerobic glycolysis, not allowing pyruvate to enter mitochondria for oxidative phosphorylation metabolism. However, importantly, cancer cells do not completely shut down mitochondrial metabolism when deprived of oxygen, using Warburg metabolism predominantly. This is essential as mitochondrial metabolism, especially the citric acid cycle, allows the use of alternative Warburg fuels, namely amino acids such as asparagine or glutamine, to provide other amino acids and pyruvate using this central metabolic turntable. These acids can be partially converted to lactate for the reasons mentioned above, or can satisfy the demand for carbon building blocks via "reverse" glycolysis, i.e., gluconeogenesis and the pentose phosphate pathway (Baltazar et al., Front Oncol. (2020) 10:231).

[0007] Lactate is therefore a common end product of various aspects of the complex Warburg metabolic state. Moreover, lactate has an additional function by suppressing immune responses within the lactate-producing tissue environment. The main physiological consequence of the immunosuppressive role of lactate and the associated local acidic pH is that skeletal muscles produce large amounts of lactate, especially during exercise and when oxygen supply is insufficient. In untrained muscles, the microfibers tend to break, exposing intracellular components to the extracellular environment. This is a typical stimulus for cell-resident antigen-presenting cells and other myeloid cells to define damaged muscle as a target for the immune system. To prevent this, both lactate and the acidic pH act as immunosuppressants themselves, thereby disabling unwanted and unnecessary immune attacks on muscles.

[0008] This physiological immunosuppressive role of lactate is used by solid tumors which release significant amounts of lactate in the tumor microenvironment, thereby preventing immune destruction of the tumor. In this respect, such tumor compartments resemble muscle "sores," and several publications have demonstrated immunosuppressive activity in essentially all relevant immune cell subtypes.

[0009] In summary, lactate is primarily a waste product of Warburg metabolism, and its determination in the context of cancer cell proliferation is probably the best indicator of the Warburg process, since it integrates carbon flux from different sources under anaerobic and aerobic conditions.

[0010] Second, lactate plays a major role as an immunosuppressant in the tumor microenvironment, so reducing tumor lactate production is an attractive goal to achieve in clinical cancer therapy (Vaupel et al., J Physiol. (2021) 599(6):1745-1757; de la Cruz-Lopez et al., Oncol. (2019) 9:1143).

[0011] However, it must be considered that cancer cells and transiently active skeletal muscle cells are not the only cell types in the human body that adopt Warburg metabolism. It is well documented that all cells undergoing several proliferation cycles under a certain proliferation stimulus adopt this metabolic phenotype, since it is an evolutionarily predetermined pattern that supports cell growth and division. In particular, T cells that are sensitized to antigens and receive strong proliferation signals also consume large amounts of glucose and convert it into lactate. Nevertheless, it must be considered that T cells, whose activity is very beneficial in the immune antitumor response, typically proliferate in compartments in close contact with antigen-presenting cells, i.e., tumor-draining lymph nodes, etc. Although it can be clearly demonstrated that T cell proliferation upon CD3 / CD28 stimulation can be completely blunted by glycolytic inhibitors, such as 2-deoxyglucose or others, it may be possible to separate the antiglycolytic effect on tumor cells from the antiglycolytic effect on expanding T cells by targeting the respective pharmaceutical agents to such tumors and excluding lymph nodes or compartments. Once T cells have expanded to become CD8-positive antitumor effector cells, they enter the tumor and subsequently attack cancer cells. This antitumor efficacy is not necessarily blunted by lowering lactate in the tumor, as illustrated by a recent publication (Renner et al., Cell Rep. (2019) 29(1):135-150.e9).

[0012] So far, there are no drugs available that can inhibit lactate production in the context of cancer treatment, generally without cytotoxic side effects. There have been several attempts, using a variety of different chemical compounds, either to directly inhibit one or more of the glycolytic enzymes, or to downregulate or inhibit the main transcription factors or master regulators of the Warburg program. The most prominent examples of the latter mode of action, or members of the family of hypoxia-inducible factors (HIF), are, foremost, Hif-1 alpha or Hif-2 alpha. Hif-1, -2 or -3 alpha heterodimerize with a common partner, Hif-1 beta, also known as ARNT (aryl hydrocarbon receptor nuclear transporter). All two members of the Hif-family are well described to act as transcription factors inducing key genes mediating the Warburg effect, namely GLUT-1 (glucose transporter), HK2, PK-M, LDH-A, and MCT-4, the latter being a hypoxic and tumor-specific isoform of glycolytic enzymes or lactate transporter (Yu et al., J Cancer. (2017) 8(17):3430-3440).

[0013] Most of these drugs failed during clinical development either due to lack of efficacy in terms of anti-cancer activity or because they exhibited severe and intolerable side effects.

[0014] The first non-transformed cell type sensitive to glycolysis inhibition is the erythrocyte, which relies solely on glycolysis as its energy source. As a result, direct glycolysis inhibition leads to hemolysis, which manifests clinically as hemolytic anemia. Moreover, cancer patients under chemotherapy frequently suffer from anemia, making this side effect even more unacceptable in the context of cancer treatment (Oshima et al., Cell Rep. (2020) 30 (6): 1798-1810. e4).

[0015] Another approach to limit cancer cell proliferation targets the proteotoxic stress response. This can be accomplished in a variety of ways, including but not limited to, inhibitors of the proteasome, such as bortezomib or carfilzomib. Limiting proteasomal protein degradation leads to the accumulation of misfolded proteins and ultimately to apoptosis.

[0016] There have been attempts to inhibit HSF-1 as a key regulator of the proteotoxic stress response, to reduce the extent to which cells are affected and to alleviate the anti-proliferative effects of proteotoxic stress. One such chemical compound that has been found to downregulate HSF-1 is CCT251236, a small molecule chemical compound derived from screening for the above effects. CCT251236 has been published to limit cell proliferation in several different cancer cell lines. This compound also inhibited tumor growth in an ovarian cancer xenograft mouse model. CCT251236, and its closely related PROTAC analog CCT367766, were found to bind strongly to Pirin. However, Pirin is still not proven to be a functionally relevant target of these bisamide compounds, based on published results showing that CCT251236 still exhibits micromolar inhibitory activity in other related proteins, such as acetylcholinesterase or certain kinases. Nevertheless, CCT251236 has potent antitumor efficacy at a dose of 30 mg / kg delivered orally every other day (Cheeseman et al., J Med Chem. (2017) 60(1):180-201; Chessum et al., J Med Chem. (2018) 61(3):918-933).

[0017] WO 2015 / 049535 and WO 2016 / 156872 disclose a series of bisamides overlapping with certain compounds named CCT245232 and CCT251236, which are described in more detail in Cheeseman et al.; J Med Chem (2017), 60(1) 180-201 and in Chessum et al. J Med Chem. (2018); 61(3): 918-933. CCT245232 was identified in a screen for inhibitors of the HSF-1 proteotoxic stress pathway. Using medicinal chemistry efforts, CCT245232 was further developed into CCT251236, which demonstrated better solubility and oral bioavailability in mice. Compounds in this bisamide series demonstrated potent antiproliferative activity against cancer cells, with SK-OV-3 as the reference cell line, and demonstrated precise growth IC 50 CCT251236 also exhibits significant anti-cancer activity in mouse xenograft models of cancer. [ka]

[0018] The anticancer activity of this bisamide-type compound was attributed to its inhibitory effect on the HSF-1 proteotoxic stress pathway.

[0019] Chessum et al.,J Med Chem.(2018)61(3):918-933) disclose that these compounds bind very strongly to Pirin, a non-heme iron containing cytoplasmic protein, with cellular functions that have only been described so far. In Meyers et al.(ACS Med Chem Lett(2018);9(12):1199-1204.), these bisamide compounds are demonstrated to bind indiscriminately to different kinases as protein targets. Therefore, it is not entirely clear whether these bisamide compounds actually exert their anticancer effects solely by binding to Pirin.

[0020] Autophagy is a highly conserved catabolic mechanism that degrades cellular components to provide intermediate metabolites during cellular stress, e.g., starvation (J Pathol. (2010) 221: 3-12). Although autophagy can act to overcome temporary nutrient depletion in tumor cells, uncontrolled or inappropriate autophagy can induce ferroptosis, a type of regulated cell death caused by iron-dependent lipid peroxidation (Cell Research (2016) 26 1021-1032). Ferroptosis is dependent on autophagy and is primarily mediated by the release of iron bound to intracellular proteins, resulting in biotoxicity.

[0021] Autophagy is considered to be the main mechanism impaired in certain diseases, especially age- and lifestyle-related diseases (Madeo et al., Cell Metab. 5; 29(3): 592-610 (2019). Restoring autophagy capacity in the respective organs and cells may overcome the accumulation of small fibrillar protein tangles of beta-amyloid, alpha-nuclein and other proteins, which are involved in the pathogenesis of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), but also in age-related memory decline in general. Beyond the application in neurodegenerative diseases, activation of autophagy may also have beneficial effects in fibrotic lung diseases, obesity and diabetes, as well as in viral infections, where induction of autophagy may be helpful (Carmona-Gutierrez et al., Microb Cell, 7(5): 119-128. (2020).

[0022] Small molecule cancer therapy has so far been largely limited to cytotoxic drugs, i.e., agents that target biological enzymes, structural proteins such as microtubules, or DNA, in order to prevent cancerous growth at the expense of dramatic side effects on normal healthy proliferating tissues, particularly intestinal epithelium or hair follicles.

[0023] The so-called biguanides, exemplified by the drugs metformin, buformin and phenformin, are a class of drugs known to lead to side effects resulting from stimulation of glycolysis. These three drugs, phenformin being the most potent and metformin the least potent, are or were antidiabetic drugs that improve insulin sensitivity, thereby ameliorating hyperglycemia, a major hallmark of type 2 diabetes. All biguanides are said to result in a decrease in mitochondrial ATP production by inhibiting complex I of the mitochondrial respiratory chain (Bridges et al., BMC Biol. 14, 65 (2016)), which favors ATP production through upregulation of glycolysis. This explains the main problem of biguanides in clinical use: the possible occurrence of lactic acidosis. The incidence of lactic acidosis, which has a mortality rate of approximately 50%, is 64 events in 100,000 patients treated with phenformin and 3 events in 100,000 patients treated with metformin. Phenformin is clearly a more powerful drug and was widely used as an antidiabetic drug in the 1960s and 70s, but has been supplanted by the weaker but safer metformin. Several studies investigating the incidence of different cancers in type 2 diabetes patients with or without metformin treatment have shown that metformin reduced the incidence of cancer in these patients, as well as the overall mortality rate. However, clinical trials have shown a small benefit of metformin treatment in cancer patients, but there are no data from prospective clinical studies comparing standard chemotherapy with or without additional metformin treatment in terms of survival or cancer response, although data from preclinical and restospective studies suggest this (Top et al., Pharmaceuticals (Basel). 15(3): 312 (2022)).

[0024] The main limitation of the use of metformin as an adjunct to existing chemotherapy is its modest effectiveness against cancer at doses used in antidiabetic treatment. For the treatment of cancer, further increases in metformin dosage, or alternatively, even more intensive use of phenformin, may be indicated, but the known side effect of lactic acidosis with phenformin and the fact that phenformin has been taken off the market prevent this.

[0025] Therefore, in addition to new cancer treatments, there is also a need for co-treatments that reduce lactate production that would otherwise limit the dose of biguanide drugs that can be given to each patient. The present subject matter addresses both of these needs. Summary of the Invention

[0026] Unexpectedly, in addition to reducing the production of intracellular ATP and extracellular lactate, reducing intracellular HIF1α, and inducing endoplasmic reticulum stress genes, in particular CHAC and CHOP, the compounds of the invention have been found to activate autophagy in cells treated with the compounds.

[0027] In experimental results, the compounds of the present invention also show broad antiproliferative activity against several cancer cell lines in vitro.The compounds of the present invention have been successfully used to limit tumor growth in certain mouse models of cancerous proliferation.

[0028] The compounds of the invention also demonstrate the potential to strongly reduce lactate production, allowing them to be suitably co-administered with biguanide drugs to achieve otherwise unattainable biguanide levels in vivo, thus allowing tumors to be effectively treated while reducing or eliminating the risk of inducing biguanide dose-limiting lactic acidosis.

[0029] The compounds of the present invention target a previously unexplored mechanism, which is the induction of proteotoxic stress in the endoplasmic reticulum, which is exemplified by well-known markers of this response, such as Chac1 or Chop. As a result, the cell responds by upregulating genes whose function may help to overcome the proteotoxic stress, for example by producing more glutathione. System Xc - SLC7A11, also known as SLC7A11, is a cysteine-glutamate antiporter that exhibits upregulation of such a response.

[0030] The induction of autophagy exerted by the compounds of the present invention tends to be downstream of the main proteotoxic stress response, and is a reflection of the overall shutdown of most metabolic pathways by the compounds of the present invention.The sequence of events established by using these compounds is that they first induce the proteotoxic stress response, which results in a global quiescence, i.e., the shutdown of cells in terms of metabolic activity that results in less lactate production.This simulates a kind of cell starvation, and induces autophagy as a countermeasure to resupply cells with more metabolic fuel.

[0031] The significant reduction in lactate production can not only be used to directly target the proliferation of cancer cells that exhibit the hallmarks of Warburg metabolism, i.e. high glycolytic rates and high lactate production, but this drastic but non-cytotoxic shutdown of cell proliferation and metabolism can also be exploited to counter the side effects of other drugs that can simulate glycolysis and in turn dangerously increase lactate concentrations in the blood, resulting in lactic acidosis as a well-known side effect.

[0032] In a first aspect, the present invention relates to a compound according to formula (I) or an enantiomer, diastereomer, N-oxide, solvate or pharma- ceutically acceptable salt thereof, [ka] In the formula, A, D, K, L, X, Y, Z, R 1 -R 4 and the indices n, p, q and r are defined for said compounds as disclosed below.

[0033] In a further aspect, the present invention relates to a pharmaceutical composition comprising a compound according to formula (I) as defined above and a pharma- ceutically acceptable excipient.

[0034] In a further aspect, the present invention relates to a compound according to formula (I) for use as a medicament.

[0035] In a further aspect, the present invention relates to a compound according to formula (I) as defined above or to a pharmaceutical composition comprising a compound according to formula (I) as defined above for use in a method for the prevention and / or treatment of a disease or condition mediated by the lactate / ATP mechanism.

[0036] In a further aspect, the present invention relates to a method for preventing and / or treating a disease or condition mediated by the lactate / ATP mechanism, said method comprising administering to a subject in need thereof an effective amount of said compound according to formula (I) or said pharmaceutical composition comprising said compound according to formula (I).

[0037] In a further aspect, the present invention also relates to the use of a compound according to formula (I) in the preparation of a medicament for the prevention and / or treatment of a disease or condition mediated by the lactate / ATP mechanism.

[0038] In a further aspect, the present invention relates to a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), for use in a method for the prevention and / or treatment of a disease or condition, preferably cancer, by inducing endoplasmic reticulum stress in cells affected by said disease or condition, thereby reducing intracellular ATP and lactate production in said cells.

[0039] In a further aspect, the present invention relates to a method for preventing and / or treating a disease or condition, preferably cancer, comprising administering to a subject a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), wherein said prevention and / or treatment is achieved by inducing endoplasmic reticulum stress in cells affected by said disease or condition, thereby reducing intracellular ATP and lactate production in said cells.

[0040] In a further aspect, the present invention relates to the use of a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), in the manufacture of a medicament for the prevention and / or treatment of a disease or condition, preferably cancer, by inducing endoplasmic reticulum stress in cells affected by said disease or condition, thereby reducing intracellular ATP and lactate production in said cells.

[0041] In a further aspect, the present invention relates to a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) for use in a method for preventing and / or treating cancer (preferably cancer cells and / or cancerous tumors), wherein said compound reduces Hif1-alpha at protein levels in hypoxic areas of said cancer, thereby downregulating ATP and lactate production in these areas and ultimately leading to a reduction (preferably an efficient reduction) of the growth of hypoxic areas and of said cancer.

[0042] In a further aspect, the present invention relates to a method for preventing and / or treating cancer (preferably cancer cells and / or cancerous tumors), comprising administering to a subject a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), wherein said prevention and / or treatment is achieved by reducing Hif1-alpha at the protein level in hypoxic areas of said cancer, thereby downregulating ATP and lactate production in these areas and ultimately leading to a reduction (preferably an efficient reduction) of the growth of hypoxic areas and of said cancer.

[0043] In a further aspect, the present invention relates to the use of a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), in the manufacture of a medicament for preventing and / or treating cancer (preferably cancer cells and / or cancerous tumors) by reducing Hif1-alpha at protein levels in hypoxic areas of said cancer, thereby downregulating ATP and lactate production in these areas and ultimately leading to a reduction (preferably an efficient reduction) of the growth of hypoxic areas and of said cancer.

[0044] In a further aspect, the present invention relates to a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) for use in a method for the prevention and / or treatment of a disease or condition susceptible to the induction of endoplasmic reticulum stress, wherein administration of said compound or said pharmaceutical composition to a subject having or suspected of having said disease or condition results in a reduction in the protein levels of Hif-1 alpha.

[0045] In a further aspect, the present invention relates to a method for preventing and / or treating a disease or condition susceptible to the induction of endoplasmic reticulum stress, comprising administering to a subject a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), wherein administration of said compound to a subject having or suspected of having said disease or condition results in a reduction in the protein levels of Hif-1 alpha.

[0046] In a further aspect, the present invention relates to the use of a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), in the manufacture of a medicament for the prevention and / or treatment of a disease or condition susceptible to the induction of endoplasmic reticulum stress by reducing the protein level of Hif-1 alpha.

[0047] In a further aspect, the present invention relates to a compound according to formula (I), or a pharmaceutical composition comprising said compound according to formula (I), for use in a method for the prevention and / or treatment of an autophagy-related disease selected from neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and Huntington's disease, amyotrophic lateral sclerosis, metabolic disorders such as obesity and type I and II diabetes mellitus, viral infections and diseases leading to the accumulation of certain debris particles, such as fibrotic lung diseases.

[0048] In a further aspect, the present invention relates to a method for preventing and / or treating an autophagy-related disease selected from neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and Huntington's disease, amyotrophic lateral sclerosis, metabolic disorders such as obesity and type I and II diabetes, viral infections and diseases leading to the accumulation of certain debris particles, such as fibrotic lung diseases, said method comprising administering to a subject a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I).

[0049] In a further aspect, the present invention relates to the use of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) in the manufacture of a medicament for the prevention and / or treatment of an autophagy-related disease selected from neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and Huntington's disease, amyotrophic lateral sclerosis, metabolic disorders such as obesity and type I and II diabetes mellitus, viral infections and diseases leading to the accumulation of certain debris particles, such as fibrotic lung diseases, wherein said method comprises administering to a subject a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I).

[0050] In a further aspect, the present invention relates to the use of a compound according to formula (I) in the preparation of a medicament for the prevention and / or treatment of cancer.

[0051] In a further aspect, the invention relates to a composition, preferably a pharmaceutical composition, comprising a compound according to formula (I) and a biguanide, and to said composition for use as a medicament.

[0052] In a further aspect, the invention relates to a combination of a compound according to formula (I) and a biguanide for use in therapy.

[0053] In a further aspect, the present invention relates to a method for preventing and / or treating a disease comprising administering to a subject a combination of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide.

[0054] In a further aspect, the present invention relates to a combination of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide for use in a method for the prevention and / or treatment of cancer.

[0055] In a further aspect, the present invention relates to a method for preventing and / or treating cancer, said method comprising administering to a subject a combination of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide.

[0056] In a further aspect, the present invention relates to the use of a combination of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide in the manufacture of a medicament for the prevention and / or treatment of cancer.

[0057] As used herein, administering a compound according to formula (I) or a pharmaceutical composition comprising a compound according to formula (I) to a subject should be understood as administering an effective amount of said compound or said pharmaceutical composition to said subject.

[0058] In a further embodiment of the above treatment and use, said compound according to formula (I) is comprised in a pharmaceutical composition together with a pharma- ceutical acceptable excipient. [Brief description of the drawings]

[0059] [Figure 1A] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1B] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1C] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1D] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1E] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1F] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1G]FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1H] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1I] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1J] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1K] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 1L] FIG. 1 shows downregulated ATP levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2A] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2B] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2C] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2D] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2E] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2F] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2G] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2H] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2I] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2J]FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2K] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 2L] FIG. 2 shows that extracellular lactate levels were downregulated in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cell cultures after 48 hours of treatment with the compounds of Examples 3 / 16, 3 / 8 and 10. [Figure 3A] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3B] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3C] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3D] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3E] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3F] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3G]FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3H] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3I] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3J] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3K] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 3L] FIG. 3 shows ER stress genes induced in HeLa cells over a 4 hour period of treatment with 10 nM of Examples 3 / 16, 3 / 8 and 10 compounds. [Figure 4A] FIG. 4 shows that compounds of Examples 3 / 8 and 10 down-regulate HIF1α protein levels in HeLa cells after 24 hours of treatment under normoxia and hypoxia. [Figure 4B] FIG. 4 shows that compounds of Examples 3 / 8 and 10 down-regulate HIF1α protein levels in HeLa cells after 24 hours of treatment under normoxia and hypoxia. [Figure 5A] Figure 5 shows that the compounds of Examples 3 / 8, 3 / 16 and 10 activate autophagy in HeLa cells, and autophagy is prevented by application of bafilomycin A. Similarly, Examples 3 / 8, 3 / 16 and 10 induce phosphorylation of p70-S6K (Thr389) in HeLa cells and in cells treated simultaneously with bafilomycin A. [Figure 5B]Figure 5 shows that the compounds of Examples 3 / 8, 3 / 16 and 10 activate autophagy in HeLa cells, and autophagy is prevented by application of bafilomycin A. Similarly, Examples 3 / 8, 3 / 16 and 10 induce phosphorylation of p70-S6K (Thr389) in HeLa cells and in cells treated simultaneously with bafilomycin A. [Figure 5C] Figure 5 shows that the compounds of Examples 3 / 8, 3 / 16 and 10 activate autophagy in HeLa cells, and autophagy is prevented by application of bafilomycin A. Similarly, Examples 3 / 8, 3 / 16 and 10 induce phosphorylation of p70-S6K (Thr389) in HeLa cells and in cells treated simultaneously with bafilomycin A. [Figure 6A] FIG. 6 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 directly induce autophagy in a HeLa reporter cell line expressing eGFP-LC3 (ns, not significant; ****, p<0.0001). [Figure 6B] FIG. 6 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 directly induce autophagy in a HeLa reporter cell line expressing eGFP-LC3 (ns, not significant; ****, p<0.0001). [Figure 6C] FIG. 6 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 directly induce autophagy in a HeLa reporter cell line expressing eGFP-LC3 (ns, not significant; ****, p<0.0001). [Figure 6D] FIG. 6 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 directly induce autophagy in a HeLa reporter cell line expressing eGFP-LC3 (ns, not significant; ****, p<0.0001). [Figure 7A] FIG. 7 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 reduce the formation of lactate produced by HeLa cells in a dose-dependent manner when treated with phenformin. [Figure 7B]FIG. 7 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 reduce the formation of lactate produced by HeLa cells in a dose-dependent manner when treated with phenformin. [Figure 7C] FIG. 7 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 reduce the formation of lactate produced by HeLa cells in a dose-dependent manner when treated with phenformin. [Figure 7D] FIG. 7 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 reduce the formation of lactate produced by HeLa cells in a dose-dependent manner when treated with phenformin. [Figure 7E] FIG. 7 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 reduce the formation of lactate produced by HeLa cells in a dose-dependent manner when treated with phenformin. [Figure 7F] FIG. 7 shows that the compounds of Examples 3 / 16, 3 / 8 and 10 reduce the formation of lactate produced by HeLa cells in a dose-dependent manner when treated with phenformin. [Figure 8A] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8B] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8C] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8D] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8E] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8F] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8G] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8H] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8I] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8J]FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8K] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8L] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8M] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8N] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8O] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8P]FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8Q] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8R] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8S] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8T] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8U] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8V]FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8W] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8X] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8Y] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8Z] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8AA] FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. [Figure 8AB]FIG. 8 shows a comparison of the activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in downregulating ATP (FIGS. 8A and 8B), extracellular lactate levels (FIGS. 8C and 8D) and induction of ER stress genes (FIGS. 8E-8AB) in Panc02 and SKOV3 cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0060] The present invention relates to a compound of formula (I) or an enantiomer, diastereomer, N-oxide, solvate or pharma-ceutically acceptable salt thereof, [ka] During the ceremony K is O, S or C(R 5 ) 2 and; L is O, S or C(R 5 ) 2 and; n is 0, 1, 2, 3 or 4; p is 1 or 2; q are, independently of each other, 0, 1 or 2; r is 1 or 2; A is C 1-6 Alkyl, halogen, CN, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, OH, -OC 1-6 Alkyl or O-Halo-C 1-6 is alkyl; X is S, O, or NR 6 , N or CR 7 and; Y is S, O, NR 6 , N or CR 7 and; Z is C or N; D represents a 6-membered aryl or a 5-6-membered heteroaryl containing 1-4 heteroatoms independently selected from O, N, and S; R1 , F, C 1-3 Alkyl or spirocyclic fused C 3-6 is cycloalkyl; R 2 are independently halogen or C 1-6 is alkyl; R 3 is hydrogen, halogen, C 1-6 Alkyl, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, -OC 1-6 Alkyl or O-Halo-C 1-6 is alkyl; R 4 is hydrogen, halogen or V 1 -V 2 -V 3 -V 4 -V 5 and Where: V 1 is absent, a bond, or C 1-6 is alkylene; V 2 is absent, a bond, or O, S, S(O) x , S(O)(=NR a ), S(O) 2 NR a , N.R. a , N.R. a C(O) or C(O)NR a and; V 3 is absent, a bond, or C 1-6 is alkylene; Here, V 1 Or V 3 C in 1-6 The alkylene is unsubstituted or is selected from the group consisting of OH, C 1-6 Alkyl, C 1-6 Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen, and oxo; or V 1 Or V 3 C in1-6 Two C's on alkylene 1-6 Alkyl groups, together with the carbon atom to which they are attached, form a C 3-6 forming a cycloalkyl group; V 4 is absent, a bond, or O, S, S(O) x , S(O)(=NR a ), S(O) 2 NR a , N.R. a , N.R. a C(O) or C(O)NR a and; V 5 are hydrogen, halogens, OH, CN, CO 2 H, CO 2 -C 1-6 Alkyl, N(R a ) 2 , C 1-6 Alkyl, C 3-8 cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, phenyl, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl, phenyl or heteroaryl is unsubstituted or is selected from the group consisting of CN, CO 2 H, CO 2 -C 1-6 Alkyl, N(R a ) 2 , O.H., C. 1-6 Alkyl, OC 1-6 Alkyl, C(O)-C 1-6 Alkyl, C 1-6 Haloalkyl, halogen, oxo, spiro fused rings C 3-6 cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; V5 has been replaced by; R 5 is hydrogen, F, C1-3 Alkyl or spirocyclic fused C 3-6 is cycloalkyl; R 6 is hydrogen or C 1-6 is alkyl; R 7 is hydrogen, halogen or C 1-6 is alkyl; R a is hydrogen or C 1-3 is alkyl; X is 1 or 2.

[0061] In an embodiment in combination with any of the above or below embodiments, the compound is represented by formula (I), or an enantiomer, diastereomer, N-oxide, solvate, or pharma- ceutically acceptable salt thereof: Formula (I) [ka] During the ceremony K is O, S or C(R 5 ) 2 and; L is O, S or C(R 5 ) 2 and; n is 0, 1, 2, 3 or 4; p is 1 or 2; q are, independently of each other, 0, 1 or 2; r is 1 or 2; A is C 1-6 Alkyl, halogen, CN, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, OH, -OC 1-6 Alkyl or O-Halo-C 1-6 is alkyl; X is S, O, or NR 6 , N or CR 7 and; Y is S, O, NR 6 , N or CR 7 and; Z is C or N; D represents a 6-membered aryl or a 5-6-membered heteroaryl containing 1-4 heteroatoms independently selected from O, N, and S; R 1 , F, C 1-3 Alkyl or spirocyclic fused C 3-6 is cycloalkyl; R 2 are independently halogen or C 1-6 is alkyl; R 3 is hydrogen, halogen, C 1-6 Alkyl, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, -OC 1-6 Alkyl or O-Halo-C 1-6 is alkyl; R 4 is hydrogen, halogen or V 1 -V 2 -V 3 -V 4 -V 5 and where V 1 is absent, a bond, or C 1-6 is alkylene; V 2 is absent, a bond, or O, S, S(O) x , S(O)(=NR a ), S(O) 2 NR a , N.R. a , N.R. a C(O) or C(O)NR a and; V 3 is absent, a bond, or C 1-6 is alkylene; Here, V 1 Or V 3 C in 1-6 The alkylene is unsubstituted or 1-6 Alkyl, C 1-6Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen, and oxo; or V 1 Or V 3 C in 1-6 Two C's on alkylene 1-6 Alkyl groups, together with the carbon atom to which they are attached, form a C 3-6 forming a cycloalkyl group; V 4 is absent, a bond, or O, S, S(O) x , S(O)(=NR a ), S(O) 2 NR a , N.R. a , N.R. a C(O) or C(O)NR a and; V 5 are hydrogen, halogens, OH, CN, CO 2 H, CO 2 -C 1-6 Alkyl, N(R a ) 2 , C 1-6 Alkyl, C 3-8 cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, phenyl, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl, phenyl or heteroaryl is unsubstituted or is selected from the group consisting of CN, CO 2 H, CO 2 -C 1-6 Alkyl, N(R a ) 2 , O.H., C. 1-6 Alkyl, OC 1-6 Alkyl, C(O)-C 1-6 Alkyl, C 1-6 Haloalkyl, halogen, oxo, spiro fused rings C 3-6cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; V5 has been replaced by; R 5 is hydrogen, F, C 1-3 Alkyl or spirocyclic fused C 3-6 is cycloalkyl; R 6 is hydrogen or C 1-6 is alkyl; R 7 is hydrogen, halogen or C 1-6 is alkyl; R a is hydrogen or C 1-3 is alkyl; X is 1 or 2.

[0062] In an embodiment in combination with any of the above or below embodiments, the compound is represented by formula (II): Formula (II) [ka] U 1 , U 2 , U 3 and U 4 is independently selected from the group consisting of N and C.

[0063] In an embodiment in combination with any of the above or below embodiments, the compound is represented by formula (III): Formula (III) [ka] During the ceremony, U 1 and U 4 is independently selected from N and C; X is S, O, or NR 6 or N; Y is CR7 Or N.

[0064] In any embodiment in combination with any of the above or below embodiments, the index n is 0.

[0065] In any embodiment in combination with the above or below embodiments, the index p is 1.

[0066] In an embodiment in combination with any of the above or below embodiments, the compound is represented by formula (IV): Formula (IV) [ka] During the ceremony A is C 1-6 is alkyl or halogen; X is S or NH.

[0067] In any combination with the above or below embodiments, R 4 is hydrogen, halogen or V 1 -V 2 -V 3 -V 4 -V 5 where: V 1 is absent, a bond, or C 1-4 is alkylene; V 2 is absent, a bond, or O, S, S(O) x , N.R. a and; V 3 is absent, a bond, or C 1-4 is alkylene, V 4 is absent, a bond, or O, S, S(O) x , N.R. a and Here, V 1 Or V 3C in 1-4 Alkylene is unsubstituted or 1-3 Alkyl, C 1-3 Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen, and oxo; or V 1 Or V 3 C in 1-4 Two C's on alkylene 1-6 Alkyl groups, together with the carbon atom to which they are attached, form a C 3-6 forming a cycloalkyl group; V 5 are hydrogen, halogens, OH, CN, C 1-6 Alkyl, C 3-6 cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl or heteroaryl is unsubstituted or is selected from the group consisting of OH, C 1-6 Alkyl, OC 1-6 Alkyl, C(O)-C 1-6 Alkyl, C 163 Haloalkyl, halogen, oxo, spiro fused rings C 3-6 cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; V5 has been replaced by

[0068] In any combination with the above or below embodiments, R 4 is hydrogen, halogen or V 2 -V 3 -V 5 where: V 2 is absent, a bond or O; V 3 is absent, a bond, or C 1-4 is alkylene, Here, V 3 C in 1-4 Alkylene is unsubstituted or 1-3 substituted with 1 to 4 substituents independently selected from the group consisting of alkyl; V 5 are hydrogen, halogens, OH, CN, C 1-6 alkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl or heteroaryl is unsubstituted or is selected from the group consisting of OH, C 1-6 Alkyl, OC 1-6 Alkyl, C 1-6 1 to 4 substituents R independently selected from the group consisting of haloalkyl, halogen and oxo; V5 has been replaced by

[0069] In an embodiment in combination with any of the above or below embodiments, the compound is selected from the following group: and an N-oxide, solvate or pharma-ceutically acceptable salt thereof: [ka] [ka] [ka] [ka] [ka] [ka]

[0070] In formula (I), the variables K and L are independently O, S, or C(R 5 ) 2 In a preferred embodiment in combination with any of the above or below embodiments, one of the variables K and L represents O and the other represents S, or both variables represent O, or both variables represent S, more preferably both K and L represent O. In a further preferred embodiment in combination with any of the above or below embodiments, one of K and L represents O and the other represents C(R 5 )

[0071] In formula (I), the variable A is C 1-6 Alkyl, halogen, CN, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, OH, -OC 1-6 Alkyl or O-Halo-C 1-6 In any combination with the above or below embodiments, the variable A is C 1-6 Alkyl, halogen, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, OH, -OC 1-6 Alkyl or O-Halo-C 1-6 In any combination with the above or below embodiments, the variable A is C 1-6 Alkyl, halogen, halo-C 1-6 Alkyl, OH, -OC 1-6 Alkyl or O-Halo-C 1-6 In a preferred embodiment in combination with any of the above or below embodiments, the variable A is C 1-6It is alkyl or halogen.

[0072] In a more preferred embodiment in combination with any of the above or below embodiments, the variable moiety A is 3 , F or Cl. In a most preferred embodiment in combination with any of the above or below embodiments, the variable A is F.

[0073] In formula (I), the index q is independently selected from 0, 1 and 2. In a preferred embodiment in combination with any of the above or below embodiments, the index q is independently selected from 0 or 1. In a more preferred embodiment in combination with any of the above or below embodiments, the index q is 0.

[0074] In formula (I), the index p is 1 or 2. In a preferred embodiment in combination with any of the above or below embodiments, the index p is 1.

[0075] In formula (I), the index n is 0, 1, 2, 3 or 4. In a preferred embodiment in combination with any of the above or below embodiments, said index n is 0, 1 or 2. In a more preferred embodiment in combination with any of the above or below embodiments, said index n is 0 or 1. In a most preferred embodiment in combination with any of the above or below embodiments, said index n is 0.

[0076] In any embodiment in combination with any of the above or below embodiments, the variable A is F and the index q is 0.

[0077] In any of the above or below embodiments in combination, the variables K and L both represent O, n is 0, p is 1 and q is 0.

[0078] In formula (I), the following structural moiety [ka] Ring D in the formula (I) represents a 6-membered aryl or a 5- to 6-membered heteroaryl containing 1 to 4 heteroatoms independently selected from O, N and S.

[0079] In a preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0080] Each of the above groups representing ring D is R 4 and 1 or 2 R 3 has been replaced by

[0081] In a preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0082] Each of the above groups representing ring D is R 4 and 1 or 2 R 3 has been replaced by

[0083] In a further embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0084] Each of the above groups representing ring D is R 4 and 1 or 2 R 3 has been replaced by

[0085] In any combination with the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0086] Each of the above groups representing ring D is R 4 and 1 or 2 R 3 has been replaced by

[0087] In a preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0088] Each of the above groups representing ring D is R 4and 1 or 2 R 3 has been replaced by

[0089] In a more preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] but: [ka]

[0090] The phenyl ring representing ring D is R 4 and 1 or 2 R 3 has been replaced by

[0091] In an equally preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] but: [ka]

[0092] The pyridine ring representing the ring D is R 4 and 1 or 2 R 3 has been replaced by

[0093] In any combination with the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0094] In a preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0095] In a further embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0096] In any combination with the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0097] In a preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] is selected from the group consisting of: [ka]

[0098] In a more preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] But the following is true: [ka]

[0099] In an equally preferred embodiment in combination with any of the above or below embodiments, the structural unit [ka] But the following is true: [ka]

[0100] In formula (I), R 1 , F, C 1-3 Alkyl or spirocyclic fused C 3-6 It is cycloalkyl.

[0101] In a preferred embodiment in combination with any of the above or below embodiments, R 1 is F or C 1-3 It is an alkyl.

[0102] In formula (I), R 2 is halogen or C1-6 It is an alkyl.

[0103] In a preferred embodiment in combination with any of the above or below embodiments, R 2 is F, Cl or C 1-3 It is an alkyl.

[0104] In formula (I), R 3 is hydrogen, halogen, C 1-6 Alkyl, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl, halo-C 3-6 Cycloalkyl, -OC 1-6 Alkyl or O-Halo-C 1-6 It is an alkyl.

[0105] In a preferred embodiment in combination with any of the above or below embodiments, R 3 are hydrogen, F, Cl, Br, C 1-6 Alkyl, halo-C 1-6 Alkyl, C 3-6 Cycloalkyl or -OC 1-6 alkyl, more preferably R 3 is F, Cl or C 1-6 It is an alkyl.

[0106] In formula (I), R 4 is hydrogen, halogen or V 1 -V 2 -V 3 -V 4 -V 5 where: V 1 is absent, a bond, or C 1-6 is alkylene; V 2 is absent, a bond, or O, S, S(O) x , S(O)(=NR a ), S(O) 2 NR a , N.R. a , N.R. a C(O) or C(O)NR aand; V 3 is absent, a bond, or C 1-6 is alkylene; Here, V 1 Or V 3 C in 1-6 The alkylene is unsubstituted or is selected from the group consisting of OH, C 1-6 Alkyl, C 1-6 Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen, and oxo; or V 1 Or V 3 C in 1-6 Two C's on alkylene 1-6 Alkyl groups, together with the carbon atom to which they are attached, form a C 3-6 forming a cycloalkyl group; V 4 is absent, a bond, or O, S, S(O) x , S(O)(=NR a ), S(O) 2 NR a , N.R. a , N.R. a C(O) or C(O)NR a and; V 5 are hydrogen, halogens, OH, CN, CO 2 H, CO 2 -C 1-6 Alkyl, N(R a ) 2 , C 1-6 Alkyl, C 3-8 cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, phenyl, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl, phenyl or heteroaryl is unsubstituted or is selected from the group consisting of CN, CO 2 H, CO 2 -C 1-6Alkyl, N(R a ) 2 , O.H., C. 1-6 Alkyl, OC 1-6 Alkyl, C(O)-C 1-6 Alkyl, C 1-6 Haloalkyl, halogen, oxo, spiro fused rings C 3-6 cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; V5 has been replaced by

[0107] In a preferred embodiment in combination with any of the above or below embodiments, R 4 is hydrogen, halogen or V 1 -V 2 -V 3 -V 4 -V 5 where: V 1 is absent, a bond, or C 1-4 is alkylene; V 2 is absent, a bond, or O, S, S(O) x , N.R. a and; V 3 is absent, a bond, or C 1-4 is alkylene, V 4 is absent, a bond, or O, S, S(O) x , N.R. a and Here, V 1 Or V 3 C in 1-4 Alkylene is unsubstituted or is selected from the group consisting of OH, C 1-3 Alkyl, C 1-3 Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen, and oxo; or V 1 Or V 3C in 1-4 Two C's on alkylene 1-6 Alkyl groups, together with the carbon atom to which they are attached, form a C 3-6 forming a cycloalkyl group; V 5 are hydrogen, halogens, OH, CN, C 1-6 Alkyl, C 3-6 cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl or heteroaryl is unsubstituted or is selected from the group consisting of OH, C 1-6 Alkyl, OC 1-6 Alkyl, C(O)-C 1-6 Alkyl, C 1-6 Haloalkyl, halogen, oxo, spiro fused rings C 3-6 cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; V5 has been replaced by

[0108] In a preferred embodiment in combination with any of the above or below embodiments, R 4 -H, -CH 3 , -CF 3 , -OCH 3 , F, Cl, Br, [ka] [ka] It is.

[0109] In a more preferred embodiment in combination with any of the above or below embodiments, R 4 teeth, [ka] It is.

[0110] In yet another more preferred embodiment in combination with any of the above or below embodiments, R 4 teeth, [ka] It is.

[0111] In a preferred embodiment in combination with any of the above or below embodiments, the compound is selected from the group below, and an N-oxide, solvate or pharma-ceutically acceptable salt thereof: [ka]

[0112] In an equally preferred embodiment in combination with any of the above or below embodiments, the compound is selected from the group consisting of: and N-oxides, solvates or pharma-ceutically acceptable salts thereof: [ka] [ka]

[0113] The compounds of the present invention according to formula (I) exhibit surprising biological activity.

[0114] Lactate is at the crossroads between tumor metabolism and the late-stage cancer microenvironment, surviving the body's natural immune response. Specifically, lactate is the end product of the so-called Warburg metabolism, a metabolism commonly found in cancer cells that favors fermentative over oxidative pathways. Warburg metabolism converts glucose to lactate, diverting carbon flux into energy to support the growth of ever-dividing cancer cells. At the same time, lactate and the associated acidic pH in the tumor environment also exert an immunosuppressive effect that reduces the ability of immune cells to fight against new or existing cancer cells. This "lactate immune shield" tolerates only those immune cells that are themselves passive against the tumor using Warburg metabolism. These so-called myeloid-derived suppressor cells (MDSCs) support cancer by signaling other active immune cells to avoid the tumor, thereby protecting the growing tumor by helping it escape immune attack. And overall, increased lactate and associated acidity in the tumor microenvironment promotes tumor growth, both directly, by driving tumor growth itself, and indirectly, by inhibiting the body's ability to fight back against tumor growth.

[0115] It has been found that the compounds of the present invention inhibit lactate production. This activity reverses the above-mentioned tumor-promoting process. Specifically, without being bound by theory, reducing lactate concentration in cancer cells that depend on lactate as an energy source has the effect of starving cancer cells of the energy they need to grow, thereby slowing or even stopping cancer growth. At the same time, reducing lactate concentration in the tumor microenvironment weakens the "lactate immune shield" that would otherwise normally protect cancer cells, thereby favorably exposing existing cancer cells to attack by cells of the body's own immune system. Thus, tumors are hindered in their ability to produce new cells, making them more vulnerable to immune attack and destruction.

[0116] Specifically, through investigation, it was revealed that the compounds of the present invention not only inhibit cancerous proliferation, but that this inhibition is mediated by an unprecedented reduction in ATP and lactate in cancer cell cultures (Figures 1, 2). A unique feature of this ATP-decreasing mechanism is that it occurs as a biphasic effect. In the first phase, at concentrations ranging from single nanomolar to low single micromolar depending on the actual compound of the present invention, the heteroaryl bisamide compounds of the present invention reduce ATP and lactate and inhibit cell proliferation, but do not induce cell death, i.e., apoptosis or necrosis. Therefore, the first phase can be called "cytostatic". Further analysis revealed that the compounds of the present invention induce markers of endoplasmic reticulum stress, such as CHAC-1, Chop, XBP-1, or SLC7A11, in the same concentration range in which they exhibited cytostatic effects (Figure 3, H. Ren et al, Front. Aging Neurosci. (2021), 13: 691881). The induction of these ER stress markers, as determined by quantitative real-time PCR, preceded a concentration-dependent reduction in Hif1-alpha protein levels as determined from the same cancer cells (Figure 4). Since Hif1-alpha plays a major role as a master regulator of the transcriptional control of Warburg effect-related genes under hypoxic conditions in tumors, this effect is likely one mechanism for the strong antiproliferative effect observed for the compounds of the present invention.

[0117] Thus, the applicant not only provides novel heteroaryl-core containing bisamide compounds with unprecedented activity against a variety of cancer cell lines and no predicted genotoxicity, but also identifies a novel biological mechanism by which these compounds exert their anti-proliferative effects in such cancer cells.

[0118] A further aspect of the invention relates to a pharmaceutical composition comprising a compound according to any one of the above embodiments and a pharma- ceutical acceptable excipient. In a preferred embodiment, the pharmaceutical composition may comprise from about 5 wt% to about 98 wt% of one or more compounds of the invention, preferably from about 0.5 mg to about 0.5 g, and even more preferably from about 1 mg to about 100 mg of active compound. In certain preferred embodiments, the pharmaceutical composition is formulated to allow administration to a subject enterally (e.g., orally, sublingually, and / or buccally), parenterally (e.g., intravenously, intramuscularly, subcutaneously), intranasally, topically, rectally, vaginally, and / or by inhalation.

[0119] A further aspect of the invention relates to a compound according to any one of the above embodiments (i.e. a compound according to formula (I) and / or a pharmaceutical composition comprising same) for use as a medicament.

[0120] A further aspect of the invention relates to a compound according to any one of the above embodiments or a pharmaceutical composition comprising a compound according to any one of the above embodiments for use in a method for preventing and / or treating a disease or condition mediated by the lactate / ATP mechanism, in particular cancer. In certain preferred embodiments, the method comprises administering the compound or the pharmaceutical composition to a subject enterally (e.g. orally, sublingually, and / or buccally), parenterally (e.g. intravenously, intramuscularly, and / or subcutaneously), intranasally, topically, rectally, vaginally and / or by inhalation.

[0121] A further aspect of the invention relates to a method of treating a disease or condition mediated by the lactate / ATP mechanism, in certain cancers, wherein an effective amount of a compound of the invention or a pharmaceutical composition of the invention is administered to a subject in need thereof. In certain preferred embodiments, the method comprises administering the compound or pharmaceutical composition to a subject enterally (e.g., orally, sublingually, and / or buccally), parenterally (e.g., intravenously, intramuscularly, and / or subcutaneously), intranasally, topically, rectally, vaginally, and / or by inhalation.

[0122] The present invention further relates to the use of a compound of the invention or a pharmaceutical composition of the invention in the manufacture of a medicament for treating a disease or condition mediated by the lactate / ATP mechanism, in certain cancers. In certain preferred embodiments, the medicament is formulated to allow administration of the compound or pharmaceutical composition to a subject enterally (e.g., orally, sublingually, and / or buccally), parenterally (e.g., intravenously, intramuscularly, and / or subcutaneously), intranasally, topically, rectally, vaginally, and / or by inhalation.

[0123] In a preferred embodiment in combination with any of the above or below embodiments, said disease or condition mediated by the lactate / ATP mechanism is cancer.

[0124] In a preferred embodiment in combination with any of the above or below embodiments, the cancer is selected from the group consisting of cancer of the head and neck, eye, skin, mouth, throat, esophagus, breast, bone, lung, colon, sigmoid colon, rectum, stomach, prostate, uterus, breast, ovary, kidney, liver, pancreas, brain, intestine, heart, or adrenal gland. More specifically, the cancer may be a solid tumor, a sarcoma, a carcinoma, a fibrosarcoma, a myxosarcoma, a liposarcoma, a chondrosarcoma, an osteosarcoma, a chordoma, angiosarcoma, an endothelioma, a lymphangiosarcoma, a lymphangioendothelioma sarcoma, a synovial tumor, a mesothelioma, a Ewing's tumor, a leiomyosarcoma, a rhabdomyosarcoma, a colon carcinoma, a pancreatic cancer, a breast cancer, an ovarian cancer, a uterine carcinoma, adenoid cystic carcinoma, a prostate cancer, a squamous cell carcinoma, a basal cell carcinoma, an adenocarcinoma, a sweat gland carcinoma, a sebaceous gland carcinoma, a papillary carcinoma, a papillary adenocarcinoma, a cystadenocarcinoma, a medullary carcinoma, a bronchial carcinoma, a renal cell carcinoma, a liver cancer, a bile duct carcinoma, a choriomas, a seminoma, an embryonal carcinoma, a Wilms' tumor, a cervical cancer, a testicular tumor, a lung carcinoma, a small cell lung carcinoma, a non-small cell Examples of cancers that may be present include lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, blood-borne tumors, acute lymphoblastic leukemia, acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cell leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute monoblastic leukemia, acute erythroleukemic leukemia, acute megakaryoblastic leukemia, acute myelomonocytic leukemia, acute nonlymphocytic leukemia, acute anaplastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, hairy cell leukemia, or multiple myelomas (see, e.g., Encyclopedia (See, The American Cancer Society, 3rd Edition (2018) Editors Paolo Boffetta, Pierre Hainaut). In a preferred embodiment in combination with any of the above or below embodiments, the cancer is a solid tumor. According to a preferred embodiment, the cancer is selected from leukemia, melanoma, liver cancer, pancreatic cancer, lung cancer, colon cancer, brain cancer, ovarian cancer, breast cancer, prostate cancer, and kidney cancer. In another embodiment, the cancer is liver cancer, pancreatic cancer, non-small cell lung cancer, breast cancer, or kidney cancer.In a preferred embodiment in combination with any of the above or below embodiments, the cancer is preferably selected from non-small cell lung cancer, sarcoma, gynecological cancers such as cervical, ovarian cell or uterine carcinoma, adenoid cystic carcinoma, pancreatic cancer and other gastrointestinal cancers, such as gastric or esophageal cancer, when a compound of the invention or a pharmaceutical composition of the invention is administered in combination with a biguanide.

[0125] A type of cancer may be characterized, without limitation, as hypoxic and / or hyperglycolytic, as compared to normal cells of the same type. A "hypoxic" cell or tissue region, as used herein, refers to a cell or tissue region that is hypoxic or hyperglycolytic, relative to the typical partial pressure of oxygen (pO2) in cells in tissue that is considered normal or healthy. 2 ) lower pO 2 Hypoxic cells or tissue regions can include cells or tissue regions that have reduced vasculature or no access to the vasculature, such as in solid tumors.

[0126] The compounds of the present invention exert their anti-cancer and anti-proliferative effects through a novel mechanism that induces cytostatic activity at concentrations typically 100-1000 times higher than cytostatic concentrations, with less toxicity and cytotoxic or cytocidal effects. The compounds of the present invention induce endoplasmic reticulum stress in the first cytostatic phase, which coincides with ATP and lactate decline in cancer cell cultures. This unique mechanism exhibited by the compounds of the present invention allows for a certain therapeutic window, i.e., an effective dose of the compounds of the present invention that can be used to stop cancerous growth without exerting cytotoxic side effects in the same patient.

[0127] In a preferred embodiment in combination with any of the above or below embodiments, said compound or said pharmaceutical composition comprises: with one or more therapeutic agents for cancer selected from the group consisting of PD-1 agents, PD-L1 agents, CTLA-4 agents, IDO1 inhibitors, and anti-cancer vaccines; or - in conjunction with cytokine therapy or known chemotherapy or drug therapy, or during radiation therapy; is administered.

[0128] Preferred agents, inhibitors, vaccines and treatments according to this embodiment are described below.

[0129] A further aspect of the invention relates to a compound according to any one of the above embodiments (i.e. according to formula (I)) or a pharmaceutical composition comprising a compound according to any one of the above embodiments (i.e. according to formula (I)) for use in a method for preventing and / or treating an autophagy-related disease.

[0130] A further aspect of the present invention relates to a method for preventing and / or treating an autophagy-related disease, comprising administering to a subject in need thereof an effective amount of a compound according to any one of the above embodiments (i.e. according to formula (I)) or a pharmaceutical composition comprising a compound according to any one of the above embodiments (i.e. according to formula (I)).

[0131] The present invention further relates to the use of a compound according to any one of the above embodiments (i.e. according to formula (I)) or a pharmaceutical composition comprising a compound according to any one of the above embodiments (i.e. according to formula (I)) in the manufacture of a medicament for preventing and / or treating an autophagy-related disease.

[0132] In an embodiment in combination with any of the above or below embodiments, said autophagy related disease is preferably selected from the group consisting of neurodegenerative diseases, such as Parkinson's disease, Alzheimer's disease and Huntington's disease; amyotrophic lateral sclerosis; metabolic disorders, such as obesity and diabetes mellitus types I and II; viral infections and diseases leading to the accumulation of certain debris particles, such as fibrotic lung diseases.

[0133] Combinations of Compounds of the Invention with Biguanides As described herein, the compounds according to formula (I) exhibit advantageous utility in the treatment of certain diseases, particularly cancer.In addition, it has been surprisingly found that the compounds according to formula (I) also exhibit unexpectedly advantageous utility in combination with biguanide compounds in the treatment of certain diseases, particularly cancer.

[0134] As explained herein, biguanide compounds such as metformin, phenformin and buformin are known for their activity in the treatment of diabetes and cancer, however, certain of these compounds (particularly buformin and potent phenformin) cause toxic lactic acidosis at higher doses, effectively limiting the dose at which the biguanides can be administered to below that required for therapeutic efficacy, particularly in the treatment of cancer.

[0135] At the same time, as noted above, the compounds of the present invention according to formula (I) themselves exhibit anti-cancer activity that, without being bound by theory, is believed to be related at least in part to the ability of these compounds to reduce lactate levels.

[0136] It has now surprisingly been found that combination therapy comprising administering a compound according to formula (I) of the present invention together with a biguanide compound makes it possible to exploit the therapeutic potential of each of these compounds while simultaneously reducing or eliminating the adverse side effect of lactic acidosis that has hitherto limited the application of biguanides at the higher doses that are typically of concern for the treatment of diseases, in particular for the treatment of cancer. Such combinations of the compounds of the present invention (i.e. according to formula (I)) with biguanide compounds therefore broaden the therapeutic window for the safe administration of the biguanide compound and synergistically exploit the activity of each of these compounds in the prevention and / or treatment of diseases, in particular in the prevention and / or treatment of cancer.

[0137] Thus, a further aspect of the invention relates to a composition comprising a compound according to formula (I) and a biguanide. The biguanide may preferably be metformin, phenformin and / or buformin. The composition may preferably be a pharmaceutical composition comprising, in addition to the inventive compound according to formula (I) and a biguanide, a pharma- ceutically acceptable formulation excipient as specified herein. The composition may be in any form, but preferred forms are those suitable for oral administration in the prophylaxis and / or treatment as specified elsewhere herein.

[0138] A further aspect of the present invention relates to a kit comprising a compound according to formula (I) and a biguanide. The kit may contain the compound according to formula (I) and the biguanide in separate containers. When separate containers, these containers may be of the same or different types that are most suitable for the intended delivery route of the substances. The compound according to formula (I) and the biguanide contained in the kit may be in the same or different delivery forms, i.e., may be present in the kit in the same or different formulations. For example, the compound according to formula (I) contained in the kit may be in a formulation suitable for intravenous delivery as described herein (e.g., a solution in a container fitted with a septum), while the biguanide contained in the same kit may be in a formulation suitable for oral delivery as described herein (e.g., a tablet or capsule contained in a blister pack).

[0139] A further aspect of the invention relates to a composition comprising a compound according to formula (I) and a biguanide or a kit comprising a compound according to formula (I) and a biguanide for use as a medicine. A related aspect of the invention relates to a method of preventing and / or treating a disease comprising administering to a subject a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide combination. A further related aspect of the invention relates to a combination of a compound according to formula (I) and a biguanide for use in therapy. The combination of these substances in the prevention and / or treatment of diseases is associated with the advantages identified herein.

[0140] As described in further detail herein, the combination therapy with the compound according to formula (I) and biguanides is particularly suitable for the treatment of cancer, but such combination therapy may also have similar utility when preventing and / or treating other diseases.For example, this combination therapy may also be applied to the prevention and / or treatment of diseases such as type II diabetes mellitus (T2DM); for example dementia, particularly neurodegenerative diseases such as Alzheimer's dementia, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis (ALS).These substances in the prevention and / or treatment of diseases are associated with the advantages identified herein.Due to the therapeutic potential of this combination therapy in the treatment of cancer, the application of this combination therapy to cancer is discussed in more detail below.

[0141] Thus, a further aspect of the present invention relates to a combination of a compound according to formula (I) and a biguanide, or a kit comprising a compound according to formula (I) and a biguanide, for use in a method for preventing and / or treating cancer. These substances, i.e. the combination of a compound according to formula (I) and a biguanide, are associated with the advantages identified herein in the prevention and / or treatment of cancer. Preferably, the cancer to be prevented and / or treated is an advanced stage cancer, in particular an advanced stage glycolytic cancer. Suitable doses of the compound according to formula (I) of the present invention in this combination therapy are as indicated elsewhere herein. In the case of combination therapy with a compound according to formula (I) of the present invention with metformin as the biguanide, said metformin is advantageously administered in a dose of about 500 mg to about 2000 mg, two, three or four times a day. In such cases, the administration of metformin is preferably via the oral route. In the case of combination therapy with a compound according to formula (I) of the present invention with phenformin as the biguanide, the compound is advantageously administered in a dose of about 25 mg to about 100 mg, two or three times a day. In such a case, administration of phenformin is preferably via the oral route. In the case of combination therapy with a compound according to formula (I) of the present invention with buformin as the biguanide, the compound is advantageously administered in a dose of about 50 mg to about 1000 mg, two or three times a day. In such a case, administration of buformin is preferably via the oral route.

[0142] In a further aspect, the present invention relates to a method for preventing and / or treating cancer, comprising administering to a subject a combination of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide. These substances, i.e. the combination of a compound according to formula (I) and a biguanide, are associated with the advantages identified herein in the prevention and / or treatment of cancer. Preferably, the cancer to be prevented and / or treated is an advanced stage cancer, in particular an advanced stage glycolytic cancer. Suitable doses of the compound according to formula (I) of the present invention in this combination therapy are as indicated elsewhere herein. In the case of combination therapy with a compound according to formula (I) of the present invention with metformin as the biguanide, said metformin is advantageously administered in a dose of about 500 mg to about 2000 mg, two, three or four times a day. In such cases, administration of metformin is preferably via the oral route. In the case of combination therapy with a compound according to formula (I) of the present invention with phenformin as the biguanide, the compound is advantageously administered in a dose of about 25 mg to about 100 mg, two or three times a day. In such a case, administration of phenformin is preferably via the oral route. In the case of combination therapy with a compound according to formula (I) of the present invention with buformin as the biguanide, the compound is advantageously administered in a dose of about 50 mg to about 1000 mg, two or three times a day. In such a case, administration of buformin is preferably via the oral route.

[0143] In a further aspect, the present invention relates to the use of a combination of a compound according to formula (I) or a pharmaceutical composition comprising said compound according to formula (I) and a biguanide in the manufacture of a medicament for preventing and / or treating cancer. These substances, i.e. the combination of a compound according to formula (I) and a biguanide, are associated with the advantages specified herein in the prevention and / or treatment of cancer. Preferably, the cancer to be prevented and / or treated is an advanced stage cancer, in particular an advanced stage glycolytic cancer. Suitable doses of the compound according to formula (I) of the present invention in this combination therapy are as indicated elsewhere herein. In the case of combination therapy with a compound according to formula (I) of the present invention with metformin as the biguanide, said metformin is advantageously administered in a dose of about 500 mg to about 2000 mg, two, three or four times a day. In such cases, the administration of metformin is preferably oral. In the case of combination therapy with a compound according to formula (I) of the present invention and phenformin as the biguanide, the compound is advantageously administered at a dose of about 25 mg to about 100 mg, two or three times a day. In such a case, phenformin administration is preferably oral. In the case of combination therapy with a compound according to formula (I) of the present invention and buformin as the biguanide, the compound is advantageously administered at a dose of about 50 mg to about 1000 mg, two or three times a day. In such a case, buformin administration is preferably oral.

[0144] In a preferred embodiment of the above combination prevention and / or treatment involving a compound according to formula (I) of the invention together with a biguanide, when the disease to be prevented and / or treated is cancer, the cancer may preferably be an LKB-1-deficient cancer. In the present specification, an LKB-1-deficient cancer is understood as a human cancer harboring a mutation, deletion or translocation of the skt11 gene encoding LKB-1 (liver kinase (B1), see, for example, UNIPROT accession number Q15831, and / or NCBI accession number NP_000446), which reduces or eliminates its expression. In one preferred embodiment, the LKB-1-deficient cancer may be one or more sub-forms of skt11 / LKB-1-mutated non-small cell lung cancer (NSCLC, see Facchinetti et al, Lung Cancer. 112:62-68. (2017)). Other cancer types that frequently show mutations, deletions or translocations of LKB-1 function are sarcomas, cholangiocarcinoma, breast adenoid cystic carcinoma, and certain gynecological cancers such as cervical, ovarian or uterine carcinoma (see Li et al., Biomed Pharmacother. 132:110872. (2020)).

[0145] In a further preferred embodiment of the above combination prevention and / or treatment with a compound according to formula (I) of the invention together with a biguanide, when the disease to be prevented and / or treated is cancer, the cancer may preferably be a cancer harboring a mutation, deletion or translocation of arid1, which encodes Arid1a. ARID1a is a gene encoding AT-rich interaction domain-containing protein 1A in humans (see, for example, UNIPROT accession number O14497 and / or NCBI accession numbers NP_006006 and NP_624361). The elevated proteotoxic stress through genetic damage of Arid1a together with pharmacological treatments resulting in further increased proteotoxic stress responses generally gives a kind of "synthetic lethality" situation, which is an excellent requirement for preferred pharmacological treatments (Huang et al., Nat Rev Drug Discov 19(1):23-38(2020)).

[0146] In a particularly preferred embodiment of the above combination prevention and / or treatment involving a compound according to formula (I) of the present invention together with a biguanide, when the disease to be prevented and / or treated is cancer, the cancer may preferably be simultaneously LKB-1-deficient and ARID1a-mutated cancer. Without being bound by theory, such double deletion / mutation cancers are particularly suitable for treatment by combination therapy of a compound according to formula (I) of the present invention and a biguanide, because the additional mutation in ARID1a in cancers that already lack LKB-1 makes LKB-1-deficient cancers more susceptible to proteotoxic stress. In a particularly preferred embodiment, one such cancer harboring mutations in both LKB-1 and ARID1a is non-small cell lung cancer.

[0147] (definition) All technical and scientific terms used below have the commonly understood meaning of the term unless otherwise defined.

[0148] It should be understood that the section headings used throughout this application are merely for organizational purposes and are not intended to confer an exclusive character on the disclosure content related to the aspect of the invention referred to in the section heading. As such, the section headings do not preclude the combination of the disclosure content within any one section heading for a corresponding or similar context being provided under another section heading. The disclosure content within such sections is expressly within the scope of the disclosure content of this application.

[0149] As used herein, each of the terms "comprising," "having," and "containing," including its grammatical variations, is intended in an open sense to mean "including," but not necessarily "composed of," and does not exclude elements in addition to those specifically recited as being present. As used herein, the terms "comprising," "having," and "containing," and their grammatical variations, indicate that components other than those specifically recited may, but need not, be present. Thus, these terms include, for example, the commonly accepted meaning of "consisting of," as a limiting example, an embodiment in which no elements other than those recited are present.

[0150] As used herein, the phrase "consisting of" and its grammatically related variations mean that no other elements are present in the recited set. In support of the above definition of "comprising" (and grammatically and semantically related terms), the term "consisting of" therefore represents a limiting scenario within the meaning of "comprising."

[0151] Unless otherwise defined, any feature within any aspect or embodiment of the present invention may be combined with any feature within any other aspect or embodiment of the present invention, and those skilled in the art will understand that such combinations are encompassed by the inherent disclosure of this application. This applies in particular to all embodiments described within the section relating to the compounds themselves, as well as to other aspects, such as the methods and uses of these compositions. This also applies in particular, but not exclusively, to the end points of ranges disclosed herein. For example, when a given substance is disclosed as being present in a composition within a concentration range of about XY% or about AB%, the application should be understood to expressly disclose not only the ranges of about XY% and about AB%, but also the ranges of about XB%, about AY%, and, where numerically possible, about YA% and about BX%. Each of these ranges and combinations of ranges are contemplated and should be understood to be directly and explicitly disclosed in this application.

[0152] Unless otherwise stated, any expression of a range in this application using two bracket values ​​X and Y, or a hyphen ("-") separating two bracket ratios, should be understood to mean and disclose the particular range in which both endpoint values ​​X and Y are included. The same applies to ranges expressed as "from about X to about Y". Thus, expressions of ranges such as "XY", "X-Y", "X to Y", "XY" and "X to (-)Y" should be understood equivalently to mean and disclose the range including the end value X, all values ​​between X and Y, and the end value Y. In the event that the range in question specifies the number of atoms in an entity where only integer values ​​can have technical meaning, e.g., the term "C 1 -C 6 Alkyl" or equivalently "C 1-6 In the case of a chemical substituent such as "alkyl," the ranges specified above are understood to mean and disclose each integer value within that range. For example, "C 1 -C 6 Alkyl" (or equivalently "C 1-6 In the example of "alkyl", the term is divided into separate integer options "C1 Alkyl, C 2 Alkyl, C 3 Alkyl, C 4 Alkyl, C 5 Alkyl" and "C 6 It should be understood that the term "alkyl" refers to and directly and explicitly discloses each of the terms "alkyl" and "alkyl-alkyl."

[0153] Any range expression in this application using the word "between" preceding two bracket values ​​X and Y, or two bracket ratios, should be understood to mean and disclose the specified range in which both endpoint values ​​X and Y are excluded, but all values ​​between the specified endpoint values ​​X and Y are included. As used herein, the term "about" when referring to a particular value, for example, to an endpoint(s) of a range, encompasses and discloses a certain variation about the specifically recited value in addition to the specifically recited value itself. Such variation may result, for example, from normal measurement variations in weighing or dispensing various substances by methods known to those skilled in the art. The term "about" is understood to encompass and disclose a range of variation above and below the particular value indicated, and the percentage values ​​above are relative to the particular recited value itself, as follows: The term "about" may represent a variation of ±5.0%. The term "about" may represent a variation of ±4.9%. The term "about" may represent a variation of ±4.8%. The term "about" may represent a variation of ±4.7%. The term "about" may represent a variation of ±4.6%. The term "about" may represent a variation of ±4.5%. The term "about" may represent a variation of ±4.4%. The term "about" may represent a variation of ±4.3%. The term "about" may represent a variation of ±4.2%. The term "about" may represent a variation of ±4.1%. The term "about" may represent a variation of ±4.0%. The term "about" may represent a variation of ±3.9%. The term "about" may represent a variation of ±3.8%. The term "about" may represent a variation of ±3.7%. The term "about" may represent a variation of ±3.6%. The term "about" may represent a variation of ±3.5%. The term "about" may represent a variation of ±3.4%. The term "about" may represent a variation of ±3.3%. The term "about" may represent a variation of ±3.2%. The term "about" may represent a variation of ±3.1%. The term "about" may represent a variation of ±3.0%. The term "about" may represent a variation of ±2.9%. The term "about" may represent a variation of ±2.8%. The term "about" may represent a variation of ±2.7%. The term "about" may represent a variation of ±2.6%. The term "about" may represent a variation of ±2.5%. The term "about" may represent a variation of ±2.4%. The term "about" may represent a variation of ±2.3%. The term "about" may represent a variation of ±2.2%.The term "about" may represent a variation of ±2.1%. The term "about" may represent a variation of ±2.0%. The term "about" may represent a variation of ±1.9%. The term "about" may represent a variation of ±1.8%. The term "about" may represent a variation of ±1.7%. The term "about" may represent a variation of ±1.6%. The term "about" may represent a variation of ±1.5%. The term "about" may represent a variation of ±1.4%. The term "about" may represent a variation of ±1.3%. The term "about" may represent a variation of ±1.2%. The term "about" may represent a variation of ±1.1%. The term "about" may represent a variation of ±1.0%. The term "about" may represent a variation of ±0.9%. The term "about" may represent a variation of ±0.8%. The term "about" may represent a variation of ±0.7%. The term "about" may represent a variation of ±0.6%. The term "about" may represent a variation of ±0.5%. The term "about" may represent a variation of ±0.4%. The term "about" may represent a variation of ±0.3%. The term "about" may represent a variation of ±0.2%. The term "about" may represent a variation of 0.1%. The term "about" may represent the exact specific value itself with respect to a particular recited value, notwithstanding any express reference that this exact value is included; even in the absence of an express indication that the term "about" includes the exact specific recited value, this exact specific value is still included within the range of variation created by the term "about" and is hereby disclosed. Unless otherwise stated, when the term "about" is recited before the first endpoint of a numerical range, the term refers to both the first endpoint of the range and the second endpoint of the range. For example, a recited range "from about X to Y" should be read as "from (about) X to about Y."

[0154] As used herein, "C 1-6 By "alkyl" is meant a saturated alkyl chain having 1, 2, 3, 4, 5 or 6 carbon atoms, which may be linear or branched. In the context of the present invention, the term "C 1-6 C in the "Alkyl" range 1-3 Alkyl, C 2-5 Alkyl or C 3-6 Any subgroups such as alkyl groups are also included. 1-6Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, and hexyl.

[0155] "C 1-6 "Alkylene" means that each group is divalent and connects the remainder of the bond to the rest of the molecule. 1 Alkylene means a methylene linker, C 2 Alkylene means an ethylene linker or a methyl-substituted methylene linker, etc. In the context of the present invention, 1-6 Alkylene preferably denotes a methylene or ethylene group.

[0156] The term “OC 1-6 "Alkyl" means that the alkyl chain is attached to the rest of the molecule through an oxygen atom.

[0157] The term "Halo-C 1-6 "Alkyl" means that one or more hydrogen atoms in the alkyl chain are replaced by halogen. A preferred example thereof is CF 3 O-Halo-C 1-6 Alkyl means an alkoxy group having one or more hydrogen atoms replaced by halogen.

[0158] C 3-6 Cycloalkyl group means a saturated or partially unsaturated mono- or bicyclic ring system containing 3, 4, 5 or 6 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cyclohexenyl.

[0159] A 4-8 membered monocyclic or bicyclic heterocycloalkyl group has up to four carbon atoms replaced by up to four heteroatoms, and the heteroatoms are N, O, S, SO and SO. 2means a saturated or partially unsaturated 4-8 membered monocyclic or bicyclic carbocyclic ring independently selected from: Examples include epoxydyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, 1,4-dioxanyl, morpholinyl, 4-quinuclidinyl, 1,4-dihydropyridinyl, and 6-azabicyclo[3.2.1]octanyl. The heterocycloalkyl group may be attached to the remainder of the molecule through a carbon, nitrogen (e.g., in morpholine or piperidine), or sulfur atom.

[0160] A cycloalkyl or heterocycloalkyl substituent may be spirocyclically fused to the rest of the molecule. In the context of this invention, "spirocyclically fused" means that a cycloalkyl or heterocycloalkyl group is connected to the rest of the molecule through a single common atom, i.e., the ring shares a common atom with another moiety. Examples of spiro connections are shown below: [ka]

[0161] A 5- or 6-membered heteroaromatic ring system containing up to four heteroatoms (also referred to herein as heteroaryl) means a monocyclic heteroaromatic ring such as pyrrolyl, imidazolyl, furanyl, thiophenyl, pyridinyl, pyrimidinyl, pyrazinyl, pyrazolyl, oxazolyl, isoxazolyl, triazolyl, oxadiazolyl and thiadiazolyl.

[0162] The nitrogen atom of a heteroaryl system may be optionally oxidized to the corresponding N-oxide. Furthermore, the nitrogen atom of said heteroaryl system may be optionally quaternized. Unless otherwise stated, said heteroaryl system may be attached via a carbon or nitrogen atom.

[0163] Additionally, unless expressly defined, the term "heteroaryl" contains 1 to 4 heteroatoms independently selected from the group consisting of N, O and S.

[0164] A six-membered aromatic ring system (also referred to within this application as aryl) means an aromatic carbocyclic ring such as phenyl.

[0165] The above structural parts [ka] It should be understood to represent the 8 or 9 membered bicyclic heteroaryl rings exemplified, but not limited to, below. [ka]

[0166] Structural moiety in formula (II) [ka] It should be understood to represent a 9-membered bicyclic heteroaryl ring, exemplified but not limited by the following: [ka]

[0167] The term "halogen" includes the specific halogen atoms fluorine, bromine, chlorine and iodine.

[0168] It will be understood by those skilled in the art that when a list of alternative substituents includes members that cannot be used to replace a particular group because of their valency requirements or for other reasons, said list is intended to include only those members of said list that, as read by the knowledge of one of skill in the art, are suitable to replace the particular group without contradicting the valency rules.

[0169] Any formula or structure given herein is intended to represent the unlabeled form of the above compound as well as the isotopically labeled form. Isotopically labeled compounds have a structure represented by the formula given herein, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds of the present disclosure include, but are not limited to: 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 15 N, 18 F, 35 S, 36 Cl and 125 These include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, such as I. The present invention therefore provides for the synthesis of such isotopically labeled compounds, e.g. 3 H, 13 C and 14 These include those containing a radioisotope such as C. Such isotopically labeled compounds may be useful in detection or imaging techniques such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including metabolism studies, reaction kinetic studies, drug or substrate tissue distribution assays in patients or radiation treatment. Isotopically labeled compounds of the present invention can generally be prepared by carrying out the procedures disclosed in the schemes or examples and preparations described below by substituting readily available isotopically labeled reagents for non-isotopically labeled reagents.

[0170] The present invention also includes "deuterated analogs" of compounds of formula (I) in which one to n hydrogens attached to a carbon atom are replaced by deuterium, where n is the number of hydrogens in the molecule. Such compounds may exhibit increased resistance to metabolism and therefore may be useful for increasing the half-life of any compound of formula (I) when administered to a mammal, e.g., a human. See, e.g., Foster in Trends Pharmacol.Sci.1984:5;524. Such compounds are synthesized by means well known in the art, e.g., by employing starting materials in which one or more hydrogens have been replaced by deuterium, or are enriched in deuterium at the position(s) of interest.

[0171] Deuterium-labeled or substituted therapeutic compounds of the invention may have improved DMPK (drug metabolism and pharmacokinetic) properties, which relate to distribution, metabolism and excretion (ADME). Substitution with heavier isotopes, such as deuterium, may confer certain therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life, reduced dosing requirements and / or improved therapeutic index. 18 F-labeled compounds may be useful for PET or SPECT studies.

[0172] The concentration of such heavier isotopes, specifically deuterium, can be defined by the isotopic enrichment factor. In the compounds of the present invention, any atom not specifically designated as a particular isotope is intended to represent any stable isotope of that atom. Unless otherwise stated, when a position is specifically designated by "H" or "hydrogen", it is understood that the position has hydrogen at its natural abundance in its isotopic composition.

[0173] Metabolites of the compounds of the invention are also within the scope of the invention.

[0174] In cases where tautomerism, such as, for example, keto-enol tautomerism, of the compounds of the invention may occur, the individual forms, such as, for example, keto and enol forms, are each within the scope of the invention as well as mixtures in any ratio. The same applies to stereoisomers, such as, for example, enantiomers, cis / trans isomers, conformers, etc.

[0175] If desired, isomers can be separated by methods well known in the art, for example by liquid chromatography. The same applies to enantiomers, for example by using chiral stationary phases. In addition, enantiomers can be isolated by converting them to diastereomers, i.e. by coupling with an enantiomerically pure auxiliary compound, followed by separation of the resulting diastereomers and cleavage of the auxiliary residue. Alternatively, any enantiomer of the compound of the present invention can be obtained from stereoselective synthesis using optically pure starting materials. Another method of obtaining pure enantiomers from racemic mixtures can use enantioselective crystallization with chiral counterions, by methods well known in the art.

[0176] As used herein, “metformin” refers to a compound having the following chemical structure: [ka]

[0177] As used herein, "phenformin" refers to a compound having the following chemical structure: [ka]

[0178] As used herein, “buformin” refers to a compound having the following chemical structure: [ka]

[0179] Administration of one or more compounds of the invention "together with" or "in combination with" another substance(s) or type of treatment, as used herein, does not necessarily represent simultaneous administration. Rather, a "combination" of two or more compounds, such as administration of one compound "together with" or "in combination with" another or other or type of treatment, as used herein, encompasses and discloses multiple possibilities regarding the route and timing of administration of each substance and / or treatment. Included and disclosed in this regard are, for example, administration of two substances simultaneously by the same route, simultaneous administration by different routes, administration alternating over time by the same route, or administration alternating over time by different routes. Any of the administration routes described herein may be combined in any way. Moreover, the order of administration is not important. For example, applying the general example of two substances A and B as the administration of substances A and B "along with" or "in combination with" each other, alternating over time, or as a "combination", may include the administration of substance A first and substance B later, or the administration of substance B first and substance A later. For example, the administration of substances A and B alternating by the same route may take the form of first oral administration of substance A followed by oral administration of substance B; the administration of substances A and B thus falls within the meaning of, and is disclosed by, the administration of the two substances "along with" or "in combination with" each other. Similarly, the administration of substances A and B alternating by different routes may take the form of first oral administration of substance A followed by intravenous administration of substance B; the administration of substances A and B thus also falls within the meaning of, and is disclosed by, the administration of the two substances "along with" or "in combination with" each other. The administration of substances A and B in one and the same formulation is also envisioned by the present invention. In some embodiments, particularly those relating to combinations of a compound of the invention according to Formula (I) and a biguanide, the route of administration is preferably oral and the timing of administration of the combined agents is preferably simultaneous.In other embodiments, particularly relating to the combination of a compound of the invention according to formula (I) and a biguanide, the route of administration is preferably intravenous for the compound according to formula (I) and oral for the biguanide, in which case the administration of each of the combined substances may be timed to be simultaneous or staggered over time.

[0180] As intended herein, there is no particular limit as to the time period between the administration of two agents "together with" or "in combination with" each other, so long as the two agents in question are administered as part of the same overall prophylactic and / or therapeutic regimen related to the disease or condition in question.

[0181] Similarly, administration of an agent "during" a specified type of treatment, as used herein, should not be understood as requiring simultaneous over time. Rather, administration of an agent(s) "during" a treatment encompasses and includes any administration timing and route as part of a broader prophylactic and / or therapeutic regimen. For example, administration of one or more compounds according to the present invention "during" a subject's radiation therapy does not require, but includes the possibility, that such compound(s) be administered to the subject at the same time that radiation is being administered to the subject. For example, administration "during" a radiation therapy can include administration of one or more compounds of the present invention before or after such radiation application, or before and after such radiation application, or before, simultaneously with and after such radiation application.

[0182] As used herein, the term "advanced stage cancer" should be understood to mean cancer classified as stage III or above according to the TMN staging system (see American Society for Clinical Oncology (ASCO) definitions, accessible under https: / / www.cancer.net / navigating-cancer-care / diagnosing-cancer / stages-cancer).

[0183] As used herein, the term "glycolytic cancer" refers to cancers characterized by above average glucose consumption, i.e., 19 as evidenced by F-deoxyglucose inoculation, and 1 It should be understood as meaning a cancer that presents above average lactate production, demonstrated by H-NMR or by determining blood lactate levels by enzymatic analysis.

[0184] As used herein, the term "endoplasmic reticulum stress" should be understood to mean a response originating from the endoplasmic reticulum characterized by certain markers, such as upregulation of Chac and Chop at the transcriptional level. The above term may be understood to have the same meaning as the term unfolded protein response (see Thangaraj et al. International Review of Cell and Molecular Biology. Vol 350, 285-325 (2020)).

[0185] As used herein, the terms "reduction of hypoxic areas" and "significant reduction of hypoxic areas" should be understood to mean the disappearance of pimonidazole positive staining areas in tissue sections as evidenced by immunofluorescence staining and subsequent morphometric analysis. A significant reduction exists when quantitative image analysis of at least 10 individual sections and stainings shows a statistically significant reduction in pimonidazole positive staining compared to the respective control groups analyzed by appropriate statistical methods (see Zaidi et al., Front Bioeng Biotechnol. 5;7:397. (2019)).

[0186] The compounds of the present invention may be in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids, including inorganic bases or acids and organic bases or acids. When the compounds of the present invention contain one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically or toxicologically acceptable salts, in particular their pharmaceutically usable salts. Thus, the compounds of the present invention that contain acidic groups may exist in these groups and be used according to the present invention, for example, as alkali metal salts, alkaline earth metal salts or ammonium salts. More specific examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts, or salts with ammonia, or organic amines, such as, for example, ethylamine, ethanolamine, triethanolamine, or amino acids. The compounds of the present invention contain one or more basic groups, i.e., groups that can be protonated, and may exist and be used according to the present invention in the form of their addition salts with inorganic or organic acids. Examples of suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to those skilled in the art. When the compounds of the present invention contain both acidic and basic groups in the molecule, the present invention also includes inner salts or betaines (zwitterions) in addition to the salt forms mentioned. The respective salts mentioned above can be obtained by conventional methods known to those skilled in the art, for example by contacting them with organic or inorganic acids or bases in a solvent or dispersant, or by anion or cation exchange with other salts. The present invention also includes all salts of the compounds of the invention which, due to their low physiological compatibility, are not directly suitable for pharmaceuticals but can be used, for example, as intermediates in chemical reactions or in the preparation of pharma-ceutically acceptable salts.

[0187] Further compounds of the present invention may exist in the form of pharma- ceutically acceptable solvates, such as those that include water of solvation, or with alcohols, in particular ethanol.

[0188] The following now describe exemplary, non-limiting ways in which a compound according to formula (I) may be formulated for administration to a subject. It should be understood that throughout the following, the possible formulations are intended to apply to formulations of the compounds of the invention, i.e., compounds according to formula (I), either alone or in combination with another substance, such as, for example, a biguanide, as described anywhere herein, even if the presence of such other substance is not mentioned.

[0189] Furthermore, when "compounds" of the invention are referred to hereinafter, this should be understood as including the limiting case where a compound of the invention, i.e., a compound according to formula (I) herein, is specified. Similarly, reference to an "active compound" or "active compounds" also refers to a compound of the invention, i.e., a compound according to formula (I) herein. Similarly, reference to an "invention compound" or "invention compounds" also refers to a compound of the invention, i.e., a compound according to formula (I) herein.

[0190] Furthermore, the present invention provides a pharmaceutical composition comprising at least one compound of the present invention, or a pharma- ceutically acceptable salt or solvate thereof, as an active ingredient, together with a pharma- ceutically acceptable carrier, which also applies in the event that at least one compound of the present invention according to formula (I) is administered, preferably in combination with another compound, such as a biguanide.

[0191] "Pharmaceutical composition" refers to one or more compounds of the present invention and one or more inactive ingredients that make up the carrier, as well as any product that results directly or indirectly from the combination, complex or aggregation of any two or more of the ingredients, or from the dissociation of one or more of the ingredients, or from other types of reactions and interactions of one or more of the ingredients. Thus, the compositions and pharmaceutical compositions of the present invention encompass any composition made by mixing at least one compound of the present invention and a pharma- ceutical acceptable carrier.

[0192] In practice, the compounds of the present invention can be combined as active ingredients in intimate admixture with pharmaceutical carriers by conventional pharmaceutical compounding techniques. The carriers can take a wide variety of forms depending on the preparation form desired for administration, e.g., oral or parenteral (including intravenous). In preparing compositions for oral dosage forms, any of the usual pharmaceutical media can be employed, e.g., for oral liquid preparations such as suspensions, elixirs and solutions, e.g., water, glycols, oils, alcohols, flavorings, preservatives, colorings, etc.; or for oral solid preparations such as powders, hard and soft capsules and tablets, e.g., starches, sugars, microcrystalline cellulose, diluents, gelatinizing agents, lubricants, binders, disintegrants, etc., and the solid oral preparations are preferred over the liquid preparations.

[0193] For ease of administration, tablets and capsules represent the most advantageous oral dosage unit forms, in which case solid pharmaceutical carriers are naturally employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques. Such compositions and preparations should contain at least about 0.1 percent of the active compound. The percentage of the active compound in these compositions may, of course, vary and may conveniently be between about 5 percent and about 98 percent of the weight of the unit, more preferably between about 10 and about 90 percent of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that an effective dosage is obtained. The active compound may also be administered intranasally, for example, as drops or spray. The tablets, pills, capsules, etc. may also contain binders such as xanthan gum, acacia, cornstarch, or gelatin; formulation excipients such as dicalcium phosphate; disintegrating agents such as cornstarch, potato starch, alginic acid; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose, or saccharin. When the dosage unit form is a capsule, it may contain, in addition to material of the above type, a liquid carrier such as a fatty oil.

[0194] Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye, and a flavoring such as cherry or orange flavor.

[0195] The compounds used in the present invention may be administered parenterally.These active compounds can be prepared in water, suitably mixed with a surfactant such as hydroxy-propylcellulose.Dispersions can also be prepared in oil, glycerol, liquid polyethylene glycols and mixtures thereof.Under normal storage and use conditions, these preparations contain a preservative to prevent the growth of microorganisms.

[0196] Suitable pharmaceutical forms for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and fluid to the extent that easy syringability remains. The form must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be, for example, a solvent or dispersion medium containing water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0197] More specific details regarding these and further modes of formulation are set forth below.

[0198] Formulation of the invention compound suitable for oral administration: Compounds of the invention suitable for oral administration may be prepared or packaged in the form of discrete solid dosage units, including sachets (e.g., powders in sachets), tablets, hard or soft capsules, cachets, troches, or lozenges, each containing a predetermined amount of the compound(s) of the invention, or may be solid. Other formulations suitable for oral administration include powder or granular formulations, aqueous or oily suspensions, aqueous or oily solutions, or emulsions. As used herein, "oily" liquids include carbon-containing liquid molecules that exhibit less polarity than water.

[0199] Tablets containing the compounds of the present invention may be made, for example, by compressing or molding the invention compound, optionally with one or more additional ingredients. Compressed tablets may be prepared by compressing the invention compound in a free-flowing form, such as a powder or granular preparation, optionally with one or more of a binder, lubricant, excipient, surfactant, and dispersant in a suitable device. Molded tablets may be made, for example, by molding the invention compound, a pharma- ceutically acceptable carrier, and at least sufficient liquid to moisten the mixture in a suitable device.

[0200] Pharmaceutically acceptable excipients used in the manufacture of tablets include, but are not limited to, inert diluents, gelatinizing and disintegrating agents, binders, and lubricants. Known dispersing agents include, but are not limited to, potato starch and sodium starch glycolate. Known surface active agents include, but are not limited to, sodium lauryl sulfate. Known diluents include, but are not limited to, calcium carbonate, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, calcium hydrogen phosphate, and sodium phosphate. Known gelatinizing and disintegrating agents include, but are not limited to, corn starch and alginic acid. Known binders include, but are not limited to, gelatin, acacia, pregelatinized corn starch, polyvinylpyrrolidone, and hydroxypropyl methylcellulose. Known lubricants include, but are not limited to, magnesium stearate, stearic acid, silica, and talc.

[0201] The tablets may be uncoated or may be coated using known methods to achieve delayed disintegration in the subject's gastrointestinal tract, thereby providing sustained release and absorption of the inventive compound. For example, materials such as glyceryl monostearate or glyceryl distearate may be used to coat the tablets. The tablets may further comprise a sweetener, a flavoring agent, a coloring agent, a preservative, or some combination thereof, to provide a medicamentically attractive and palatable preparation.

[0202] Hard capsules containing the inventive compound may be made using physiologically degradable compositions such as gelatin or cellulose derivatives. Such hard capsules contain the inventive compound and may further contain additional ingredients including, for example, inert solid diluents such as calcium carbonate, calcium phosphate, or kaolin.

[0203] Soft gelatin capsules containing the inventive compound may be made using a physiologically degradable composition such as gelatin combined with a plasticizer (i.e., glycerol) as the basic component of the soft gelatin shell. Soft gelatin capsules may contain liquid or semi-liquid solutions, suspensions, or microemulsion preconcentrates. The filling of the soft capsules contains the inventive compound and may be mixed with water or an oil medium such as peanut oil, liquid paraffin, olive oil, soybean oil, sunflower oil, lecithin such as soybean lecithin or sunflower lecithin, medium chain triglycerides, polyglycerol oleate, beeswax, mono- and diglycerides of fatty acids, or any combination of the above.

[0204] Liquid formulations of the compounds of the invention particularly suitable for oral administration may be prepared, packaged, and sold either in liquid form or in the form of a dry product intended for reconstitution with water or another suitable vehicle prior to ingestion.

[0205] Liquid suspensions may be prepared using conventional methods to achieve suspension of active ingredients in aqueous or oily vehicles. Aqueous vehicles include, for example, water and isotonic saline. Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis oil, olive oil, sesame oil, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Liquid suspensions may further include one or more additional ingredients, including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavoring agents, coloring agents, and sweetening agents. Oily suspensions may further include a thickening agent. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible oils, sodium alginate, polyvinylpyrrolidone, xanthan gum, acacia gum, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, and hydroxypropylmethylcellulose. Known dispersing or wetting agents include, but are not limited to, naturally occurring phospholipids such as lecithin, condensation products of alkylene oxides with fatty acids, long chain aliphatic alcohols, partial esters derived from fatty acids and hexitols, or partial esters from fatty acids and hexitol anhydrides (e.g., polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively). Known emulsifying agents include, but are not limited to, lecithin and acacia. Known preservatives include, but are not limited to, methyl, ethyl, or n-propyl para-hydroxybenzoate, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin. Known thickening agents for oily suspensions include, for example, beeswax, hard paraffin, and cetyl alcohol.

[0206] Powdered or granular formulations of the inventive compounds, e.g., pharmaceutical compositions of the present invention, may be prepared using known methods. Such formulations may be administered directly to a subject, for example, by forming sachets or tablets, filling capsules, or used to prepare aqueous or oily suspensions or solutions by adding aqueous or oily vehicles thereto. Each of these formulations may further comprise one or more of dispersing or wetting agents, suspending agents, and preservatives. Additional formulation excipients, such as fillers and sweetening, flavoring, or coloring agents, may also be included in these formulations.

[0207] The inventive compounds may also be prepared, packaged or sold in the form of oil-in-water emulsions or water-in-oil emulsions. The oil phase may be a vegetable oil such as olive oil or arachis oil, a mineral oil such as liquid paraffin, or a combination thereof. Such compositions may further comprise one or more emulsifiers, such as naturally occurring gums such as gum acacia or xanthan gum, naturally occurring phospholipids such as soybean or lecithin phospholipids, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides, such as sorbitan monooleate and polyoxyethylene sorbitan monooleate, condensation products of such partial esters with ethylene oxide. These emulsions may contain additional ingredients, including, for example, sweeteners or flavoring agents.

[0208] Methods for impregnating or coating materials with chemical compositions are known in the art and include, but are not limited to, depositing or bonding the chemical composition onto a surface, incorporating the chemical composition into the structure of the material during its synthesis (e.g., by means of a physiologically degradable material), and absorbing an aqueous or oily solution or suspension into an absorbent material with or without subsequent drying.

[0209] Formulations of invention compounds suitable for parenteral administration: For parenteral administration, the compounds of the invention may be formulated for injection or infusion, e.g., intravenous, intramuscular or subcutaneous injection or infusion, or for administration in a bolus dose and / or continuous infusion. Suspensions, solutions or emulsions in oily or aqueous vehicles may be used, which may optionally contain other agents such as suspending, stabilizing and / or dispersing agents such as those mentioned above.

[0210] The invention compounds may be made particularly suitable for parenteral administration by formulation with a pharma- ceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus or continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampoules, crushable, or otherwise, or in multi-dose containers containing a preservative. Compositions particularly suitable for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such compositions may further comprise one or more additional ingredients, including, but not limited to, suspending agents, stabilizing agents, or dispersing agents. In one embodiment of the invention composition that is particularly suitable for parenteral administration, the active ingredient is provided in a dry (e.g., powder or granules) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0211] The inventive compound may be prepared in the form of a sterile injectable aqueous or oily suspension or solution. The suspension or solution may be formulated by known techniques and may contain, in addition to the active ingredient described above, additional ingredients such as dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable compositions may be prepared using a non-toxic, parenterally acceptable diluent or solvent, such as water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic saline, and fixed oils such as synthetic mono- or diglycerides. Other conventional parenterally administrable formulations include those containing the active ingredient in microcrystalline form, in liposomal preparations, or as components of biodegradable polymeric systems. Compositions for sustained release or implantation may include pharma-ceutically acceptable polymeric or hydrophobic materials, such as emulsions, ion exchange resins, sparingly soluble polymers, or sparingly soluble salts.

[0212] Formulations of invention compounds suitable for transmucosal administration: The invention compounds may be formulated so that they are suitable for transmucosal administration. The formulation may include any material or dosage unit suitable for application to mucosal tissue. For example, the selected active agent may be administered to the buccal mucosa in an adhesive tablet or patch, sublingually by placing a solid dosage form under the tongue, lingually by placing a solid dosage form under the tongue, intranasally as drops or nasal spray, non-aerosolized liquid formulations, or dry powder, placed in or near the rectum ("rectal" formulation), or administered to the urethra as a suppository, ointment, or the like.

[0213] Formulation of the invention compound suitable for transurethral administration: The invention compounds may be formulated to be suitable for transurethral administration. In this case, the invention compositions may include a urethral dosage form comprising an active agent and one or more selected carriers or excipients, such as water, silicones, waxes, petrolatum, polyethylene glycols ("PEG"), propylene glycols ("PG"), liposomes, sugars such as mannitol and lactose, and / or various other materials. A transurethral permeation enhancer may be included in the dosage form. Examples of suitable permeation enhancers include dimethylsulfoxide ("DMSO"), dimethylformamide ("DMF"), N,N-dimethylacetamide ("DMA"), decylmethyl ("C10MSO"), polyethylene glycol monolaurate ("PEGML"), glycerol monolaurate, lecithin, 1-substituted azacycloheptan-2-ones, particularly 1-n-dodecyl-azacycloheptan-2-one, surfactants as discussed above, e.g., TWEEN-80™, and lower alcohols such as ethanol.

[0214] Formulation of the invention compound suitable for rectal administration: The invention compounds may also be formulated to be suitable for rectal administration. Rectal dosage forms may include rectal suppositories, creams, ointments, and liquid formulations (enemas). Suppositories, creams, ointments, or liquid formulations for rectal delivery include the invention compounds and one or more conventional non-toxic carriers suitable for rectal drug administration. Rectal dosage forms of the invention compositions may be manufactured using conventional processes. Rectal dosage units may be fabricated to disintegrate quickly or over a period of several hours. The period for complete disintegration may be within the range of about 10 minutes to about 6 hours, for example, less than about 3 hours.

[0215] Formulations of the compounds of the invention suitable for intravaginal or transvaginal administration: The inventive compounds may be formulated to be suitable for intravaginal or transvaginal administration. Suitable dosage forms for this purpose may include intravaginal suppositories, creams, ointments, liquid formulations, pessaries, tampons, gels, pastes, foams or sprays. The suppositories, creams, ointments, liquid formulations, pessaries, tampons, gels, pastes, foams or sprays for intravaginal or transvaginal delivery contain a therapeutically effective amount of a selected active agent and one or more conventional non-toxic carriers suitable for intravaginal or transvaginal drug administration. The intravaginal or transvaginal forms of the invention may be manufactured using conventional processes, for example, as disclosed in Remington: The Science and Practice of Pharmacy, supra. The intravaginal or transvaginal dosage units may be fabricated to disintegrate quickly or over a period of several hours. The period for complete disintegration may be within the range of about 10 minutes to about 6 hours, for example, less than about 3 hours.

[0216] Formulation of the invention compound suitable for topical application: The inventive compounds may be formulated to be suitable for topical administration. Formulations suitable for this purpose may include any form suitable for application to body surfaces, including, for example, ointments, creams, gels, lotions, solutions, pastes, etc., and / or may be prepared to contain liposomes, micelles, and / or microspheres. In certain embodiments, the topical formulations herein are ointments, creams, and gels.

[0217] Formulation of the invention compound suitable for transdermal administration: The inventive compound may be formulated to be particularly suitable for transdermal administration. As known to those skilled in the art, transdermal administration involves the delivery of a pharmaceutical compound to the patient's systemic circulation via the transdermal route of the compound. This may be done, for example, by a transdermal patch or an iontophoretic device. Other ingredients besides the inventive compound may also be incorporated into the transdermal patch. For example, the composition and / or transdermal patch may be formulated with one or more preservatives or bacteriostatic agents, including, but not limited to, methyl hydroxybenzoate, propyl hydroxybenzoate, chlorocresol, benzalkonium chloride, and the like. The dosage form of the inventive compound may include creams, sprays, lotions, gels, ointments, eye drops, nose drops, ear drops, and the like. In such dosage forms, the components of the inventive composition may be mixed to form a white, smooth, opaque cream or lotion, for example with 1% or 2% (wt / wt) benzyl alcohol, emulsifying wax, glycerin, isopropyl palmitate, lactic acid, purified water and sorbitol solution as preservatives. The composition may also contain polyethylene glycol 400. These may be mixed to form an ointment, for example with about 2% (wt / wt) benzyl alcohol, white petrolatum, emulsifying wax and tenox II (butylated hydroxyanisole, propyl gallate, citric acid, propylene glycol) as preservatives. Dressings, for example woven pads or rolls of gauze, may be impregnated with the composition in a solution, lotion, cream, ointment, etc., and such forms may be used for topical application. The composition may be applied topically using a transdermal system, such as one of an acrylic polymer adhesive with a resinous crosslinker impregnated with the composition and laminated to an impermeable backing.

[0218] Examples of suitable skin contact adhesive materials include, but are not limited to, polyethylene, polysiloxane, polyisobutylene, polyacrylate, polyurethane, and the like. Alternatively, the drug-containing reservoir and the skin contact adhesive are separate and distinct layers, with the adhesive under the reservoir, which in this case may be either a polymer matrix as described above, or a liquid or hydrogel reservoir, or take some other form. Any suitable route of administration may be employed to provide an effective dose of the compound of the present invention to a mammal, particularly a human. For example, oral, rectal, topical, parenteral (including intravenous, intramuscular, and subcutaneous), ocular (ocular), pulmonary (nasal or oral inhalation), nasal, and the like may be employed. As described above, dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, creams, ointments, aerosols, and the like. Preferably, the compound of the present invention is administered orally or intravenously.

[0219] The effective dosage of the active ingredient employed may vary depending on the particular compound employed, the mode of administration, the condition being treated, and the severity of the condition being treated. Such dosages can be readily ascertained by one skilled in the art.

[0220] When the compounds of formula (I) are used to treat or prevent lactate / ATP-mediated diseases, generally satisfactory results are obtained when the compounds are administered in a daily dosage of about 0.1 mg to about 100 mg per kilogram of mammalian body weight, preferably given as a single dose or in divided doses 2 to 6 times a day, or in sustained release form. For most large mammals, the total daily dosage is about 1 mg to about 1000 mg, preferably about 1 mg to about 50 mg. For a 70 kg adult, the total daily dosage is generally about 7 mg to about 350 mg. This dosage regimen may be adjusted to provide the optimal therapeutic response.

[0221] The compounds of the present invention are effective in treating cancer. Without being bound by theory, this is due to the ability of the compounds to modulate the lactate production pathway.

[0222] The compounds of the present invention may be administered with one or more therapeutic agents for cancer selected from the group consisting of PD-1 agents, PD-L1 agents, CTLA-4 agents, IDO1 inhibitors, and anti-cancer vaccines. Alternatively, the compounds may be administered together with cytokine therapy, known chemotherapy or drug therapy, or during radiation therapy. Such combination therapy may be a replacement or a complement to the combination therapy with biguanides discussed above.

[0223] PD-1 agents include, but are not limited to, pembrolizumab and nivolumab.

[0224] PD-L1 agents include, but are not limited to, atezolizumab, avelumab, and durvalumab.

[0225] CTLA-4 agents include, but are not limited to, ipilimumab.

[0226] IDO1 inhibitors include, but are not limited to, epacadostat, navoximod, and BMS-986205.

[0227] Anti-cancer vaccines include, but are not limited to, Hepa-VAC-101 and sipuleucel-T.

[0228] Cytokine therapy includes, but is not limited to, treatment involving administration of IL-2, GM-CSF, IL-12 and / or IL-10.

[0229] Examples of drugs that may be used in the chemotherapy or drug therapy herein include: platinum compounds (e.g., cisplatin, carboplatin, oxaliplatin), alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, nitrogen mustard, thiotepa, melphalan, busulfan, procarbazine, streptomycin, temozolomide, dacarbazine, bendamustine), antitumor antibiotics (e.g., daunorubicin, doxorubicin, idarubicin, epirubicin, mitoxantrone, bleomycin, mitomycin C, plicamycin, , dactinomycin), taxanes (e.g., paclitaxel, cabazitaxel, and docetaxel), antimetabolites (e.g., 5-fluorouracil, cytarabine, pemetrexed, thioguanine, floxuridine, capecitabine, gemcitabine, 6-mercaptopurine, and methotrexate), nucleoside analogues (e.g., fludarabine, clofarabine, cladribine, pentostatin, nelarabine), topoisomerase inhibitors (e.g., topotecan and irinotecan), hypomethylating agents (e.g., azacitidine and decitabine), proteasome inhibitors anti-cancer drugs (e.g., bortezomib), epipodophyllotoxins (e.g., etoposide and teniposide), DNA synthesis inhibitors (e.g., hydroxyurea), vinca alkaloids (e.g., vincristine, vindesine, vinorelbine, and vinblastine), tyrosine kinase inhibitors (e.g., imatinib, dasatinib, nilotinib, sorafenib, sunitinib, bosutinib, ponatinib, erlotinib, gefitinib, afatinib, osimertinib, lapatinib, crizotinib, ceritinib, axitinib, cabozantinib, and lenvatinib), monoclonal antibodies (e.g., bromodiamine ... monoclonal antibodies (e.g., pertuzumab, rituximab, cetuximab, panitumumab, tositumomab, trastuzumab, alemtuzumab, gemtuzumab ozogamicin, bevacizumab, nivolumab, pembrolizumab or other immune checkpoint inhibitor monoclonal antibodies), nitrosoureas (e.g., carmustine, fotemustine, and lomustine), enzymes (e.g., L-asparaginase), biological agents (e.g., interferons and interleukins), hexamethylmelamine, mitotane, angiogenesis inhibitors (e.g., thalidomide, lenalidomide),steroids (e.g., prednisone, dexamethasone, and prednisolone), hormones (e.g., tamoxifen, raloxifene, leuprolide, bicalutamide, granisetron, flutamide), aromatase inhibitors (e.g., letrozole, exemestane, and anastrozole), arsenic trioxide, tretinoin, non-selective cyclooxygenase inhibitors (e.g., nonsteroidal anti-inflammatory drugs, salicylates, aspirin, piroxicam, ibuprofen, indomethacin, naprosyn, diclofenac, tolmetin, ketoprofen, nabumetone, oxaprozin), selective cyclooxygenase-2 (COX-2) inhibitors, or any combination thereof.

[0230] (Abbreviation) The following abbreviations are used herein and throughout this application: Ac Acetyl 9-BBN 9-Borabicyclo[3.3.1]nonane CDCl 3 Deuterated chloroform d double line dba Dibenzylideneacetone dd Double line double line DCE 1,2-dichloroethane DIBAL Diisobutylaluminum hydride DCM Dichloromethane DIAD Diisopropyl azodicarboxylate DIPEA N,N-Diisopropylethylamine DMAP 4-(Dimethylamino)pyridine DMF N,N-Dimethylformamide DMP Dess-Martin oxidation DMSO Dimethyl sulfoxide DMSO-d 6 Deuterated dimethyl sulfoxide dppf 1,1'-bis(diphenylphosphino)ferrocene Et Ethyl EtOAc Ethyl acetate EtOH Ethanol ESI Electrospray Ionization HATU O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HPLC High Performance Liquid Chromatography m multiplet m / z mass-to-charge ratio M mass or mole Me Methyl MeCN Acetonitrile MeOH Methanol MeOD-d 3 Deuterated Methanol MS mass spectrometry NBS N-Bromosuccinimide NMI N-Methylimidazole NMR nuclear magnetic resonance PyBroP Bromotripyrrolidinophosphonium hexafluorophosphate PE Petroleum Ether p-TsCl p-Toluenesulfonyl chloride rt room temperature s single line t triple line TEA Triethylamine TCFH Chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate TFA Trifluoroacetic acid TFAA Trifluoroacetic anhydride THF Tetrahydrofuran Ts Tosil XPhos 2-Dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl

[0231] (General scheme) The compounds of the present invention can be prepared by a combination of methods known in the art, including the procedures described below in Schemes 1 and 2. The following reaction schemes are intended only to illustrate the present invention, and not to limit the present invention. [ka]

[0232] Scheme 1 describes one route for the preparation of compounds of the invention. Amide coupling of a substituted nitroaniline starting material (A-1) with a suitable carboxylic acid A-2, for example with TFCH / NMI as coupling reagent, gives a monoamide intermediate of structure A-3. Reduction of the nitro group in A-3, for example with hydrogen and Pd catalysis, gives an amino intermediate of structure A-4. A second amide coupling in the reaction sequence with a suitable carboxylic acid A-5 converts A-4 to a compound of structure A-6. [ka]

[0233] Scheme 2 shows the specific R 4 The preparation of some of the carboxylic acid intermediates A-5 bearing the hydroxy group is described. The hydroxy-substituted bicyclic heteroaryl starting material (B-1) can be reacted with the appropriate bromide B-2 and, for example, K- 2 CO 3 This can be converted to B-3 via alkylation with .Ester saponification affords the corresponding carboxylic acid intermediate B-4.

[0234] Intermediate 1: N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) [ka]

[0235] Step 1: N-(4-methyl-3-nitrophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1a) To a solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (7.04 g, 39.1 mmol) in MeCN (120 mL) was added 4-methyl-3-nitroaniline (6.00 g, 39.4 mmol), NMI (11.2 g, 137 mmol) and TCFH (19.8 g, 70.4 mmol). The mixture was stirred at room temperature for 16 h. Then, water (120 mL) was added to the mixture. The mixture was filtered and the filter cake was triturated with MeCN / water (1:1) to give the title compound as a yellow solid.

[0236] Step 2: N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) To a solution of N-(4-methyl-3-nitrophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1a) (11.3 g, 36.0 mmol) in MeOH (260 mL) was added Pd / C (1.20 g, 11.3 mmol) and the mixture was heated at room temperature for 2 h. 2 The mixture was stirred under atmosphere overnight. The mixture was then filtered, and the filtrate was concentrated to dryness. The residue was triturated with MeOH to give the title compound as a white solid.

[0237] Intermediate 2: N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) [ka]

[0238] Step 1: N-(4-fluoro-3-nitrophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2a) To a solution of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (6.35 g, 35.3 mmol) and NMI (10.1 g, 123 mmol) in MeCN (150 mL) was added TCFH (17.8 g, 63.5 mmol) and the mixture was stirred at room temperature for 1 h. Then 4-fluoro-3-nitroaniline (5.50 g, 35.3 mmol) was carefully added while cooling with an ice-water bath and the mixture was stirred for 3 h. The mixture was concentrated to dryness and the residue was triturated with MeCN / water (1:1) to give the title compound as a pale yellow solid.

[0239] Step 2: N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2a) (7.80 g, 24.5 mmol) and NH 4 To a mixture of Cl (7.92 g, 147 mmol) in EtOH (120 mL) and water (40.0 mL), iron powder (8.22 g, 147 mmol) was added with N 2 The mixture was added under atmospheric pressure and allowed to stir at 80° C. for 1 h. The mixture was then cooled to rt and filtered. The filtrate was concentrated to dryness and the residue was triturated with MeCN / water (1:1) to give the title compound as a yellow solid.

[0240] Intermediate 2 / 1: N-(3-amino-4-chlorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2 / 1) [ka]

[0241] The title compound was prepared similarly as described for Int2, using 4-chloro-3-nitroaniline in step 1 instead of 4-fluoro-3-nitroaniline.

[0242] Intermediate 3: 5-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylic acid (Int3) [ka]

[0243] Step 1: Ethyl 5-methoxybenzo[b]thiophene-2-carboxylate (Int3a) To a solution of ethyl 2-mercaptoacetate (4.30 mL, 38.9 mmol) in DMSO (20.0 mL), 2 CO 3 (9.00 g, 64.9 mmol) was added at 0° C. and the mixture was stirred at 0° C. for 20 min. A solution of 2-fluoro-5-methoxybenzaldehyde (5.00 g, 32.4 mmol) in DMSO (10.0 mL) was added and the mixture was stirred at 70° C. for 9 h. The mixture was cooled to rt and poured into water (100 mL). The resulting mixture was extracted with DCM (3×100 mL). The combined organic layers were washed with brine and diluted with Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (DCM / PE=2:3) to give the title compound as a green solid.

[0244] Step 2: Ethyl 5-hydroxybenzo[b]thiophene-2-carboxylate (Int3b) To a solution of ethyl 5-methoxybenzo[b]thiophene-2-carboxylate (Int3a) (1.00 g, 4.40 mmol) in DCM (20.0 mL) was added BBr 3 (1M in DCM, 22.0 mL, 21.9 mmol) was added dropwise at -20°C. After the addition, the mixture was slowly warmed to rt and stirred for 2 h. Water (20 mL) was added to the mixture. The resulting mixture was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine and diluted with Na 2 SO 4 It was dried over water, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (EtOAc / PE=1:2) to give the title compound as a pale yellow solid.

[0245] Step 3: Ethyl 5-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylate (Int3c) To a solution of ethyl 5-hydroxybenzo[b]thiophene-2-carboxylate (Int3b) (640 mg, 2.90 mmol) and 1-(3-bromopropyl)pyrrolidine hydrobromide (500 mg, 1.80 mmol) in DMSO (10.0 mL) was added K 2 CO 3 (460 mg, 3.40 mmol) was added and the mixture was stirred at 60° C. overnight. The mixture was then quenched with water (20 mL). The resulting mixture was extracted with DCM (3×10 mL). The combined organic layers were washed with brine and diluted with Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (MeOH / DCM=1:10) to give the title compound as a pale yellow solid.

[0246] Step 4: 5-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylic acid (Int3) To a mixture of ethyl 5-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylate (Int3c) (790 mg, 2.40 mmol) in EtOH (15.0 mL) and water (7.50 mL) was added KOH (324 mg, 5.60 mmol) and the mixture was refluxed for 1 h. The mixture was then cooled to rt and adjusted to pH=5-6 by 1M HCl. The resulting mixture was filtered and the filter cake was triturated with MeCN to give the title compound as an off-white solid.

[0247] Intermediate 4: 6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxylic acid (Int4) [ka]

[0248] Step 1: Ethyl 6-methoxybenzo[b]thiophene-2-carboxylate (Int4a) To a mixture of 2-fluoro-4-methoxybenzaldehyde (10.0 g, 64.9 mmol) and ethyl-2-mercaptoacetate (9.36 g, 77.9 mmol) in DMF (50.0 mL) was added K 2 CO 3 (22.4 g, 162 mmol) was added and the mixture was stirred at room temperature overnight. The mixture was then diluted with water (150 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine and sodium 2 SO 4 It was dried over, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (EtOAc / PE=1:10) to give the title compound as a white solid.

[0249] Step 2: Ethyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int4b) To a solution of ethyl 6-methoxybenzo[b]thiophene-2-carboxylate (Int4b) (10.0 g, 42.4 mmol) in dry DCM (200 mL) was added BBr 3 (1.00M in DCM, 106mL, 106mmol) 2 Dropwise addition was performed under atmospheric pressure at −75° C. After the addition, the mixture was stirred at −75° C. for 2 h and at room temperature for another 16 h. EtOH (30 mL) and saturated aqueous NaHCO 3 (30 mL) was added to the mixture. The resulting mixture was extracted with DCM (3×100 mL) and the combined organic layers were washed with brine and NaSO 4 It was dried over, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (EtOAc / PE=1:6) to give the title compound as a pale yellow solid.

[0250] Step 3: Ethyl 6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxylate (Int4c) Ethyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int4b) (1.20 g, 5.40 mmol) and K 2CO 3 To a mixture of (2.23 g, 16.2 mmol) in MeCN (10.0 mL) was added 2-(bromomethyl)pyridine (1.11 g, 6.50 mmol) and the mixture was stirred at 85° C. overnight. The mixture was cooled to rt and filtered. The filtrate was concentrated to dryness and the residue was purified by column chromatography on silica gel (EtOAc / PE=1:6) to give the title compound as a pale yellow solid.

[0251] Step 4: 6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxylic acid (Int4) To a solution of ethyl 6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxylate (Int4c) (1.20 g, 3.80 mmol) in EtOH (10.0 mL) and water (2.00 mL), LiOH (275 mg, 11.5 mmol) was added and the mixture was stirred at room temperature for 6 h. The mixture was diluted with water (10.0 mL) and the aqueous layer was extracted with DCM (3×30 mL). The aqueous layer was adjusted to pH=5 with 1 M HCl to precipitate a solid. The resulting mixture was filtered and the filter cake was triturated with MeCN to give the title compound as a pale yellow solid.

[0252] Intermediate 5: 6-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylic acid (Int5) [ka]

[0253] Step 1: Ethyl 6-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylate (Int5a) To a mixture of ethyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int4b) (700 mg, 3.20 mmol) and 1-(3-bromopropyl)pyrrolidine hydrobromide (1.03 g, 3.80 mmol) in DMF (10.0 mL) was added K 2 CO 3(1.31 g, 9.50 mmol) was added and the mixture was stirred at 75° C. overnight. The mixture was then cooled to rt, diluted with water (20 mL) and extracted with DCM (3×10 mL). The combined organic layers were washed with water and brine and diluted with Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (MeOH / DCM=1:10) to give the title compound as a pale yellow solid.

[0254] Step 2: 6-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylic acid (Int5) To a mixture of ethyl 6-(3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylate (Int5a) (880 mg, 2.64 mmol) in EtOH (15.0 mL) and water (7.50 mL) was added LiOH (254 mg, 10.6 mmol) and the mixture was stirred at room temperature overnight. The mixture was then adjusted to pH=5-6 by 1M HCl. The resulting mixture was filtered and the filter cake was triturated with MeCN to give the title compound as a pale yellow solid.

[0255] Intermediate 6: 6-(2-(pyrrolidin-1-yl)ethoxy)benzo[b]thiophene-2-carboxylic acid (Int6) [ka]

[0256] Step 1: Ethyl 6-(2-(pyrrolidin-1-yl)ethoxy)benzo[b]thiophene-2-carboxylate (Int6a) To a mixture of ethyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int4b) (2.50 g, 11.3 mmol) and 1-(2-chloroethyl)pyrrolidine (1.80 g, 13.1 mmol) in DMF (25.0 mL) was added K 2 CO 3(4.65 g, 33.8 mmol) was added and the mixture was stirred at 75° C. overnight. The mixture was then cooled to rt, diluted with water (100 mL) and extracted with DCM (3×50 mL). The combined organic layers were washed with water and brine and washed with anhydrous Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (MeOH / DCM=1:10) to give the title compound as a pale yellow solid.

[0257] Step 2: 6-(2-(pyrrolidin-1-yl)ethoxy)benzo[b]thiophene-2-carboxylic acid (Int6) To a mixture of ethyl 6-(2-(pyrrolidin-1-yl)ethoxy)benzo[b]thiophene-2-carboxylate (Int6a) (1.40 g, 4.40 mmol) in EtOH (25.0 mL) and water (5.00 mL) was added LiOH (315 mg, 3.10 mmol) and the mixture was stirred at room temperature overnight. Then EtOH was removed under reduced pressure. The mixture was adjusted to pH=5-6 by 1M HCl. The resulting mixture was filtered and the filter cake was triturated with MeCN to give the title compound as a pale yellow solid.

[0258] Intermediate 7: 6-(2-(pyridin-2-yl)ethoxy)benzo[b]thiophene-2-carboxylic acid (Int7) [ka]

[0259] Step 1: Ethyl 6-(2-(pyridin-2-yl)ethoxy)benzo[b]thiophene-2-carboxylate (Int7a) To a mixture of ethyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int4b) (2.00 g, 9.00 mmol) and 2-(pyridin-2-yl)ethan-1-ol (1.66 g, 13.5 mmol) in THF (40.0 mL) was added PPh 3(7.08 g, 27.0 mmol) and DIAD (5.46 g, 27.0 mmol) were added and the mixture was stirred overnight at 50° C. The mixture was concentrated to dryness and the residue was purified by column chromatography on silica gel (EtOAc / PE=1:5) to give the title compound as a pale yellow solid.

[0260] Step 2: 6-(2-(pyridin-1-yl)ethoxy)benzo[b]thiophene-2-carboxylic acid (Int7) To a solution of ethyl 6-(2-(pyridin-2-yl)ethoxy)benzo[b]thiophene-2-carboxylate (Int7a) (1.10 g, 3.40 mmol) in EtOH (10.0 mL) and water (2.00 mL) was added LiOH (241 mg, 10.1 mmol) and the mixture was stirred at room temperature for 6 h. The mixture was diluted with water (10.0 mL) and the aqueous layer was extracted with DCM (3×30 mL). The aqueous layer was adjusted to pH=5 with 1 M HCl to precipitate a solid. The resulting mixture was filtered and the filter cake was triturated with MeCN to give the title compound as a pale yellow solid.

[0261] Intermediate 8: 6-Methoxybenzo[d]thiazole-2-carboxylic acid (Int8) [ka]

[0262] To a solution of 6-methoxybenzo[d]thiazole-2-carbonitrile (30.0 mg, 0.158 mmol) in THF (1.60 mL) was added 2M NaOH (0.118 mL, 0.237 mmol). The mixture was stirred at room temperature for 48 h. The reaction mixture was diluted with EtOAc (20 mL) and water (10 mL). The organic layer was separated. The aqueous layer was acidified to pH ∼1 with 1M HCl and then extracted with EtOAc (2 × 15 mL). The organic layer was diluted with MgSO 4 It was dried over water, filtered and concentrated to give the title compound as a yellow solid.

[0263] Intermediate 9: 6-((4-ethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int9) [ka]

[0264] Step 1: Ethyl thieno[2,3-b]pyridine-2-carboxylate (Int9a) To a solution of 2-chloronicotinaldehyde (50.0 g, 354 mmol) in DMF (400 mL) was added TEA (74.1 g, 732 mmol) and ethyl 2-mercaptoacetate (42.5 g, 354 mmol). The mixture was stirred at 100° C. for 4 h under nitrogen atmosphere. The mixture was diluted with water (1.5 L) and extracted with EtOAc (3×1000 mL). The combined organic layers were washed with water and brine and washed with anhydrous Na 2 SO 4 It was dried over, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica (PE / EtOAc=8:1) to give the title compound.

[0265] Step 2: 2-(ethoxycarbonyl)thieno[2,3-b]pyridine 7-oxide (Int9b) To a solution of ethyl thieno[2,3-b]pyridine-2-carboxylate (Int9a) (52.8 g, 255.0 mmol) in acetic acid (400 mL) was added H 2 O 2 (100 mL, 30% in water) was added portionwise. The resulting mixture was stirred at 60° C. overnight. The mixture was stirred at 0° C. for 1 h. A precipitate formed during the reaction. The solid was collected by filtration and dried to give the title compound.

[0266] Step 3: Ethyl 6-hydroxythieno[2,3-b]pyridine-2-carboxylate (Int9c) To a solution of 2-(ethoxycarbonyl)thieno[2,3-b]pyridine 7-oxide (Int9b) (15.0 g, 67.2 mmol) in DMF (150 mL) was added TFAA (28.0 g, 134 mmol) dropwise at 0° C. The mixture was stirred at 0° C. for 3 h. The mixture was poured into water (300 mL) and stirred for 5 min. A precipitate formed. The solid was collected by filtration and dried to give the title compound as a yellow solid, which was used in the next step without further purification.

[0267] Step 4: Ethyl 6-chlorothieno[2,3-b]pyridine-2-carboxylate (Int9d) To a solution of ethyl 6-hydroxythieno[2,3-b]pyridine-2-carboxylate (Int9c) (14.8 g, 66.3 mmol) in DMF (150 mL) was added POCl 3 (50.7 g, 332 mmol) was added and the mixture was stirred at 100° C. overnight. The mixture was cooled to rt and the solvent was removed under reduced pressure. The residue was diluted with water (200 mL) and saturated aqueous NaHCO 3 The pH was adjusted to 8 with 50 mL of EtOAc. The mixture was extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (200 mL) and anhydrous Na 2 SO 4 It was dried over water, filtered and concentrated to dryness. The crude product was purified by column chromatography (PE / EtOAc=8:1) to give the title compound.

[0268] Step 5: Ethyl 6-methylthieno[2,3-b]pyridine-2-carboxylate (Int9e) Ethyl 6-chlorothieno[2,3-b]pyridine-2-carboxylate (Int9d) (5.0 g, 20.7 mmol), 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (25 g, 200 mmol), Pd(PPh 3 ) 4 (2.3g, 2.0mmol), K 2 CO 3 A solution of (8.4 g, 62.1 mmol) in dioxane (100 mL) was 2The mixture was stirred under atmosphere at 100° C. for 14 h. The mixture was cooled to rt and filtered. The filtrate was concentrated to dryness. The residue was purified by column chromatography (PE / EtOAc=8:1) to give the title compound.

[0269] Step 6: Ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate (Int9f) Ethyl 6-methylthieno[2,3-b]pyridine-2-carboxylate (Int9e) (3.8 g, 17.2 mmol), AIBN (281 mg, 1.72 mmol) and NBS (3.04 g, 17.2 mmol) were dissolved in CCl 4 The solution in (50 mL) was stirred at 80° C. for 14 h. The mixture was cooled to rt and concentrated to dryness. The residue was purified by column chromatography (PE / EtOAc=6:1) to give the title compound.

[0270] Step 7: Ethyl 6-((4-ethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int9g) A solution of ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate (Int9f) (4.6 g, 15.3 mmol) in acetonitrile (100 mL) was added with 1-ethylpiperazine (2.6 g, 23 mmol) and K 2 CO 3 (6.3 g, 45.9 mmol) was added. The mixture was stirred at 70° C. overnight. The mixture was diluted with water (500 mL) and extracted with EtOAc (3×200 mL). The combined organic layers were washed with brine (200 mL) and diluted with anhydrous Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0271] Step 8: 6-((4-Ethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int9) Ethyl 6-((4-ethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int9g) (4.1 g, 12.3 mmol) in MeOH / H2 To a solution in 2H2O (5:1, 50 mL) was added NaOH (984 mg, 24.6 mmol) and the mixture was stirred at room temperature overnight. The mixture was diluted with water and adjusted to pH = 5 using 1 M HCl. The mixture was extracted with EtOAc (3 x 200 mL). The aqueous phase was concentrated to dryness and the residue was purified by reverse phase column chromatography (gradient acetonitrile / H2O 2 O=5%-80%) to give the title compound.

[0272] Intermediate 10: 6-((4-ethyl-3,3-dimethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10) [ka]

[0273] Step 1: Ethyl 6-((4-ethyl-3,3-dimethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int10a) To a mixture of ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate (Int9f) (50.0 mg, 0.167 mmol) and cesium carbonate (162.8 mg, 0.500 mmol) in dry THF (2 mL) was added 1-ethyl-2,2-dimethylpiperazine (23.7 mg, 0.167 mmol) and the mixture was stirred overnight at 50° C. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0274] Step 2: 6-((4-ethyl-3,3-dimethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10) MeOH and H 2To ethyl 6-((4-ethyl-3,3-dimethylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int10a) (55.1 mg, 0.152 mmol) dissolved in a mixture of 2H2O (3:1, 4 mL) was added sodium hydroxide (2 M aqueous solution) (1 mL) and the mixture was stirred at 50° C. for 1 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white powder.

[0275] Intermediate 10 / 1: 6-((3,3-difluoroazetidin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10 / 1) [ka]

[0276] The title compound was prepared similarly as described for Int10, using 3,3-difluoroazetidine hydrochloride in step 1 instead of 1-ethyl-2,2-dimethylpiperazine.

[0277] Intermediate 10 / 2: 6-(((1-ethylpiperidin-4-yl)amino)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10 / 2) [ka]

[0278] The title compound was prepared similarly as described for Int10, using 1-ethylpiperidin-4-amine in step 1 instead of 1-ethyl-2,2-dimethylpiperazine.

[0279] Intermediate 10 / 3: 6-((diethylamino)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10 / 3) [ka]

[0280] The title compound was prepared similarly as described for Int10, using diethylamine in step 1 instead of 1-ethyl-2,2-dimethylpiperazine.

[0281] Intermediate 10 / 4: 6-(piperazin-1-ylmethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10 / 4) [ka]

[0282] The title compound was prepared similarly as described for Intermediate 10, using 1-(piperazin-1-yl)ethan-1-one in step 1 instead of 1-ethyl-2,2-dimethylpiperazine.

[0283] Intermediate 11: 6-((4-ethylpiperazin-1-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int11) [ka]

[0284] Step 1: Methyl 6-methylbenzo[b]thiophene-2-carboxylate (Int11a) To a solution of 6-methylbenzo[b]thiophene-2-carboxylic acid (600.0 mg, 3.121 mmol) in dry MeOH (20.0 mL) was added SOCl 2 (0.250 mL, 3.433 mmol) was added dropwise at 0° C. The mixture was allowed to warm to room temperature and then refluxed for 18 h. The mixture was concentrated to dryness and the crude product was used in the next step without further purification.

[0285] Step 2: Methyl 6-(bromomethyl)benzo[b]thiophene-2-carboxylate (Int11b) To a solution of methyl 6-methylbenzo[b]thiophene-2-carboxylate (Int11a) (644 mg, 3.121 mmol) in dry MeCN (15.6 mL) was added NBS (611 mg, 3.433 mmol) and benzoyl peroxide (151 mg, 0.468 mmol) at room temperature. The mixture was refluxed for 18 h. The mixture was cooled to rt and diluted with EtOAc (50 mL). The organic layer was extracted with NaHCO 3 (20 mL), water (20 mL) and brine (20 mL). The organic layer was washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel using a gradient of 0-4% EtOAc in cyclohexane to give the title compound.

[0286] Step 3: Methyl 6-((4-ethylpiperazin-1-yl)methyl)benzo[b]thiophene-2-carboxylate (Int11c) To a solution of methyl 6-(bromomethyl)benzo[b]thiophene-2-carboxylate (Int11b) (522 mg, 0.915 mmol) in dry THF (4.6 mL) was added Cs 2 CO 3 (328 mg, 1.007 mmol) and 1-ethylpiperazine (0.116 mL, 0.915 mmol) were added. The mixture was stirred at 60° C. for 3 h. The mixture was taken on directly to the next step.

[0287] Step 4: 6-((4-ethylpiperazin-1-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int11) Methyl 6-((4-ethylpiperazin-1-yl)methyl)benzo[b]thiophene-2-carboxylate (Int11c) (mixture from step 3) was added to water (1 mL) and LiOH H 2 O (115 mg, 2.746 mmol) was added. The mixture was stirred at room temperature for 18 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white solid.

[0288] Intermediate 11 / 1: 6-((4-(2,2,2-trifluoroethyl)piperazin-1-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int11 / 1) [ka]

[0289] The title compound was prepared similarly as described for Intermediate 11, using 1-(2,2,2-trifluoroethyl)piperazine in step 3 instead of 1-ethylpiperazine.

[0290] Intermediate 11 / 2: 6-((4-methylpiperazin-1-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int11 / 2) [ka]

[0291] The title compound was prepared similarly as described for Intermediate 11, using 1-methylpiperazine in step 3 instead of 1-ethylpiperazine.

[0292] Intermediate 11 / 3: 6-(morpholinomethyl)benzo[b]thiophene-2-carboxylic acid (Int11 / 3) [ka]

[0293] The title compound was prepared similarly as described for Intermediate 11, using morpholine in step 3 instead of 1-ethylpiperazine.

[0294] Intermediate 11 / 4: 6-((4-(tert-butoxycarbonyl)-1,4-diazepan-1-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int11 / 4) [ka]

[0295] The title compound was prepared similarly as described for Intermediate 11, using tert-butyl 1,4-diazepane-1-carboxylate in step 3 instead of 1-ethylpiperazine.

[0296] Intermediate 12: 6-(3-(3-methoxyazetidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylic acid (Int12) [ka]

[0297] Step 1: Methyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int12a) To a mixture of methyl 6-methoxybenzo[b]thiophene-2-carboxylate (200 mg, 0.90 mmol) in dry DCM (5 mL), boron tribromide (1.00 M in DCM, 1.8 mL, 1.80 mmol) was added with N 2 Dropwise addition was performed under atmospheric pressure at −75° C. After the addition, the mixture was stirred at −75° C. for 2 h and at room temperature for another 16 h. EtOH (20 mL) and saturated aqueous NaHCO 3 (20 mL) was added to the mixture. The mixture was extracted with DCM (3×50 mL) and the combined organic layers were washed with brine and Na 2 SO 4 It was dried over, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (EtOAc / PE=1:6) to give the title compound as a pale yellow solid.

[0298] Step 2: Methyl 6-(3-(3-methoxyazetidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylate (Int12b) Methyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int12a) (50 mg, 0.24 mmol), 1-bromo-3-chloropropane (28 μL, 0.29 mmol) and cesium carbonate (172 mg, 0.53 mmol) were dissolved in dry DMF (2 mL) and the mixture was stirred overnight at 50° C. 3-Methoxyazetidine hydrochloride (35.6 mg, 0.29 mmol) was added and the mixture was stirred at 50° C. for 2 h. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0299] Step 3: 6-(3-(3-methoxyazetidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylic acid (Int12) Methyl 6-(3-(3-methoxyazetidin-1-yl)propoxy)benzo[b]thiophene-2-carboxylate (Int12b) (80 mg, 0.24 mmol) in THF / H 2 To the mixture in O (3:1) was added lithium hydroxide monohydrate (50 mg, 1.20 mmol) and the mixture was stirred at room temperature overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white powder.

[0300] Intermediate 13: 6-(2-acetamidoethoxy)benzo[b]thiophene-2-carboxylic acid (Int13) [ka]

[0301] Step 1: Methyl 6-(2-((tert-butoxycarbonyl)amino)ethoxy)benzo[b]thiophene-2-carboxylate (Int13a) Methyl 6-hydroxybenzo[b]thiophene-2-carboxylate (Int12a) (50.0 mg, 0.240 mmol) and Cs 2 CO 3To a solution of (93.9 mg, 0.288 mmol) in DMF (2.4 mL) was added tert-butyl(2-bromoethyl)carbamate (83.9 mg, 0.288 mmol). The mixture was stirred at 60 °C overnight. Water was added (10 mL) and the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with MgSO 4 Drying on ice, filtration and concentration to dryness afforded the title compound as a yellow oil which was used in the next step without further purification.

[0302] Step 2: Methyl 6-(2-aminoethoxy)benzo[b]thiophene-2-carboxylate (Int13b) TFA (1.4 mL) was added to a solution of methyl 6-(2-((tert-butoxycarbonyl)amino)ethoxy)benzo[b]thiophene-2-carboxylate (Int13a) (100 mg, 0.285 mmol) in DCM (2.8 mL) at 0° C. The mixture was stirred at room temperature overnight. Saturated aqueous NaHCO 3 (2 mL) was added and the mixture was extracted with EtOAc (3×10 mL). The combined organic layers were diluted with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0303] Step 3: Methyl 6-(2-acetamidoethoxy)benzo[b]thiophene-2-carboxylate (Int13c) Methyl 6-(2-aminoethoxy)benzo[b]thiophene-2-carboxylate (Int13b) (20.0 mg, 0.080 mmol) was dissolved in THF (1.7 mL). Triethylamine (0.022 mL, 0.159 mmol) was added, followed by dropwise addition of acetyl chloride (0.006 mL, 0.088 mmol). The mixture was stirred at rt overnight. Water (5 mL) was added and the mixture was extracted with EtOAc (3×10 mL). The combined organic layers were washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0304] Step 4: 6-(2-acetamidoethoxy)benzo[b]thiophene-2-carboxylic acid (Int13) A solution of methyl 6-(2-acetamidoethoxy)benzo[b]thiophene-2-carboxylate (Int13c) (24.0 mg, 0.082 mmol) in THF (0.8 mL) and water (0.8 mL) was added with LiOH H 2 O (17.5 mg, 0.417 mmol) was added and the mixture was stirred at room temperature overnight. 1M HCl was added to adjust the pH to 5. The mixture was extracted with EtOAc (3×10 mL). The combined organic layers were washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0305] Intermediate 14: Methyl 6-chlorothieno[2,3-b]pyridine-2-carboxylate (Int14) [ka]

[0306] The title compound was prepared similarly as described for Intermediate 9d, steps 1-4, using methyl 2-mercaptoacetate in step 1 instead of ethyl 2-mercaptoacetate.

[0307] Intermediate 15: 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15) [ka]

[0308] Step 1: Methyl 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylate (Int15a) To a solution of 2-(pyrrolidin-1-yl)ethan-1-ol (0.54 g, 4.68 mmol) in THF (50 mL) was added NaH (60% dispersion in mineral oil, 0.374 g, 15.6 mmol) in portions at 0° C. The mixture was stirred at 0° C. for 5 min, then warmed to rt and stirred at that temperature for 1 h. Methyl 6-chlorothieno[2,3-b]pyridine-2-carboxylate (Int14) (1.0 g, 4.4 mmol) was added and the mixture was stirred at 75° C. for 20 min. The mixture was cooled to 0° C. and diluted with H 2 The mixture was quenched with 20 mL of O and extracted with EtOAc (3×30 mL). The combined organic layers were washed with water and brine, and washed with anhydrous Na 2 SO 4 It was dried over hexane, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica (DCM / MeOH=20:1) to give the title compound.

[0309] Step 2: 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15) To a solution of methyl 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylate (Int15a) (850 mg, 2.77 mmol) in EtOH (15 mL) was added a solution of NaOH (359 mg, 5.97 mmol in 5 mL water). The mixture was stirred at 70° C. for 1 h. The mixture was diluted with water and adjusted to pH=6 using 1M HCl. The mixture was filtered and the filtrate was concentrated to dryness. The residue was purified by reverse phase column chromatography (solvent A:H 2 0, solvent B: MeCN) to give the title compound.

[0310] Intermediate 15 / 1: 6-(3-(pyrrolidin-1-yl)propoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15 / 1) [ka]

[0311] The title compound was prepared similarly as described for Intermediate 15, using 3-(pyrrolidin-1-yl)propan-1-ol in step 1 instead of 2-(pyrrolidin-1-yl)ethan-1-ol.

[0312] Intermediate 16: 6-((pyridin-2-yloxy)methyl)benzo[b]thiophene-2-carboxylic acid (Int16) [ka]

[0313] Step 1: 6-(Hydroxymethyl)benzo[b]thiophene-2-carboxylic acid (Int16a) To a mixture of methyl 6-(bromomethyl)benzo[b]thiophene-2-carboxylate (Int11b) (3.0 g, 10.5 mmol) in dioxane (100 mL) was added H 2 K in O (100 mL) 2 CO 3 (5.8 g, 42 mmol) was added. The mixture was stirred at 100° C. for 1 h. Aqueous HCl was added to the mixture to adjust the pH=2. The aqueous layer was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (100 mL) and diluted with Na 2 SO 4 It was dried over water and concentrated to dryness to give the title compound as a yellow solid.

[0314] Step 2: 6-((pyridin-2-yloxy)methyl)benzo[b]thiophene-2-carboxylic acid (Int16) To a mixture of 6-(hydroxymethyl)benzo[b]thiophene-2-carboxylic acid (Int16a) (2.2 g, 10.56 mmol), 2-chloropyridine (3.56 g, 31.68 mmol) and 18-crown-6 (0.44 g, 1.66 mmol) in toluene (150 mL) was added t-BuOK (11.86 g, 105.6 mmol). The mixture was stirred at 120° C. for 6 h. The mixture was then cooled to 10° C. for 2 h. 2The aqueous layer was adjusted to pH = 2 with aqueous HCl and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with Na 2 SO 4 It was dried on a stover, filtered and concentrated to dryness to give the title compound as a white solid which was used in the next step without further purification.

[0315] Intermediate 17: 6-(((3aR,6aS)-5-ethylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int17) [ka]

[0316] Step 1: 6-(((3aR,6aS)-Hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate ethyl (Int17a)

[0317] A mixture of ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate (Int9f) (30 mg, 0.10 mmol), DCM (0.5 mL), triethylamine (11.1 mg, 0.11 mmol) and tert-butyl (3aR,6aS)-3a,6a-dimethylhexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (22.3 mg, 0.11 mmol) was stirred at room temperature overnight. TFA (0.25 mL) was added and the mixture was stirred at room temperature overnight. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0318] Step 2: 6-(((3aR,6aS)-5-Ethylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int17) To a solution of ethyl 6-(((3aR,6aS)-3a,6a-dimethylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int17a) (25 mg, 0.075 mmol) in dry THF was added Cs 2 CO 3 (24 mg, 0.075 mmol) and iodoethane (12 mg, 0.075 mmol) were added. The mixture was stirred at room temperature for 2 h. Aqueous NaOH was added (2 M, 200 μL) and the mixture was stirred at room temperature for 2 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound.

[0319] Intermediate 18: Methyl 6-hydroxythieno[2,3-b]pyridine-2-carboxylate (Int18) [ka]

[0320] The title compound was prepared similarly as described for Intermediate 9c steps 1-3, using methyl 2-mercaptoacetate in step 1 instead of ethyl 2-mercaptoacetate.

[0321] Intermediate 19: 6-(2-(4-ethylpiperazin-1-yl)ethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int19) [ka]

[0322] Step 1: Methyl 6-bromothieno[2,3-b]pyridine-2-carboxylate (Int19a) Methyl 6-hydroxythieno[2,3-b]pyridine-2-carboxylate (Int18) (15.7 g, 75 mmol) and POBr 3( A mixture of 96.2 mL, 0.96 mol) in DMF (150 mL) was stirred at 120° C. for 16 h. The mixture was concentrated and diluted with DCM (300 mL). The organic layer was washed with saturated aqueous NaHCO3 (2×80 mL) and brine (2×100 mL), and then washed with anhydrous Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness. The residue was purified by silica gel column chromatography (DCM / MeOH=20:1) to give the title compound.

[0323] Step 2: (E)-Methyl 6-(2-ethoxyvinyl)thieno[2,3-b]pyridine-2-carboxylate (Int19b) To a mixture of methyl 6-bromothieno[2,3-b]pyridine-2-carboxylate (Int19a) (2.1 g, 7.7 mmol) and KF (893 mg, 15.37 mmol) in 1,4-dioxane (40 mL) and water (8 mL) was added tricyclohexylphosphine (418 mg, 1.49 mmol), (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.75 g, 8.8 mmol) and Pd 2 (dba) 3 (673 mg, 0.74 mmol) was added. The mixture was stirred at 100° C. under nitrogen atmosphere for 16 h. The mixture was cooled to rt, diluted with water and extracted with EtOAc (3×100 mL). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The residue was purified by silica gel chromatography (PE / EtOAc=1:1) to give the title compound.

[0324] Step 3: Methyl 6-(2-oxoethyl)thieno[2,3-b]pyridine-2-carboxylate (Int19c) To a solution of (E)-methyl 6-(2-ethoxyvinyl)thieno[2,3-b]pyridine-2-carboxylate (Int19b) (0.6 g, 2.28 mmol) in DCM (10 mL) was added concentrated HCl (6 mL) and the mixture was stirred at room temperature for 16 h. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0325] Step 4: Methyl 6-(2-(4-ethylpiperazin-1-yl)ethyl)thieno[2,3-b]pyridine-2-carboxylate (Int19d) To a solution of methyl 6-(2-oxoethyl)thieno[2,3-b]pyridine-2-carboxylate (Int19c) (400 mg, 1.7 mmol), 1-ethylpiperazine (213.5 mg, 1.86 mmol) and sodium triacetoxyborohydride (1.08 g, 5.1 mmol) in DCE (50 mL) was added AcOH (0.2 mL). The mixture was stirred at room temperature for 16 h. The mixture was diluted with water (80 mL) and extracted with DCM (3×100 mL). The combined organic layers were washed with water and brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The residue was purified by silica gel chromatography (EtOAc / PE=1:5) to give the title compound.

[0326] Step 5: 6-(2-(4-ethylpiperazin-1-yl)ethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int19) To a solution of methyl 6-(2-(4-ethylpiperazin-1-yl)ethyl)thieno[2,3-b]pyridine-2-carboxylate (Int19d) (350 mg, 1.05 mmol) in MeOH (10 mL) and water (2 mL) was added LiOH (66 mg, 2.76 mmol). The mixture was stirred at 60° C. for 2 h. The mixture was adjusted to pH=5 using 1M HCl and extracted with EtOAc (2×20 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0327] Intermediate 20: 6-((1-ethylpiperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int20) [ka]

[0328] Step 1: tert-Butyl 4-(((1s,5s)-9-borabicyclo[3.3.1]nonan-9-yl)methyl)piperidine-1-carboxylate (Int20a) To a solution of tert-butyl 4-methylenepiperidine-1-carboxylate (4.0 g, 0.020 mol) in THF (60 mL) was added 9-BBN (4.9 g, 40 mmol) and the mixture was refluxed for 1 h. The mixture was cooled to rt and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0329] Step 2: Methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int20b) tert-Butyl 4-(((1s,5s)-9-borabicyclo[3.3.1]nonan-9-yl)methyl)piperidine-1-carboxylate (Int20a) (5.0 g, crude) in DMF (40 mL) and H 2 To a solution of Pd(dppf)Cl in 20O (4 mL), 2 (1.6 g, 2.0 mmol), methyl 6-bromothieno[2,3-b]pyridine-2-carboxylate (Int19a) (6.7 g, 0.025 mol) and K 2 CO 3 (4.0 g, 0.029 mol) was added. The mixture was stirred at 60° C. for 16 h under nitrogen atmosphere. The mixture was concentrated under reduced pressure, extracted with EtOAc, and 2 Wash with O and brine, and then with anhydrous Na 2 SO 4 It was dried over water, filtered and concentrated to dryness. The residue was purified by silica gel chromatography (PE / EtOAc=1:1) to give the title compound.

[0330] Step 3: Methyl 6-(piperidin-4-ylmethyl)thieno[2,3-b]pyridine-2-carboxylate (Int20c) To a solution of methyl 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int20b) (2.6 g, 6.7 mmol) in DCM (20 mL) was added TFA (10 mL). The mixture was stirred at room temperature for 1 h. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0331] Step 4: Methyl 6-((1-ethylpiperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int20d) To a solution of methyl 6-(piperidin-4-ylmethyl)thieno[2,3-b]pyridine-2-carboxylate (Int20c) (3 g, crude) in DCE (50 mL) was added acetaldehyde (792 mg, 18 mmol), TEA (2.4 g, 24 mmol). Sodium triacetoxyborohydride (5.0 g, 24 mmol) was added and the mixture was stirred at room temperature for 16 h. Water (50 mL) was added and the mixture was extracted with DCM (2×200 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The residue was purified by silica gel chromatography (DCM / MeOH=20:1) to give the title compound.

[0332] Step 5: 6-((1-ethylpiperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int20) To a solution of methyl 6-((1-ethylpiperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int20d) (400 mg, 1.25 mmol) in MeOH (16 mL) and water (4 mL) was added LiOH (52 mg, 2.2 mmol). The mixture was stirred at 60° C. for 1 h. The pH was adjusted to pH=5 by 1 M HCl and the mixture was extracted with EtOAc (2×20 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0333] Intermediate 21: 6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxylic acid (Int21) [ka]

[0334] Step 1: Ethyl 2-bromoimidazo[2,1-b]thiazole-6-carboxylate (Int21a) To a solution of 2-amino-5-bromothiazole monohydrobromide (20 g, 77 mmol) in ethanol (300 mL) was added ethyl bromopyruvate (19 mL, 154 mmol). The mixture was stirred at reflux overnight. The mixture was concentrated. Water (300 mL) was added to the residue and the aqueous layer was extracted with EtOAc (300 mL). The organic layer was washed with brine (200 mL) and diluted with Na 2 SO 4 It was dried over water, filtered and concentrated to dryness. The residue was purified by flash column chromatography on silica gel (PE / EtOAc=1:1) to give the title compound as a brown solid.

[0335] Step 2: (2-Bromoimidazo[2,1-b]thiazol-6-yl)methanol (Int21b) To a solution of ethyl 2-bromoimidazo[2,1-b]thiazole-6-carboxylate (Int21a) (11.5 g, 42.0 mmol) was added DIBAL-H (42 mL, 1 M in hexane) dropwise under argon at 0° C. over 0.5 h. The mixture was allowed to warm to rt and stirred for 3 h. Sodium sulfate decahydrate (27 g, 83.8 mmol) was added in batches under ice bath cooling. The suspension was filtered through Celite® using MeOH to rinse the Celite® pad. The filtrate was concentrated to dryness and the residue was purified by column chromatography on silica gel (DCM / MeOH=10:1) to give the title compound as a pale yellow solid.

[0336] Step 3: 2-Bromo-6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole (Int21c) To a stirred solution of (2-bromoimidazo[2,1-b]thiazol-6-yl)methanol (Int21b) (1.6 g, 6.9 mmol) and triethylamine (1.9 mL, 13.6 mmol) in dry DCM (20 mL) was added methanesulfonyl chloride (0.8 mL, 10.3 mmol). The mixture was stirred for 0.5 h. The mixture was concentrated and dissolved in dry DCM (20 mL). Triethylamine (1.9 mL, 13.6 mmol) was added followed by 1-ethylpiperazine (0.96 mL, 7.6 mmol). The mixture was stirred at rt for 2 h. The mixture was washed with water (40 mL) and brine (40 mL). The organic layer was extracted with Na 2 SO 4 It was dried over water, filtered and concentrated to dryness. The residue was purified by flash chromatography (DCM / MeOH=10:1) to give the title compound as a pale yellow solid.

[0337] Step 4: Methyl 6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxylate (Int21d) To a solution of 2-bromo-6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole (Int21c) (1.1 g, 3.3 mmol) and triethylamine (1.9 mL, 13.6 mmol) in MeOH (20 mL) was added Pd(OAc) 2 (226 mg, 1.0 mmol) and XPhos (481 mg, 1.0 mmol) were added under argon protection. The mixture was saturated with CO gas and stirred at 60° C. overnight. The mixture was filtered through a pad of Celite®. The filtrate was concentrated to dryness to give the title compound as an orange solid.

[0338] Step 5: 6-((4-Ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxylic acid (Int21) To a solution of methyl 6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxylate (Int21d) (1.0 g, 3.2 mmol) in dry THF (10 mL) was added aqueous LiOH (6.4 mL, 1 M). The mixture was stirred at reflux for 3 h. The mixture was concentrated and aqueous HCl (1 M) was added with stirring until pH=2. The mixture was concentrated to dryness. DCM was added to the residue and the suspension was stirred for 1 h. The mixture was filtered and the filtrate was concentrated to dryness to give the title compound as an orange solid.

[0339] Intermediate 22: 5-(2-(pyrrolidin-1-yl)ethoxy)-1H-indole-2-carboxylic acid (Int22) [ka]

[0340] Step 1: Methyl 5-hydroxy-1H-indole-2-carboxylate (Int22a) A mixture of 5-hydroxy-1H-indole-2-carboxylic acid (500 mg, 2.82 mmol) in methanol (28 mL) was cooled to 0° C. and thionyl chloride (369 mg, 3.11 mmol) was added. The mixture was stirred at 50° C. overnight. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0341] Step 2: Methyl 5-((tert-butyldimethylsilyl)oxy)-1H-indole-2-carboxylate (Int22b) To a mixture of methyl 5-hydroxy-1H-indole-2-carboxylate (Int22a) (540 mg, 2.82 mmol), imidazole (385 mg, 5.65 mmol) in DMF (28 mL) was added dropwise a solution of tert-butylchlorodimethylsilane (511 mg, 3.39 mmol) in DMF. The mixture was stirred at rt overnight. Imidazole (192 mg, 2.82 mmol) and tert-butylchlorodimethylsilane (255 mg, 1.69 mmol) were added and the mixture was stirred at room temperature for 1 h. The mixture was concentrated to dryness. The residue was purified by column chromatography on silica gel (gradient EtOAc / cyclohexane) to give the title compound.

[0342] Step 3: 1-(tert-butyl)2-methyl 5-((tert-butyldimethylsilyl)oxy)-1H-indole-1,2-dicarboxylate (Int22c) A mixture of methyl 5-((tert-butyldimethylsilyl)oxy)-1H-indole-2-carboxylate (Int22b) (561 mg, 1.837 mmol) and NaH (132 mg, 24 mmol) in DMF (18 mL) was stirred at room temperature for 30 min. Di-tert-butyl dicarbonate (521 mg, 2.39 mmol) was added and the mixture was stirred at room temperature overnight. The mixture was concentrated to dryness and the residue was purified by column chromatography on silica gel (gradient EtOAc / cyclohexane) to give the title compound.

[0343] Step 4: 5-Hydroxy-1H-indole-1,2-dicarboxylate 1-(tert-butyl)2-methyl (Int22d) To a mixture of 1-(tert-butyl)2-methyl 5-((tert-butyldimethylsilyl)oxy)-1H-indole-1,2-dicarboxylate (Int22c) (376 mg, 0.927 mmol) in dry THF (10 L) was added dropwise a solution of TBAF in THF (1 M, 0.927 mL, 0.927 mmol) at 0° C. The mixture was stirred at room temperature for 20 min. The mixture was concentrated to dryness and the residue was purified by column chromatography on silica gel (gradient EtOAc / cyclohexane) to give the title compound.

[0344] Step 5: 1-(tert-butyl)2-methyl 5-(2-(pyrrolidin-1-yl)ethoxy)-1H-indole-1,2-dicarboxylate (Int22e) 1-(tert-butyl)2-methyl 5-hydroxy-1H-indole-1,2-dicarboxylate (Int22d) (50 mg, 0.172 mmol) was dissolved in dry DMF. NaH was added at 0° C. and the mixture was stirred at room temperature for 20 min. A solution of 1-(2-chloroethyl)pyrrolidine hydrochloride (29.2 mg, 0.172 mmol) in dry DMF (0.7 mL) was added dropwise and the mixture was stirred at 50° C. overnight. The mixture was concentrated and water was added to the residue (10 mL). The suspension was extracted with EtOAc (3×10 mL). The combined organic layers were washed with anhydrous MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0345] Step 6: Methyl 5-(2-(pyrrolidin-1-yl)ethoxy)-1H-indole-2-carboxylate (Int22f) TFA (600 μL) was added to a solution of 1-(tert-butyl)2-methyl 5-(2-(pyrrolidin-1-yl)ethoxy)-1H-indole-1,2-dicarboxylate (In22e) (47.0 mg, 0.121 mmol) in DCM (1.2 mL) at 0° C. The mixture was stirred at room temperature for 1 h. Water was added and the mixture was extracted with DCM. The combined organic layers were washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0346] Step 7: 5-(2-(pyrrolidin-1-yl)ethoxy)-1H-indole-2-carboxylic acid (Int22) LiOH H 2O (11.0 mg, 0.253 mmol) was added to a mixture of methyl 5-(2-(pyrrolidin-1-yl)ethoxy)-1H-indole-2-carboxylate (Int22f) (11.0 mg, 0.038 mmol) in a mixture of water and THF (1:1, 0.8 mL). The mixture was stirred at rt overnight. The mixture was neutralized with 1M HCl and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0347] Intermediate 23: 6-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23) [ka]

[0348] Step 1: Ethyl 6-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int23a) To a mixture of ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate (Int9f) (50.0 mg, 0.167 mmol) and cesium carbonate (162.8 mg, 0.500 mmol) in dry THF (2 mL) was added 1-tert-butyl piperazine-1-carboxylate (31.0 mg, 0.167 mmol) and the mixture was stirred overnight at 50° C. The mixture was concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0349] Step 2: 6-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23) Ethyl 6-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylate (Int23a) (60 mg, 0.148 mmol) in THF and H 2To the mixture in O (3:1, 4 mL) was added lithium hydroxide monohydrate (10.5 mg, 0.222 mmol) and the mixture was stirred at room temperature overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white powder.

[0350] Intermediate 23 / 1: 6-(((1R,4R)-5-(tert-butoxycarbonyl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23 / 1) [ka]

[0351] The title compound was prepared similarly as described for Int23, using (1R,4R)-tert-butyl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate in step 1 instead of 1-tert-butyl piperazine-1-carboxylate.

[0352] Intermediate 23 / 2: 6-(((1S,4S)-5-(tert-butoxycarbonyl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23 / 2) [ka]

[0353] The title compound was prepared similarly as described for Int23, using (1S,4S)-tert-butyl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate in step 1 instead of 1-tert-butyl piperazine-1-carboxylate.

[0354] Intermediate 23 / 3: 6-((4-(tert-butoxycarbonyl)-1,4-diazepan-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23 / 3) [ka]

[0355] The title compound was prepared similarly as described for Int23, using tert-butyl 1,4-diazepane-1-carboxylate in step 1 instead of 1-tert-butyl piperazine-1-carboxylate.

[0356] Intermediate 23 / 4: 6-(((3aR,6aS)-5-(tert-butoxycarbonyl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int23 / 4) [ka]

[0357] The title compound was prepared similarly as described for Int23, using methyl 6-(bromomethyl)benzo[b]thiophene-2-carboxylate (Int11b) instead of ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate and tert-butyl (3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate in step 1 instead of 1-tert-butyl piperazine-1-carboxylate.

[0358] Intermediate 24: 6-(Bromomethyl)benzo[b]thiophene-2-carboxylic acid (Int24) [ka]

[0359] To a mixture of methyl 6-(bromomethyl)benzo[b]thiophene-2-carboxylate (Int11b) (5.13 g, 14 mmol) in a mixture of THF and water (4:1, 100 mL), LiOH H 22H2O (623 mg, 15 mmol) was added. The mixture was stirred at room temperature for 4.5 h. EtOAc (100 mL) was added and the organic layer was extracted with 0.1 M NaOH (30 mL). The combined aqueous layers were adjusted to pH = 4 using 1 M HCl and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with anhydrous MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness, and the residue was purified by column chromatography on silica gel to give the title compound.

[0360] Intermediate 24 / 1: 6-(Bromomethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int24 / 1) [ka]

[0361] The title compound was prepared similarly as described for Int24 using ethyl 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylate (Int9f) instead of methyl 6-(bromomethyl)benzo[b]thiophene-2-carboxylate.

[0362] Intermediate 25: 7-Fluoro-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (Int25) [ka]

[0363] 7-Fluoro-2,3-dihydrobenzo[b][1,4]dioxine-6-carbonitrile (30 mg, 0.167 mmol) was dissolved in MeOH (1 mL). To the mixture was added 2M NaOH (1.6 mL, 3.2 mmol). The mixture was stirred at 75° C. overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC.

[0364] Intermediate 26: 7-Fluoro-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (Int26) [ka]

[0365] The title compound was prepared similarly as described for Int2, using 7-fluoro-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid (Int25) in step 1 instead of 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxylic acid.

[0366] In the following compounds of the invention, the examples are named using the example numbers.

[0367] Example 1: N-(3-(benzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 1) [ka]

[0368] To a solution of N-(3-amino-4-methylphenyl)-2,3-dihydro-1,4-benzodioxine-6-carboxamide (Int1) (100 mg, 0.352 mmol), benzo[b]thiophene-2-carboxylic acid (62.7 mg, 0.352 mmol), DIPEA (0.123, 0.703 mmol) and DMAP (6.40 mg, 0.053 mmol) in pyridine (0.980 mL) was added POCl 3 (0.035 mL, 0.387 mmol) was added at 0° C. The mixture was stirred at 0° C. for 15 min. The mixture was then warmed to rt and diluted with EtOAc (15 mL). The organic layer was washed with 0.2 M HCl (3×15 mL) and then saturated NaHCO 3 (3×15 mL). The organic layer was washed with MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d6 )δ ppm:δ 10.19(s,1H),10.06(s,1H),8.33(s,1H),8.07(d,J=7.8Hz,1H),8.01(d,J=7.2Hz,1H),7.87(s,1H),7.61(d,J=8.2H) z,1H),7.57-7.45(m,4H),7.25(d,J=8.4Hz,1H),6.99(d,J=8.3Hz,1H),4.41-4.22(m,4H),2.24(s,3H).MS(ESI):m / z 445.4[M+H] + .

[0369] (Example 1 / 1 to 1 / 18) The following examples were prepared similarly as described for Example 1 using the appropriate building blocks.

[0370] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13] [Table 1-14] [Table 1-15] [Table 1-16] [Table 1-17]

[0371] Example 2: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-methylphenyl)-6-hydroxy-1H-indole-2-carboxamide (Example 2) [ka]

[0372] To a solution of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) (30.0 mg, 0.106 mmol) and 6-hydroxyindole-2-carboxylic acid (18.7 mg, 0.106 mmol) in DMF (0.500 mL) was added NMI (0.025 mL, 0.317 mmol). The mixture was stirred at room temperature for 10 min. PyBroP (54.1 mg, 0.116 mmol) was added and the mixture was heated at 75 °C for 18 h. The mixture was concentrated to dryness and the residue was dissolved in EtOAc (30 mL). The organic layer was washed with 0.1 M HCl (3 x 15 mL) and saturated aqueous NaHCO 3 (3×15 mL). The organic layer was washed with MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was purified by preparative HPLC to give the title compound as a white solid.

[0373] 1 H NMR (300MHz, MeOD-d 3 )δ ppm:δ 7.77(s,1H),7.53(d,J=8.1Hz,1H),7.50-7.43(m,3H),7.27(d,J=8.3Hz,1H),7.21(s,1H),6.94( d,J=8.3Hz,1H),6.86(s,1H),6.70(d,J=8.7Hz,1H),4.36-4.25(m,4H),2.31(s,3H).MS(ESI):m / z 442.2[MH] - .

[0374] (Example 2 / 1) The following examples were prepared similarly as described for Example 2 using the appropriate building blocks. [Table 2]

[0375] Example 3: N-(4-methyl-3-(6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 3) [ka]

[0376] To a mixture of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) (15.0 mg, 0.053 mmol), 6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxylic acid (Int4) (15.1 mg, 0.053 mmol) in DMF (530 μL) was added NMI (13.0 μL, 0.158 mmol). After 5 min, TCFH (16.3 mg, 0.058 mmol) was added. The mixture was stirred at rt for 1 h. The residue was then concentrated to dryness and purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, MeOD-d 3 )δ ppm:δ 8.58(s,1H),8.56(s,1H),8.06(s,1H),7.95-7.80(m,2H),7.74(s,1H),7.65 (d,J=7.9Hz,1H),7.57(s,1H),7.53(d,J=8.4Hz,1H),7.57(s,1H),7.49-7.43 (m,2H),7.39(t,J=6.2Hz,1H),7.27(d,J=8.3Hz,1H),7.19(d,J=8.8,1H),6.9 3(d,J=8.0Hz,1H),5.28(s,2H),4.36-4.24(m,4H),2.30(s,3H).MS(ESI):m / z 552.4[M+H] + .

[0377] (Example 3 / 1 to 3 / 16) The following examples were prepared similarly as described for Example 3 using the appropriate building blocks.

[0378]

Table 3-1

Table 3-2

Table 3-3

Table 3-4

Table 3-5

Table 3-6

Table 3-7

Table 3-8

Table 3-9

Table 3-10

Table 3-11

Table 3-12

Table 3-13

Table 3-14

Table 3-15

Table 3-16

Table 3-17

Table 3-18

Table 3-19

Table 3-20

Table 3-21

Table 3-22

Table 3-23

Table 3-24

Table 3-25

Table 3-26

Table 3-27

Table 3-28

Table 3-29

Table 3-30

Table 3-31

Table 3-32

Table 3-33

Table 3-34

Table 3-35

Table 3-36

[0379] Example 4: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-methylphenyl)thieno[2,3-b]pyrazine-6-carboxamide (Example 4) [ka]

[0380] Thionyl chloride (5.00 mL) was added to thieno[2,3-b]pyrazine-6-carboxylic acid (15.2 mg, 0.059 mmol). The mixture was stirred at 50° C. for 1 h. After the mixture was completed, the mixture was concentrated to dryness. Toluene (2 mL) was added to the concentrated mixture, which was concentrated to dryness. Coevaporation was repeated two more times, and then the residue was dissolved in THF (2 mL). A solution of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) (20.0 mg, 0.070 mmol) and DIPEA (74.0 μL, 0.352 mmol) in THF (2.00 mL) was prepared and brought to 0° C. The thieno[2,3-b]pyrazine-6-carboxylic acid mixture in THF was then added to the cooled Int1 mixture. It was allowed to reach rt and stirred for 18 h. The mixture was then concentrated to dryness and dissolved in EtOAc (20 mL). The resulting mixture was washed with 2M HCl (15.0 mL). The combined organic layers were washed with MgSO 4 The residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, MeOD-d 3 )δ ppm:δ 8.81(s,1H),8.69(s,1H),8.33(s,1H),7.82(s,1H),7.56(d,J=8.4,1H),7.52-7.43(m,2H) ,7.32(d,J=8.4Hz,1H),6.96(d,J=8.2Hz,1H),4.37-4.28(m,4H),2.34(s,3H).MS(ESI):m / z 447.2[M+H] + .

[0381] Example 5: N-(3-(5-(2-hydroxy-2-methylpropoxy)benzofuran-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 5) [ka]

[0382] Step 1: 5-(2-oxopropoxy)benzofuran-2-carboxylic acid (5a) A solution of 5-hydroxybenzofuran-2-carboxylic acid (25.0 mg, 0.140 mmol) in acetone (2.00 mL) was added to the 2 CO 3 (120 mg, 0.870 mmol) was added. After 30 min, 1-chloropropan-2-one (69.0 μL, 0.870 mmol) and NaI (25.2 mg, 0.170 mmol) were added. The mixture was stirred at 65° C. for 72 h. 2 CO 3 (41.9 mg, 0.300 mmol) and water (0.500 mL) were added to the mixture. The mixture was stirred at 45 °C for 4.5 h. Then the mixture was cooled to rt and 1M HCl (10 mL) was added. The resulting mixture was extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with MgSO 4 Dry on ice, filter and concentrate to dryness to give the crude title compound.

[0383] Step 2: N-(4-methyl-3-(5-(2-oxopropoxy)benzofuran-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (5b) Thionyl chloride (2 mL) was added to 5-(2-oxopropoxy)benzofuran-2-carboxylic acid (5a) (28.0 mg, 0.120 mmol). The mixture was stirred at 53° C. for 1 h. After the mixture was completed, the mixture was concentrated to dryness. Toluene (2 mL) was added to the concentrated mixture, which was concentrated to dryness. Coevaporation was repeated two more times, and then the crude product was dissolved in THF (2 mL). A solution of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) (34.0 mg, 0.120 mmol) and DIPEA (42.0 μL, 0.240 mmol) in THF (2.00 mL) was prepared and brought to 0° C. The crude 5a mixture in THF was then added to the cooled Int1 mixture. The mixture was brought to rt and stirred for 18 h. The mixture was then concentrated to dryness in vacuo and dissolved in EtOAc (20 mL). The resulting mixture was washed with 2M HCl (15 mL). The combined organic layers were washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound.

[0384] Step 3: N-(3-(5-(2-hydroxy-2-methylpropoxy)benzofuran-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 5) To a solution of N-(4-methyl-3-(5-(2-oxopropoxy)benzofuran-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (5b) (65.0 mg, 0.112 mmol) in THF (1.00 mL) was added methylmagnesium bromide (1.0 M in THF, 0.503 mL, 0.503 mmol) at 0 °C. Then water (2 mL) was added. The resulting mixture was extracted with EtOAc (3 × 4 mL). The combined organic layers were washed with MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300 MHz, CDCl 3)δ ppm:δ 8.26(s,1H),8.17(s,1H),7.84(s,1H),7.77(d,J=8.3Hz,1H), 7.52(s,1H),7.48(d,J=8.7Hz,1H),7.44(s,1H),7.37(d,J=8.5Hz,1H),7.22(s,1H),7.13-7.10(m ,2H),6.93(d,J=8.4,1H),4.34-4.26(m,4H),3.85(s,2H),2.39(s,3H),1.38(s,6H).MS(ESI):m / z 517.5[M+H] + .

[0385] Example 6: N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) [ka]

[0386] BBr 3 (1.0M solution in DCM, 7.9 mL, 0.800 mmol) was added dropwise to a solution of Example 1 / 12 (0.750 mg, 0.200 mmol) in DCM (15.8 mL) at -78°C. The mixture was stirred at -78°C for 1 h and then slowly warmed to rt for 18 h. EtOH (1.00 mL) was added dropwise at 0°C and the mixture was allowed to reach rt. This was diluted with 1M HCl (3 x 30 mL), saturated aqueous NaHCO 3 (3×30 mL), and then brine (30 mL). The organic layer was washed with MgSO 4 The mixture was then purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6)δ ppm:δ 10.04(s,1H),10.0(s,1H),9.94(s,1H),8.17(s,1H),7.84(s,1H),7.79(d,J=8.7Hz,1H),7.59(d,J=8.3Hz,1H),7.5 5-7.47(m,2H),7.32(s,1H),7.23(d,J=8.3Hz,1H),7.02-6.93(m,2H),4.37-4.27(m,4H),2.23(s,3H).MS(ESI):m / z 461.4[M+H] + .

[0387] Example 7: N-(3-(6-(2-methoxyethoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 7) [ka]

[0388] N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) (17.5 mg, 0.038 mmol) and K 2 CO 3 To a solution of (10.5 mg, 0.076 mmol) in DMF (0.380 mL) was added 2-bromoethyl methyl ether (5.80 mg, 0.042 mmol). The mixture was stirred at 120° C. for 1 h. The mixture was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6)δ ppm:δ 10.06(s,1H),10.05(s,1H),8.23(s,1H),7.92-7.82(m,2H),7.63(s,1 H),7.59(d,J=8.6Hz,1H),7.55-7.48(m,2H),7.24(d,J=8.3Hz,1H),7. 10(d,J=8.8,1H),6.98(d,J=8.3Hz,1H),4.37-4.27(m,4H),4.25-4.16(m,2H),3.71(t,J=4.4Hz,2H),3.34(s,3H),2.23(s,3H).MS(ESI):m / z 519.5[M+H] + .

[0389] Example 8: N-(4-methyl-3-(6-(2-oxopropoxy)benzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 8) [ka]

[0390] N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) (15.0 mg, 0.033 mmol) and K 2 CO 3 To a solution of (3.00 μL, 0.039 mmol) in DMF (0.326 mL) was added chloroacetone (3.00 μL, 0.039 mmol). The mixture was stirred at 100° C. for 1 h. The mixture was then purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6)δ ppm:δ 10.08(s,1H),10.04(s,1H),8.23(s,1H),7.93-7.82(m,2H),7.66-7.48(m,4H),7.24(d,J=8.4Hz,1H),7.11(d ,J=8.8Hz,1H),6.98(d,J=8.2Hz,1H),4.94(s,2H),4.38-4.26(m,4H),2.23(s,3H),2.20(s,3H).MS(ESI):m / z 515.2[MH] + .

[0391] Example 9: N-(3-(6-(2-hydroxy-2-methylpropoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 9) [ka]

[0392] To a solution of N-(4-methyl-3-(6-(2-oxopropoxy)benzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 8) (10.0 mg, 0.019 mmol) in THF (0.190 mL) was added MeMgBr (2 M, 0.015 mL, 0.031 mmol) at 0° C. and then warmed to rt. After 1 h, saturated aqueous NH 4 Cl (1 mL) was added to the mixture, which was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300 MHz, CDCl 3)δ ppm:δ 8.03(s,1H),7.86(s,1H),7.80(s,1H),7.77-7.62(m,3H),7.42(s,1H),7.40-7.29(m,2H),7.19(d,J=8.3Hz,1H),7 .08(d,J=8.9Hz,1H),6.92(d,J=8.3Hz,1H),4.35-4.24(m,4H),3.89(s,2H),2.32(s,3H),1.39(s,6H).MS(ESI):m / z 533.4[M+H] + .

[0393] Example 10: N-(3-(6-(2-hydroxyethoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 10) [ka]

[0394] Step 1: Ethyl 2-bromoacetate (10a) To a solution of 2-bromoethanol in DCM (16.0 mL) was added acetyl chloride (0.683 mL, 9.67 mmol) and then TEA (0.728 mL, 10.6 mmol). The mixture was stirred at room temperature for 72 h. The mixture was diluted with DCM (50 mL) and saturated aqueous NH 4 Cl (20 mL) and then saturated aqueous NaHCO 3 (20 mL). The organic layer was washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the crude title compound as a yellow liquid.

[0395] Step 2: Methyl 6-(2-acetoxyethoxy)benzo[b]thiophene-2-carboxylate (10b) Ethyl 6-methoxybenzo[b]thiophene-2-carboxylate (Int4b) (200 mg, 0.960 mmol) and Cs 2 CO 3To a solution of (250 mg, 1.15 mmol) in DMF (3.20 mL) was added crude ethyl 2-bromoacetate (10a) (250 mg, 1.15 mmol). The mixture was stirred at 60° C. for 1 h. The solvent was removed under reduced pressure to give the crude title compound.

[0396] Step 3: 6-(2-hydroxyethoxy)benzo[b]thiophene-2-carboxylic acid (10c) To a solution of methyl 6-(2-acetoxyethoxy)benzo[b]thiophene-2-carboxylate (10b) (283 mg, 0.960 mmol) in MeOH (4.00 mL) was added 2M aqueous NaOH (1.00 mL). It was brought to reflux and stirred for 1 h. The mixture was cooled to rt and the pH was adjusted to about 2 with 1M HCl. The mixture was then extracted with EtOAc (3×30 mL). The organic layers were combined, washed with brine and diluted with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound as a light brown solid.

[0397] Step 4: N-(3-(6-(2-hydroxyethoxy)benzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 10) To a mixture of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) (298 mg, 1.05 mmol), 6-(2-hydroxyethoxy)benzo[b]thiophene-2-carboxylic acid (10c) (100 mg, 0.420 mmol) in DMF (4.20 mL) was added NMI (0.117 mL, 1.47 mmol). After 5 min, TCFH (130 mg, 1.47 mmol) was added. The mixture was stirred for 15 min at rt. The residue was then diluted with EtOAc (200 mL) and washed with 1 M HCl (3×30 mL) and saturated aqueous NaHCO 3(30 mL). The organic layer was concentrated to dryness and purified by column chromatography on silica gel (acetone / cyclohexane=1:10). The resulting solid was triturated with MeCN (2×10 mL) and pentane (7×5 mL). It was then suspended in MeCN / water (1:1), frozen, and placed on a lyophilizer to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6 )δ ppm:10.06(s,1H),10.04(s,1H),8.22(s,1H),7.90-7.81(m,2H),7.64-7.56(m,2H),7.55-7.49(m,2H),7.23(d,J=8.2Hz,1H),7.09(d,J=8.2 Hz,1H),6.98(d,J=8.3Hz,1H),4.89(t,J=5.6Hz,1H),4.35-4.27(m,4H),4.10(t,J=4.8Hz,2H),3.80-3.72(m,2H),2.23(s,3H).MS(ESI):m / z 503.2[MH] - .

[0398] Example 11: N-(4-methyl-3-(6-(2-(2-oxopropoxy)ethoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 11) [ka]

[0399] Step 1: 2-(allyloxy)ethyl 4-methylbenzenesulfonate (11a) To a mixture of 2-allyloxyethanol (200 mg, 1.96 mmol) in DCM (20.0 mL) was added sodium hydroxide (411 mg, 2.15 mmol) and 4-toluenesulfonyl chloride (313 mg, 7.84 mmol) at 0° C., then the solution was stirred for 0.5 h. The solution was stirred at 25° C. for 4 h. The mixture was washed with water (3×20 mL). The organic layer was extracted with Na 2 SO 4It was dried over ice, filtered and concentrated to dryness to give the crude title compound.

[0400] Step 2: N-(3-(6-(2-(allyloxy)ethoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (11b) N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) (45.0 mg, 0.097 mmol) and K 2 CO 3 To a solution of (27.0 mg, 0.195 mmol) in DMF (0.977 mL) was added 2-(allyloxy)ethyl 4-methylbenzenesulfonate (11a) (27.6 mg, 0.107 mmol). Stirred at 120° C. for 18 h. The solvent was removed under reduced pressure to give the crude title compound.

[0401] Step 3: N-(4-methyl-3-(6-(2-(2-oxopropoxy)ethoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 11) To a stirred solution of N-(3-(6-(2-(allyloxy)ethoxy)benzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (11b) (30.0 mg, 0.055 mmol) in MeCN (1.00 mL) and water (0.150 mL) was added Pd(OAc) 2 (0.600 mg, 0.003 mmol) and DMP (28.0 mg, 0.066 mmol) were added at room temperature. The mixture was warmed to 50° C. and stirred for 18 h. It was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300 MHz, CDCl 3)δ ppm:δ 8.09(s,1H),7.84-7.69(m,4H),7.64(s,1H),7.43(s,1H),7.40-7.31(m,2H),7.22(s,1H),7.09(d,J=8.9Hz,1H),6 .94(d,J=8.5Hz,1H),4.37-4.24(m,6H),4.21(s,2H),3.95(t,J=4.8Hz,2H),2.35(s,3H),2.19(s,3H).MS(ESI):m / z 561.5[M+H] + .

[0402] Example 12: N-(4-methyl-3-(6-(2-(pyrrolidin-1-yl)ethoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 12) [ka]

[0403] To a solution of N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) (20.0 mg, 0.043 mmol) and NaH (1.90 mg, 0.086 mmol) in DMF (0.391 mL) was added 1-(2-chloroethyl)pyrrolidine hydrochloride (7.3 mg, 0.047 mmol). The mixture was stirred at 50° C. for 18 h. It was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6)δ ppm:δ 10.06(s,1H),10.05(s,1H),8.22(s,1H),7.92-7.81(m,2H),7.63(s,1H) ,7.59(d,J=8.0,1H),7.56-7.47(m,2H),7.24(d,J=8.4Hz,1H),7.09(d,J =8.8Hz,1H),6.98(d,J=8.3Hz,1H),4.36-4.27(m,4H),4.18(t,J=5.8Hz,2H),2.84(t,J=6.0Hz,2H),2.23(s,3H),1.77-1.65(m,4H).MS(ESI):m / z 558.6[M+H] + .

[0404] Example 13: (S)—N-(3-(6-(3-(3-fluoropyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 13) [ka]

[0405] To a solution of N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) (50.0 mg, 0.108 mmol) in DMF (1.10 mL) was added K 2 CO 3 (33.0 mg, 0.238 mmol) and 1-bromo-3-chloropropane (0.013 mL, 0.130 mmol) were added. This was brought to 65° C. and stirred for 18 hours. Then, (S)-3-fluoropyrrolidine hydrochloride (16.4 mg, 0.130 mmol) and K 2 CO 3 (11.5 mg, 0.116 mmol) was added and stirred for 24 h at 65° C. This was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6)δ ppm:δ 10.06(s,1H),10.05(s,1H),8.22(s,1H),7.92-7.81(m,2H),7.65-7.47(m,4H),7.24(d,J=8.4 Hz,1H),7.08(d,J=9.1Hz,1H),6.98(d,J=8.5Hz,1H),5.20(dt,J=13.5,5.4Hz,1H),4.38-4.25( m,4H),4.13(t,J=6.3Hz,2H),2.92-2.74(m,2H),2.73-2.74(m,1H),2.63-2.54(m,2H),2.39-2 .28(m,1H),2.23(s,3H),2.20-2.01(m,1H),1.99-1.89(m,2H),1.88-1.74(m,1H).MS(ESI):m / z 590.6[M+H] + .

[0406] (Examples 13 / 1 to 13 / 3) The following examples were prepared similarly as described for Example 13 using the appropriate building blocks.

[0407] [Table 4-1] [Table 4-2] [Table 4-3]

[0408] Example 14: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-methylphenyl)-1-methyl-1H-indole-2-carboxamide (Example 14) [ka]

[0409] Step 1: 1-Methyl-1H-indole-2-carboxylic acid (14a) To a mixture of 1H-indole-2-carboxylic acid (10.0 mg, 0.062 mmol) in anhydrous DMF (0.621 mL) was added NaH (4.80 mg, 0.200 mmol) at 0 °C. The mixture was stirred at 0 °C for 15 min, then MeI (12.0 μL, 0.200 mol) was added. The mixture was brought to rt and stirred for 18 h. Then the solvent was removed in vacuum. Water (10 mL) was added to the residue and the pH was adjusted to pH = 3. The resulting mixture was extracted with EtOAc (10 mL x 3). The combined organic layers were washed with MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness to give crude 1-methyl-1H-indole-2-carboxylic acid.

[0410] Step 2: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-methylphenyl)-1-methyl-1H-indole-2-carboxamide (Example 14) To a mixture of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int1) (10.0 mg, 0.035 mmol), 1-methyl-1H-indole-2-carboxylic acid (14a) (7.40 mg, 0.042 mmol) in DMF (0.350 mL) was added NMI (8.00 μL, 0.106 mmol). After 5 min, TCFH (10.9 mg, 0.039 mmol) was added. The mixture was stirred at room temperature for 1 h. The residue was then concentrated to dryness and purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, MeOD-d 3 )δ ppm:δ 7.80(s,1H),7.67(d,J=8.1Hz,1H),7.56-7.43(m,4H),7.37-7.24(m,3H),7.14(t,J=7.5 Hz,1H),6.94(d,J=8.1Hz,1H),4.36-4.24(m,4H),4.05(s,3H),2.33(s,3H).MS(ESI):m / z 442.4[M+H] + .

[0411] Example 15: N-(3-(benzo[b]thiophene-2-carboxamido)-4-methylphenyl)chroman-7-carboxamide (Example 15) [ka]

[0412] Step 1: N-(3-amino-4-methylphenyl)chroman-7-carboxamide (15a) To a mixture of chroman-7-carboxylic acid (50.0 mg, 0.409 mmol), 2,4-diaminotoluene (72.9 mg, 0.409 mmol) in DMF (0.949 mL) was added NMI (98.0 μL, 1.23 mmol). After 5 min, TCFH (149 mg, 0.532 mmol) was added. The mixture was stirred at rt for 1 h. The reaction was diluted with 1 M HCl (40 mL) and washed with EtOAc (3×30 mL). The aqueous layer was adjusted to pH=3, extracted with EtOAc (3×50 mL), and washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound.

[0413] Step 2: N-(3-(benzo[b]thiophene-2-carboxamido)-4-methylphenyl)chroman-7-carboxamide (Example 15) To a mixture of N-(3-amino-4-methylphenyl)chroman-7-carboxamide (15a) (104 mg, 0.368 mmol), 1-benzothiophene-2-carboxylic acid (65.6 mg, 0.368 mmol) in DMF (1.50 mL) was added NMI (88.0 μL, 1.105 mmol). After 5 min, TCFH (134 mg, 0.479 mmol) was added. The mixture was stirred at rt for 1 h. MeCN / water (5 mL) was then added to the mixture and the precipitated solid was triturated with MeCN / water (1:1) to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6)δ ppm:δ 10.20(s,1H),10.02(s,1H),8.33(s,1H),8.07(d,J=7.6,1H),9.01(d,J= 7.4Hz,1H)7.86(s,1H),7.78-7.67(m,2H),7.61(d,J=8.4Hz,1H),7.54-7. 42(m,2H),7.25(d,J=8.4Hz,1H),6.84(d,J=8.4Hz,1H),4.21(t,J=5.1Hz ,2H),2.83(t,J=6.4Hz,2H),2.24(s,3H),2.01-1.91(m,2H).MS(ESI):m / z 443.4[M+H] + .

[0414] Example 16: N-(3-(6-(((1S,4S)-5-ethyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 16) [ka]

[0415] Step 1: N-(3-(6-(bromomethyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (16a) The title compound was prepared similarly as described for Example 3, using 6-(bromomethyl)benzo[b]thiophene-2-carboxylic acid (Int24) instead of 6-(pyridin-2-ylmethoxy)benzo[b]thiophene-2-carboxylic acid, and N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) instead of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide.

[0416] Step 2: (1S,4S)-tert-butyl 5-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)methyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (16b) A solution of N-(3-(6-(bromomethyl)benzo[b]thiophene-2-carboxamido)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (16a) (17.0 mg, 0.031 mmol) in dry DMF (0.300 mL) was treated with (1S,4S)-tert-butyl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (6.2 mg, 0.031 mmol) and Cs 2 CO 3 (11.3 mg, 0.035 mmol) was added. The mixture was stirred at 60 °C for 3 h. The mixture was diluted with EtOAc (20 mL) and washed with 1 M HCl (10 mL) and water (10 mL). The organic layer was washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0417] Step 3: N-(3-(6-(((1S,4S)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (16c) To a solution of (1S,4S)-tert-butyl 5-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)methyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate (16b) (20 mg, 0.030 mmol) in dry DCM (0.300 mL) was added TFA (0.100 mL) dropwise at 0° C. The mixture was allowed to warm to rt and stirred for 2 h. Saturated NaHCO 3 was added and the aqueous layer was extracted with DCM (3×15 mL). The combined organic layers were diluted with MgSO4 Drying on ice, filtration and concentration to dryness gave the title compound which was used in the next step without further purification.

[0418] Step 4: N-(3-(6-(((1S,4S)-5-ethyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 16) To a solution of N-(3-(6-((1S,4S)-2,5-diazabicyclo[2.2.1]heptan-2-ylmethyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (16c) (8.0 mg, 0.014 mmol) in dry DMF (0.143 mL) was added NaBH 3 CN (0.9 mg, 0.014 mmol) was added and the mixture was cooled to 0° C. Acetaldehyde (0.001 mL, 0.014 mmol) was added as a solution in 0.5 mL DMF. The mixture was stirred for 5 h. Saturated NaHCO 3 (10 mL) was added and the aqueous layer was extracted with EtOAc (3×15 mL). The combined organic layers were diluted with MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, CD 3 OD):δ ppm 8.19-8.12(m,2H),7.98(s,1H),7.94(d,J=9Hz,1H),7.62-7.54(m,1H) ,7.54-7.46(m,3H),7.23(t,J=9.6Hz,1H),6.96(d,J=8.2Hz,1H),4.36- 4.29(m,4H),4.03-3.80(m,3H),3.55(s,2H),3.12-2.76(m,5H),2.82(d ,J=11.5Hz,1H),2.11-1.89(m,2H),1.24(t,J=7.1Hz,3H).MS(ESI):m / z 587.6[M+H] + .

[0419] Example 17: N-(3-(6-((4-ethyl-1,4-diazepan-1-yl)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 17) [ka]

[0420] Step 1: tert-Butyl 4-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)methyl)-1,4-diazepane-1-carboxylate (17a) The title compound was prepared similarly as described for Example 3, using 6-((4-(tert-butoxycarbonyl)-1,4-diazepan-1-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int11 / 4) instead of 6-((pyridin-2-yloxy)methyl)benzo[b]thiophene-2-carboxylic acid, and N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) instead of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide.

[0421] Step 2: N-(3-(6-((1,4-diazepan-1-yl)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (17b) To a solution of tert-butyl 4-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)methyl)-1,4-diazepane-1-carboxylate (17a) (45.6 mg, 0.069 mmol) in DCM (0.690 mL) was added TFA (0.200 mL) dropwise at 0 °C. The mixture was allowed to warm to rt. After 2 h, the reaction was quenched with saturated NaHCO 3 The mixture was quenched with 10 mL of DCM. The aqueous layer was extracted with DCM (3×20 mL). The combined organic layers were washed with anhydrous MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound.

[0422] Step 3: N-(3-(6-((4-ethyl-1,4-diazepan-1-yl)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 17) To a solution of N-(3-(6-((1,4-diazepan-1-yl)methyl)benzo[b]thiophene-2-carboxamido)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (17b) (24.0 mg, 0.043 mmol) in dry DMF (0.214 mL) was added Cs 2 CO 3 (13.9 mg, 0.043 mmol) followed by iodoethane (0.003 mL, 0.043 mmol) were added at room temperature. The mixture was stirred for 18 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, CD 3OD):δ ppm 11.22(s,1H),10.98(s,1H),9.15(s,1H),8.92(d,J=9Hz,1H),8.80-8. 72(m,2H),8.50-8.42(m,1H),8.38-8.22(m,3H),8.10(t,J=9.3Hz,1H) ,7.80(d,J=8.1Hz,1H),5.13(m,4H),4.56(s,2H),4.05-3.90(m,4H),3.55-3.40(m,6H)2.60-2.50(m,2H),1.80(t,J=6.9Hz,3H)MS(ESI):m / z 589.6[M+H] + .

[0423] Example 18: (R)-N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-((3-hydroxypyrrolidin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 18) [ka]

[0424] Step 1: 6-(bromomethyl)-N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)thieno[2,3-b]pyridine-2-carboxamide (18a) The title compound was prepared similarly as described for Example 3, using 6-(bromomethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int24 / 1) instead of 6-((pyridin-2-yloxy)methyl)benzo[b]thiophene-2-carboxylic acid, and N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) instead of N-(3-amino-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide.

[0425] Step 2: (R)-N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)-6-((3-hydroxypyrrolidin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 18) To a solution of 6-(bromomethyl)-N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)thieno[2,3-b]pyridine-2-carboxamide (18a) (10.0 mg, 0.018 mmol) in dry DMF (0.184 mL) was added Et 3 N (0.003 mL, 0.020 mmol) and (R)-pyrrolidin-3-ol (1.7 mg, 0.019 mmol) were added. The mixture was stirred at 60° C. for 18 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, CD 3 OD):δ ppm 8.35(d,J=8.2Hz,1H),8.18-8.10(m,2H),7.61-7.51(m,2H),7.49-7.41(m,2H),7.21(t,J=9.5Hz,1H),6.93(d,J=8.2Hz,1H),4.48 (s,1H),4.35(s,2H),4.32-4.25(m,4H),3.32-3.17(m,2H),3.16-2.96(m,2H),2.32-2.11(m,1H),1.99-1.86(m,1H).MS(ESI):m / z 549.5[M+H] + .

[0426] Example 18 / 1: (S)-N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-((3-hydroxypyrrolidin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 18 / 1) [ka]

[0427] The title compound was prepared similarly as described for Example 18, using (S)-pyrrolidin-3-ol in place of (R)-pyrrolidin-3-ol in step 2. 1 H NMR (300MHz, CD 3 OD):δ ppm:8.38(d,J=8.2Hz,1H),8.20-8.10(m,2H),7.62-7.52(m,2H),7.50-7.43(m,2H),7.22(t,J=9.5Hz,1H),6.94(d,J=8.2Hz,1H),4. 52(s,1H),4.45(s,2H),4.34-4.27(m,4H),3.48-3.34(m,2H),3.30-3.08(m,2H),2.34-2.18(s,1H),2.02-1.93(m,1H).MS(ESI):m / z 549.5[M+H] + .

[0428] Example 18 / 2: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-((4-hydroxypiperidin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 18 / 2) [ka]

[0429] The title compound was prepared similarly as described for Example 18, using piperidin-4-ol in step 2 instead of (3R)-3-pyrrolidinol. 1 H NMR (300MHz, CD 3OD):δ ppm 8.37(d,J=7.5Hz,1H),8.19-8.10(m,2H),7.63-7.51(m,2H),7.50-7.43(m,2H),7.21(t,J=9.6Hz,1H),6.94(d,J=8.3Hz,1H),4.34-4 .25(m,4H),4.21(s,2H),3.88-3.77(m,1H),3.26-3.15(m,2H),2.88-2.76(m,2H),2.06-1.93(m,2H),1.83-1.67(m,2H).MS(ESI):m / z 563.5[M+H] + .

[0430] Example 19: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxamide (Example 19) [ka]

[0431] To a solution of 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15) (260 mg, 0.89 mmol) and N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (257 mg, 0.89 mmol) in acetonitrile (10 mL) was added NMI (255 mg, 3.11 mmol) and TCFH (448 mg, 1.6 mmol) and the mixture was stirred at 25° C. for 3 h. The mixture was poured into water. The mixture was extracted with EtOAc (3×10 ml). The combined organic layers were washed with brine and diluted with anhydrous Na 2 SO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was purified by reverse phase column chromatography (solvent A:H 2 0, solvent B: MeCN) to give the title compound. 1 H NMR (400MHz, CD 3OD):δ ppm 8.23(d,J=8.8Hz,1H),8.14(dd,J=6.8,2.4Hz,1H),8.08(s,1H),7.56-7.52(m,1H),7.49-7.45(m,2H),7.25-7.17(m,1H),7.00(d,J=8.8H) z,1H),6.94(d,J=8.0Hz,1H),4.78-4.71(m,2H),4.35-4.27(m,4H),3.63-3.56(m,2H),3.38-3.34(m,4H),2.10-2.06(m,4H).MS(ESI):m / z 563.7[M+H] + .

[0432] Example 19 / 1: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-(3-(pyrrolidin-1-yl)propoxy)thieno[2,3-b]pyridine-2-carboxamide (Example 19 / 1) [ka]

[0433] The title compound was prepared similarly as described for Example 19, using 6-(3-(pyrrolidin-1-yl)propoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15 / 1) instead of 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15). 1 H NMR (400MHz, DMSO-d 6 ):δ ppm 10.45(s,1H),10.18(s,1H),8.35-8.29(m,2H),8.13-8.11(m,1H),7.63-7. 50(m,3H),7.32-7.28(m,1H),7.01-6.97(m,2H),4.46(t,J=4.0Hz,2H),4.33 -4.30(m,4H),3.70-3.40(m,2H),3.33-3.30(m,2H),3.20-2.98(m,2H),2.20-2.15(m,2H),2.08-2.95(m,4H).MS(ESI):m / z 577.6[M+H] +.

[0434] Example 19 / 2: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-(2-(4-ethylpiperazin-1-yl)ethyl)thieno[2,3-b]pyridine-2-carboxamide (Example 19 / 2) [ka]

[0435] The title compound was prepared similarly as described for Example 19, using 6-(2-(4-ethylpiperazin-1-yl)ethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int19) instead of 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15). 1 H NMR (400MHz, DMSO-d 6 ):δ ppm 10.52(s,1H),10.18(s,1H),8.34-8.31(m,2H),8.11-8.09(m,1H),7.54 -7.53(m,1H),7.47-7.45(m,3H),7.30(t,J=8Hz,1H),7.01(d,J=8Hz,1H),4.32-4.31(m,4H) ,3.06-3.02(m,2H),2.73-2.70(m,2H),2.34-2.26(m,10H),0.98(t,J=8Hz,3H).MS(ESI):m / z 590.1[M+H] + .

[0436] Example 19 / 3: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-((1-ethylpiperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 19 / 3) [ka]

[0437] The title compound was prepared similarly as described for Example 19, using 6-((1-ethylpiperidin-4-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int20) instead of 6-(2-(pyrrolidin-1-yl)ethoxy)thieno[2,3-b]pyridine-2-carboxylic acid (Int15). 1 H NMR (400MHz, DMSO-d 6 ):δ ppm 10.52(s,1H),10.19(s,1H),8.33-8.31(m,2H),8.12-8.11(m,1H),7.69- 7.67(m,1H),7.66-7.53(m,2H),7.51-7.31(m,2H),7.28-6.99(m,1H),4. 33-4.30(m,4H),2.83-2.79(m,4H),2.27-2.24(m,2H),1.82-1.76(m,3H) ,1.57-1.54(m,2H),1.29-1.24(m,2H),0.96(t,J=8Hz,3H).MS(ESI):m / z 575.2[M+H] + .

[0438] Example 20: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxamide (Example 20) [ka]

[0439] To a solution of 6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxylic acid (Int21) (0.7 g, 2.4 mmol) and N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (490 mg, 1.7 mmol) in DMF (10 mL) was added triethylamine (0.3 ml, 2.1 mmol) and HATU (323 mg, 0.85 mmol). The mixture was stirred at 50° C. for 6 h. The mixture was concentrated to dryness and the residue was purified by reverse phase column chromatography (H 2 O / MeOH=40:60) to give the crude product, which was further purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (400MHz, CD 3 OD):δ ppm 8.57(s,1H),8.11(dd,J=6.9,2.6Hz,1H),7.72(s,1H),7.60-7.45(m,3H),7.24-7.17(m,1H),6.94(d, J=8.3Hz,1H),4.36-4.24(m,4H),3.64(s,2H),2.80-2.43(m,10H),1.10(t,J=7.2Hz,3H).MS(ESI):m / z 565.2[M+H] + .

[0440] Example 20 / 1: N-(4-fluoro-3-(6-((pyridin-2-yloxy)methyl)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 20 / 1) [ka]

[0441] The title compound was prepared similarly as described for Example 20, using 6-((pyridin-2-yloxy)methyl)benzo[b]thiophene-2-carboxylic acid (Int16) instead of 6-((4-ethylpiperazin-1-yl)methyl)imidazo[2,1-b]thiazole-2-carboxylic acid (Int21). 1 H NMR (400MHz, DMSO-d 6 ):δ ppm 10.47(s,1H),10.17(s,1H),8.36(s,1H),8.20(dd,J=5.0,1.3Hz,1H),8 .14(s,1H),8.11-8.08(m,1H),8.02(d,J=8.3Hz,1H),7.78-7.72(m,1H), 7.69-7.63(m,1H),7.57-7.50(m,3H),7.35-7.27(m,1H),7.04-6.98(m, 2H),6.93(d,J=8.3Hz,1H),5.50(s,2H),4.33-4.30(m,4H).MS(ESI):m / z 556.2[M+H] + .

[0442] Example 21: N-(3-(6-(1-(4-ethylpiperazin-1-yl)ethyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 21) [ka]

[0443] Step 1: 6-(1-hydroxyethyl)benzo[b]thiophene-2-carboxylic acid (21a) To a solution of 6-bromobenzo[b]thiophene-2-carboxylic acid (50.0 mg, 0.194 mmol) in dry THF was added 2.5 M n-BuLi in hexane (0.163 mL, 0.408 mmol) at -78 °C. The mixture was stirred at that temperature for 30 min. Acetaldehyde (0.009 mL, 0.214 mmol) was added dropwise as a solution in 0.214 mL of dry THF. The mixture was allowed to warm slowly to 0 °C in an ice bath and stirred for 1 h. Saturated aqueous NH 4 Cl (1 mL) was added to the mixture. The mixture was then cooled to H 2 The mixture was diluted with 2×O (10 mL) and the pH was adjusted to approximately 2 with 1 M HCl. The aqueous layer was extracted with EtOAc (2×20 mL). The combined organic layers were washed with brine and MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was purified by preparative HPLC to give the title compound as a white solid.

[0444] Step 2: 6-(1-chloroethyl)benzo[b]thiophene-2-carboxylic acid (21b) To a solution of 6-(1-hydroxyethyl)benzo[b]thiophene-2-carboxylic acid (21a) (4.5 mg, 0.020 mmol) in dry DCM (0.20 mL) was added SOCl 2 (3 μL, 0.040 mmol) was added and the mixture was stirred for 4 h. The mixture was concentrated to dryness and the residue was used in the next step without further purification.

[0445] Step 3: N-(3-(6-(1-(4-ethylpiperazin-1-yl)ethyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 21) To a solution of 6-(1-chloroethyl)benzo[b]thiophene-2-carboxylic acid (21b) (4.8 mg, 0.020 mmol) and N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (5.8 mg, 0.020 mmol) in dry DMF (0.1 mL) was added TCFH (5.6 mg, 0.020 mmol) and NMI (0.006 mL, 0.070 mmol). The mixture was stirred at rt for 30 min. To the mixture was added 1-ethylpiperazine (0.005 mL, 0.040 mmol) and the mixture was stirred at 60° C. for 18 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300MHz, CD 3 OD):δ ppm:8.18-8.07(m,2H),7.95-7.89(m,2H),7.60-7.53(m,1H),7.51-7.42(m,3H),7.21(t,J=9.5Hz,1H),6.94(d,J=8.2Hz,1H),4.34-4. 26(m,4H),3.61(q,J=6.6Hz,1H),2.84-2.47(m,10H),2.17(d,J=7.3Hz,1H),1.47(d,J=6.5Hz,3H),1.14(t,J=7.2Hz,3H).MS(ESI):m / z 589.6[M+H] + .

[0446] Example 22: N-(3-(6-(2-aminoethoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 22) [ka]

[0447] N-(3-(6-hydroxybenzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 6) (10.0 mg, 0.022 mmol) and K 2CO 3 To a solution of (8.5 mg, 0.026 mmol) in DMF (0.217 mL) was added 2-(Boc-amino)ethyl bromide (5.4 mg, 0.024 mmol). The mixture was stirred at 60° C. for 18 h. The mixture was concentrated to dryness in vacuo and the residue was redissolved in EtOAc (20 mL). The resulting mixture was washed with saturated NH 4 The combined organic layers were washed with MgSO 4 The mixture was dried over ice, filtered and concentrated to dryness. The residue was dissolved in dry DCM (0.083 mL) and TFA (0.038 mL, 0.497 mmol) was added dropwise at 0° C. The mixture was brought to rt and stirred for 3 h. The mixture was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (300 MHz, CDCl 3 ):δ ppm 8.08(s,1H),7.84-7.60(m,5H),7.43(s,1H),7.40-7.30(m,2H),7.07(d,J=9.2Hz,1),6.94(d,J=8.6Hz,1H),4.37 -4.24(m,4H),4.13-4.02(m,2H),3.15(m,2H),2.35(s,3H).MS(ESI):m / z 504.5[M+H] + .

[0448] Example 23: 6-((4-acetylpiperazin-1-yl)methyl)-N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)thieno[2,3-b]pyridine-2-carboxamide (Example 23) [ka]

[0449] To a mixture of 6-(piperazin-1-ylmethyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int10 / 4) (30.0 mg, 0.108 mmol) in dry DMF (2 mL) was added TEA (15 μL, 0.108 mmol) and acetyl chloride (8 μL, 0.108 mmol) and the mixture was stirred at room temperature for 2 h. N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (31.2 mg, 0.108 mmol) and NMI (26 μL, 0.325 mmol) were added and the mixture was stirred at room temperature for 5 min. TCFH (30.4 mg, 0.108 mmol) was added in portions and the mixture was stirred at rt overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.55(s,1H),10.17(s,1H),8.48-8.31(m,2H),8.15-8.10(m,1H),7.68-7.61(m,2H),7.55-7.50(m,2H),7.30(t,J=9.7Hz,1H),6.99 (d,J=8.4Hz,1H),4.35-4.27(m,4H),3.86-3.70(m,2H),3.46(s,2H),3.25-3.02(m,2H),2.47-2.28(m,4H),2.00(s,3H).MS(ESI):m / z 590.5[M+H] + .

[0450] Example 24: N-(3-(6-(2-(4-ethylpiperazin-1-yl)ethoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 24) [ka]

[0451] To a mixture of N-(3-(6-(2-hydroxyethoxy)benzo[b]thiophene-2-carboxamido)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30 mg, 0.059 mmol) (Example 10) in dry DMF (2 mL) was added sodium hydride (2.9 mg, 0.071 mmol) (60% suspension in mineral oil) and the mixture was stirred at room temperature for 30 min. Then p-TsCl (12.5 mg, 0.065 mmol) was added and the mixture was stirred at room temperature for 30 min. 1-Ethylpiperazine (8 μL, 0.060 mmol) was added and the mixture was stirred at room temperature overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC (with a gradient of 40-65% MeCN in water) to give the title compound as a white solid. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.06(s,1H),10.03(s,1H),8.22(s,1H),7.86(s,1H),7.63(s,1H),7.55 -7.51(m,1H),7.48-7.45(m,2H),7.23(d,J=8.1Hz,1H),7.11-7.08(m,2H),6.97(d,J=8.5Hz,1H),4.34-4.27(m,4H),4. 23-4.17(m,2H),3.18-2.76(m,8H),2.74-2.70(m,2H),2.29-2.27(m,2H),2.21(s,3H),1.21-1.10(m,3H).MS(ESI):m / z 601.7[M+H] + .

[0452] Example 25: N-(3-(benzo[b]thiophene-2-carboxamido)-4-methylphenyl)-3,4-dihydro-2H-benzo[b][1,4]dioxepine-7-carboxamide (Example 25) [ka]

[0453] 4-Methylbenzene-1,3-diamine (30 mg, 0.246 mmol), 3,4-dihydro-2H-benzo[b][1,4]dioxepin-7-carboxylic acid (47.7 mg, 0.246 mmol) and NMI (59 μL, 0.737 mmol) were dissolved in dry DMF (2 mL). TCFH (82.7 mg, 0.295 mmol) was added in portions and the mixture was stirred at room temperature for 1 h. Benzo[b]thiophene-2-carboxylic acid (43.8 mg, 0.246 mmol) was added along with additional TCHF (82.7 mg, 0.295 mmol) and NMI (82.7 mg, 0.295 mmol) and the mixture was stirred at room temperature for 2 h. The mixture was concentrated to dryness and the residue was purified by flash column chromatography (gradient cyclohexane / EtOAc 3:1) to give the title compound as a white powder. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.20(s,1H),10.11(s,1H),8.32(s,1H),8.07(d,J=7.9Hz,1H),8.01(d,J=7.6Hz,1H),7.87(s,1H),7.62(d,J=6.6Hz,2H),7.58(s,1H),7.50 -7.48(m,2H),7.25(d,J=8.2,1H),7.07(d,J=8.2Hz,1H),4.24-4.19(m,4H),2.24(s,3H),2.20-2.15(m,2H).MS(ESI):m / z 459.5[M+H] + .

[0454] Example 26: N-(3-(6-(3-(3-hydroxyazetidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 26) [ka]

[0455] N-(3-(6-(3-(3-methoxyazetidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)-4-methylphenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 3 / 23) (30.0 mg, 0.05 mmol) was dissolved in dry DCM (2 mL) and the mixture was stirred at -78°C. Boron tribromide (204 μL, 0.20 mmol) was added dropwise. The dry ice / acetone bath was removed and the mixture was stirred at room temperature for 3 h. The reaction was quenched with water. The mixture was extracted with EtOAc (3 x 25 mL). The combined organic layers were concentrated to dryness and the residue was purified by preparative HPLC (20-40% MeCN in water) to give the title compound. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.04(s,2H),8.21(s,1H),7.86(s,1H),7.83(s,1H),7.59(s,1H),7.56(s,1H),7.52( s,1H),7.49(s,1H),7.22(d,J=8.3Hz,1H),7.05(d,J=8.8Hz,1H),6.97(d,J=8.3Hz,1H ),4.34-4.27(m,4H),4.20-4.12(m,1H),4.07(t,J=6.2Hz,2H),3.54(t,J=6.3Hz,2H), 2.71(t,J=6.2Hz,2H),2.57-2.53(m,2H),2.22(s,3H),1.80-1.71(m,2H).MS(ESI):m / z 574.7[M+H] + .

[0456] Example 27: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-((4-isopropylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 27) [ka]

[0457] Step 1: tert-Butyl-4-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)thieno[2,3-b]pyridin-6-yl)methyl)piperazine-1-carboxylate (27a) 6-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23) (21 mg, 0.056 mmol), N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (16 mg, 0.056 mmol) and NMI (13 μL, 0.167 mmol) were dissolved in dry DMF (2 mL). TCFH (18.7 mg, 0.067 mmol) was added portionwise and the mixture was stirred at room temperature for 1 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound.

[0458] Step 2: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-(piperazin-1-ylmethyl)thieno[2,3-b]pyridine-2-carboxamide (27b) tert-Butyl-4-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)thieno[2,3-b]pyridin-6-yl)methyl)piperazine-1-carboxylate (27a) (36 mg, 0.056 mmol) was dissolved in DCM (2 mL) and stirred at 0° C. TFA was added to the mixture until the ratio of DCM / TFA was 2:1. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated to dryness to give the title compound.

[0459] Step 3: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)-6-((4-isopropylpiperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 27) N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)-6-(piperazin-1-ylmethyl)thieno[2,3-b]pyridine-2-carboxamide (27b) (30 mg, 0.055 mmol), cesium carbonate (35.7 mg, 0.110 mmol) and 2-bromopropane (6 μL, 0.06 mmol) were dissolved in dry DMF (2 mL) and the mixture was stirred at room temperature overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.51(s,1H),10.16(s,1H),8.38(d,J=8.4Hz,1H),8.32(s,1H),8.10(d,J= 3.5Hz,1H),7.68-7.63(m,1H),7.59(d,J=8.4Hz,1H),7.53(s,1H),7.50(s, 1H),7.31(t,J=9.2Hz,1H),6.99(d,J=8.4Hz,1H),4.34-4.28(m,4H),3.71( s,2H),2.66-2.56(m,1H),2.46(s,8H),0.96(d,J=6.5Hz,6H).MS(ESI):m / z 590.6[M+H] + .

[0460] Example 27 / 1: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-(((1R,4R)-5-ethyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 27 / 1) [ka]

[0461] The title compound was prepared similarly as described for Example 27, using 6-(((1R,4R)-5-(tert-butoxycarbonyl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23 / 1) instead of 6-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23) in step 1, and ethyl iodide instead of 2-bromopropane in step 3. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.51(s,1H),10.16(s,1H),8.38(d,J=8.3Hz,1H),8.32(s,1H),8.21(s,1H),8.10(d,J=6.2Hz,1H),7.68-7 .61(m,2H),7.53(s,1H),7.50(s,1H),7.29(t,J=9.4Hz,1H),6.99(d,J=8.3Hz,1H),4.34-4.28(m,4H),3.99 -3.83(m,2H),3.42(s,1H),2.81-2.61(m,6H),1.74-1.64(m,2H)1.01(t,J=7.1Hz,3H).MS(ESI):m / z 588.6[M+H] + .

[0462] Example 27 / 2: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide)-2-fluorophenyl)-6-(((1S,4S)-5-ethyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 27 / 2) [ka]

[0463] The title compound was prepared similarly as described for Example 27 using 6-(((1S,4S)-5-(tert-butoxycarbonyl)-2,5-diazabicyclo[2.2.1]heptan-2-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23 / 2) instead of Int23 in step 1, and ethyl iodide instead of 2-bromopropane in step 3. 1 H NMR (300MHz, DMSO-d 6 ):δ ppm 10.51(s,1H),10.16(s,1H),8.38(d,J=8.3Hz,1H),8.32(s,1H),8.21(s,1H),8.10(d,J=6.2Hz,1H ),7.68-7.61(m,1H),7.53(s,1H),7.50(s,1H),7.29(t,J=9.4Hz,1H),6.99(d,J=8.3Hz,1H),4.34 -4.27(m,4H),3.99-3.83(m,2H),3.42(s,1H),2.81-2.61(m,6H),1.74-1.64(m,2H),1.01(t,J=7.1Hz,3H).MS(ESI):m / z 588.5[M+H] + .

[0464] Example 27 / 3: N-(5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)-6-((4-ethyl-1,4-diazepan-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxamide (Example 27 / 3) [ka]

[0465] The title compound was prepared similarly as described for Example 27, using 6-((4-(tert-butoxycarbonyl)-1,4-diazepan-1-yl)methyl)thieno[2,3-b]pyridine-2-carboxylic acid (Int23 / 3) instead of Int23 in step 1, and ethyl iodide instead of 2-bromopropane in step 3. 1H NMR (300MHz, DMSO-d 6 ):δ ppm 10.51(s,1H),10.16(s,1H),8.39(d,J=8.2Hz,1H),8.32(s,1H),8.17(s,1H),8.11(d,J=6.5Hz,1H),7.68-7.60(m,1H),),7.5 3(s,1H),7.49(s,1H),7.29(t,J=9.6Hz,1H),6.99(d,J=8.2Hz,1H),4.35-4.27(m,4H),3.89(s,2H),2.83-2.60(m,10H),1.83 -1.76(m,2H),1.03(t,J=7.1Hz,3H).MS(ESI):m / z 590.6[M+H] + .

[0466] Example 28: N-(4-fluoro-3-(6-(((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 28) [ka]

[0467] Step 1: 5-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)methyl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (3aR,6aS)-tert-butyl (28a) 6-(((3aR,6aS)-5-(tert-butoxycarbonyl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-yl)methyl)benzo[b]thiophene-2-carboxylic acid (Int23 / 4) (22.5 mg, 0.056 mmol), N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (16.1 mg, 0.056 mmol) and NMI (13 μL, 0.168 mmol) were dissolved in dry DMF (2 mL). TCFH (18.7 mg, 0.067 mmol) was added portionwise and the mixture was stirred at room temperature for 1 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound.

[0468] Step 2: N-(4-fluoro-3-(6-(((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 28) 5-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)methyl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (3aR,6aS)-tert-butyl (28a) (37.5 mg, 0.056 mmol) was dissolved in DCM and stirred at 0° C. To the mixture was added TFA until the ratio was DCM:TFA=2:1. The mixture was then stirred at rt for 2 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound. 1 H NMR (300MHz, DMSO-d 6):δ ppm 10.39(s,1H),10.15(s,1H),8.33(s,1H),8.10(d,J=7.1Hz,1H),7.94(s,1H) ,7.92(s,1H),7.69-7.62(m,1H),7.53(s,1H),7.49(s,1H),7.42(d,J=8.0Hz ,1H),7.29(t,J=9.5Hz,1H),6.98(d,J=8.4Hz,1H),4.35-4.27(m,4H),3.64( s,2H),2.80-2.71(m,2H),2.60-2.51(m,6H),2.32-2.21(m,2H).MS(ESI):m / z 573.5[M+H] + .

[0469] Example 29: N-(3-(6-((1-ethylpiperidin-4-yl)(hydroxy)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 29) [ka]

[0470] Step 1: 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid (29a) To a mixture of 6-bromobenzo[b]thiophene-2-carboxylic acid (100 mg, 0.389 mmol) in dry THF (2 mL) was added methylmagnesium bromide (Et 2 3M solution in hexanes (0.130 mL, 0.39 mmol) was added. The mixture was stirred at rt for 30 min and then cooled to -78 °C. To the mixture was added n-BuLi (2.5 M solution in hexanes (0.31 mL, 0.78 mmol) and the mixture was stirred at -78 °C for 1 h. Then tert-butyl 4-formylpiperidine-1-carboxylate (91.3 mg, 0.428 mmol) was added and the mixture was stirred at 0 °C overnight. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound.

[0471] Step 2: tert-Butyl 4-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)(hydroxy)methyl)piperidine-1-carboxylate (29b) Int2 (14.7 mg, 0.051 mmol), 6-((1-(tert-butoxycarbonyl)piperidin-4-yl)(hydroxy)methyl)benzo[b]thiophene-2-carboxylic acid (29a) (20.0 mg, 0.051 mmol) and NMI (14 μL, 0.179 mmol) were dissolved in dry DMF (2 mL). TCFH (14.3 mg, 0.051 mmol) was then added portionwise and the mixture was stirred at room temperature for 1 h. The mixture was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound as a white powder.

[0472] Step 3: N-(4-fluoro-3-(6-(hydroxy(piperidin-4-yl)methyl)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (29c) To a mixture of tert-butyl 4-((2-((5-(2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamido)-2-fluorophenyl)carbamoyl)benzo[b]thiophen-6-yl)(hydroxy)methyl)piperidine-1-carboxylate (29b) (33.7 mg, 0.051 mmol) in dry DCM (510 μL) was added TFA (100 μL) dropwise at 0° C. The mixture was allowed to warm to rt. Saturated NaHCO 3 (10 mL) was added to the mixture. The aqueous layer was extracted with DCM (3×15 mL). The combined organic layers were washed with MgSO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound.

[0473] Step 4: N-(3-(6-((1-ethylpiperidin-4-yl)(hydroxy)methyl)benzo[b]thiophene-2-carboxamide)-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Example 29) To a mixture of N-(4-fluoro-3-(6-(hydroxy(piperidin-4-yl)methyl)benzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (29c) (10.0 mg, 0.018 mmol) in dry DMF (1 mL) was added sodium cyanoborohydride (1.1 mg, 0.018 mmol). The mixture was cooled to 0° C. Acetaldehyde (1 μL, 0.018 mmol) was added and the mixture was stirred at 0° C. for 15 min. NaHCO 3 (2 mL) was added and the mixture was extracted with EtOAc (10 mL). The organic layer was concentrated to dryness and the residue was purified by preparative HPLC to give the title compound. 1 H NMR (300MHz, CD 3 OD):δ ppm 8.17 -8.13(m,2H),7.97-7.94(m,2H),7.61-7.55(m,1H),7.50-7.45(m,3H),7.23 (t,J=9.6Hz,1H),6.96(d,J=8.3Hz,1H),4.58-4.54(m,2H),4.36-3.29(m,4H ),3.22-3.14(m,1H),2.76-2.66(m,2H),2.41-2.25(m,2H),2.11-2.06(m,1H ),1.91-1.79(m,1H),1.62-1.44(m,3H),1.19(t,J=7.1Hz,3H).MS(ESI):m / z 590.6[M+H] +

[0474] Example 30: N-(4-fluoro-3-(6-(2-hydroxy-3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (racemic) (Example 30) [ka]

[0475] Step 1: N-(4-fluoro-3-(6-hydroxybenzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30a) To a mixture of 6-hydroxybenzo[b]thiophene-2-carboxylic acid (2.0 g, 10.3 mmol), N-(3-amino-4-fluorophenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (Int2) (800 mg, 2.8 mmol) and TEA (28 mg, 0.28 mmol) in DMF (50 mL) was added HATU (3.0 g, 7.9 mmol) and the mixture was stirred at 50° C. for 2 h. To the mixture was added KOH (313 mg, 5.6 mmol) as an aqueous solution. The mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (3×100 mL) and concentrated with Na 2 SO 4 It was dried over water, filtered and concentrated to dryness. The residue was purified by column chromatography on silica gel (PE / EtOAc=10:1) to give the title compound as a brown solid.

[0476] Step 2: N-(4-fluoro-3-(6-(oxiran-2-ylmethoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30b) A mixture of N-(4-fluoro-3-(6-hydroxybenzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30a) (210 mg, 0.45 mmol) in DMF (10 mL) was cooled to 0° C. NaH (22 mg, 0.91 mmol) was added and the mixture was stirred at 0° C. for 0.5 h. 2-(Chloromethyl)oxirane (54 mg, 0.58 mmol) was added and the mixture was stirred at 60° C. for 4 h. Water (100 mL) was added and the mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were washed with brine (3×100 mL) and diluted with Na 2 SO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0477] Step 3: N-(4-fluoro-3-(6-(2-hydroxy-3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (racemic) (Example 30) To a mixture of N-(4-fluoro-3-(6-(oxiran-2-ylmethoxy)benzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30b) (210 mg, 0.40 mmol), TEA (40 mg, 40 mmol) in ethanol (5 mL) was added pyrrolidine (142 mg, 2.00 mmol). The mixture was stirred at 80° C. under microwave irradiation for 2 h. The mixture was concentrated to dryness and the residue was purified by column chromatography on silica gel (DCM / MeOH=15:1) to give the crude product as a white solid. The crude product was purified by preparative HPLC to give the title compound as a white solid. 1 H NMR (400MHz, DMSO-d 6):δ ppm 10.34(s,1H),10.17(s,1H),8.27(s,1H),8.10-8.08(m,1H),7.88(d,J=8.9Hz,1H),7.68-7.61(m ,2H),7.55-7.48(m,2H),7.31-7.26(m,1H),7.11-7.08(m,1H),6.99(d,J=8.4Hz,1H),4.95(broad s,1H),4.34-4.29(m,4H),4.11-4.07(m,1H),4.00-3.94(m,2H),2.70-2.63(m,1H),2.53-2.46(m,5H),1.70-1.67(m,4H).MS(ESI):m / z 592.2[M+H] + .

[0478] Example 30 / 1: N-(4-fluoro-3-(6-(2-hydroxy-3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (single enantiomer) (Example 30 / 1) [ka]

[0479] Step 1: N-(4-fluoro-3-(6-(oxiran-2-ylmethoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30 / 1a) To a solution of N-(4-fluoro-3-(6-hydroxybenzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30a) (1.0 g, 2.16 mmol) in DMF (10 mL) was added NaH (172 mg, 60% dispersion in mineral oil, 4.3 mmol) under nitrogen atmosphere at 0° C. (S)-2-(chloromethyl)oxirane (297 mg, 3.21 mmol) was added to the mixture. The mixture was stirred at 60° C. for 1.5 h. Water was added (100 mL) and the mixture was extracted with DCM (2×100 mL). The combined organic layers were washed with Na2 SO 4 It was dried over ice, filtered and concentrated to dryness to give the title compound, which was used in the next step without further purification.

[0480] Step 2: N-(4-fluoro-3-(6-(2-hydroxy-3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (single enantiomer) (30 / 1) A solution of N-(4-fluoro-3-(6-(oxiran-2-ylmethoxy)benzo[b]thiophene-2-carboxamido)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (30 / 1a) (400 mg, 0.77 mmol) and pyrrolidine (56 mg, 0.79 mmol) in EtOH (10 mL) was stirred at 70° C. overnight. The mixture was concentrated to dryness and the residue was purified by silica gel chromatography (DCM / MeOH=20:1) to give the crude product, which was purified by reverse phase column (CH 3 CN / H 2 O = 5%-95%), followed by chiral SFC (column: CHIRALCEL OD-3, 4.6 * 100 mm, co-solvent: MeOH / acetonitrile (0.2% 7M NH 3 )) to afford the title compound as an off-white solid (second eluting enantiomer, retention time = 1.9 min). 1 H NMR (400MHz, DMSO-d 6):δ ppm 10.36(s,1H),10.18(s,1H),8.27(s,1H),8.11-8.09(m,1H),7.88(d,J=8Hz,1H),7.67-7.63(m,2H),7.54-7.50(m,2H),7.31-7.2 7(m,1H),7.11-7.08(m,1H),7.00(d,J=8Hz,1H),4.96-4.95(m,1H),4.33-4.30(m,4H),4.11-4.08(m,1H),3.99-3.95(m,2H),2.66 -2.62(m,1H),2.50-2.43(m,5H),1.71-1.65(m,4H).MS(ESI):m / z 592.3[M+H] + .

[0481] Example 30 / 2: N-(4-fluoro-3-(6-(2-hydroxy-3-(pyrrolidin-1-yl)propoxy)benzo[b]thiophene-2-carboxamide)phenyl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (single enantiomer) (Example 30 / 2) [ka]

[0482] The title compound was prepared similarly as described for Example 30 / 1, using (R)-2-(chloromethyl)oxirane instead of (S)-2-(chloromethyl)oxirane in step 1. Chiral SFC (column: CHIRALCEL OD-3 (4.6 * 100 mm), co-solvent: MeOH / acetonitrile (0.2% 7M NH 3 Final purification by HPLC afforded the title compound as an off-white solid (first eluting enantiomer, retention time = 1.5 min). 1 H NMR (400MHz, DMSO-d 6):δ ppm 10.35(s,1H),10.18(s,1H),8.27(s,1H),8.11-8.09(m,1H),7.88(d,J=8Hz,1H), 7.67-7.63(m,2H),7.54-7.50(m,2H),7.31-7.26(m,1H),7.11-7.08(m,1H),7.00 (d,J=8Hz,1H),4.95-4.94(m,1H),4.33-4.30(m,4H),4.12-4.08(m,1H),3.99-3. 95(m,2H),2.67-2.62(m,1H),2.48-2.43(m,5H),1.71-1.64(m,4H).MS(ESI):m / z 592.3[M+H] + .

[0483] Biological assays Assay for inhibition of ATP production The assay determines the amount of ATP in the cells in culture.

[0484] Cells were plated at 4,000 per well in 96-well plates in high glucose DMEM (PAN) supplemented with 10% FBS (PAN), 2 mM glutamine-alanine (PAN), 2 mM pyruvate (Sigma) and 10 mL / L penicillin-streptomycin and incubated with 0.5% O 2 , 5% CO 2 For hypoxia, cells were grown at 37°C in 0.5% O and 75% humidity in a Whitley H35 Hypoxystation (Don Whitley Scientific). 2 , 5% CO 2The cells were grown and treated at 37°C with 5% CO and 75% humidity. The following day, the cells were induced with test compounds (Figure 1: compounds of Examples 3 / 16, 3 / 8, 10 at 10000, 3333, 1111, 370, 123, 41, 14, 5 and 1.5 nM; Figures 8A and 8B: compounds of Examples 3 / 19, 30 and 30 / 2 at 10000, 3333, 1111, 370, 123, 41, 14, 5 and 1.5 nM) and with vehicle (DMSO) alone for 48 hours. After this time, the medium was removed from the cells and intracellular ATP levels were determined using the Promega CellTiter-Glo2.0 assay protocol (Promega, G9241). Sample values ​​were compared to DMSO alone to estimate the effect of compounds on ATP production. IC 50 Values ​​were generated by plotting luciferase counts against compound concentration.

[0485] Figure 1 contains several graphs showing the inhibitory activity of the compounds of Examples 3 / 16, 3 / 8 and 10 in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cells after 48 hours of treatment. In all cases, a dose-dependent reduction in ATP production levels is observed.

[0486] Figures 8A and 8B show the inhibitory activity of compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 in Panc02 and SKOV3 cells after 48 hours of treatment. In all cases, a dose-dependent reduction in ATP production levels is observed. The numerical results are shown in Table 1 below, and the compound activity was evaluated using the following IC 50 Group by range: + 1μM-10μM, ++ 100nM-<1μM +++<100nM

[0487] Lactate production inhibition assay Recombinant lactate dehydrogenase A (LDHA) was used to +Lactate in tissue culture supernatants was converted to pyruvate by the simultaneous reduction of NAD to NADH. The electron acceptor 1-methoxyphenazine methosulfate (MPMS) was then reduced by the resulting NADH, which resulted in the conversion of NAD + The reduced 1-methoxyphenazine methosulfate is then used to reduce water-soluble tetrazolium 8 (WST8) to the orange WST8 formazan product. The assay is adapted from the article "Application of WST-8 based colorimetric NAD(P)H detection for quantitative dehydrogenase assays" in Chamchoy et al., BMC Biochemistry (2019) 20(1):4.

[0488] Cells were plated at 4,000 per well in 96-well plates in high glucose DMEM (PAN) supplemented with 10% FBS (PAN), 2 mM glutamine-alanine (PAN), 2 mM pyruvate (Sigma) and 10 mL / L penicillin-streptomycin (PAN) and 0.5% O 2 , 5% CO 2 For hypoxia, cells were grown at 37°C in 0.5% O and 75% humidity in a Whitley H35 Hypoxystation (Don Whitley Scientific). 2 , 5% CO 2The cells were grown and treated at 37° C. with 75% humidity and 5% CO. The following day, the cells were induced with test compounds (FIG. 2: compounds of Examples 3 / 16, 3 / 8, 10 at 10000, 3333, 1111, 370, 123, 41, 14, 5 and 1.5 nM; FIG. 7: compounds of Examples 3 / 16, 3 / 8, 10 at 41, 14, 4.6 nM and felformin at 100, 50, 25, 12.5, 6.25 μM; FIG. 8C and 8D: compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 at 10000, 3333, 1111, 370, 123, 41, 14, 5 and 1.5 nM) and vehicle (DMSO) alone for 48 hours. After this time, the medium was removed from the cells and extracellular lactate was measured. To detect extracellular lactate, cell supernatant was diluted 1:5 in assay buffer (0.2M Tris, pH 8.2) by adding 10 μl cell supernatant + 40 μl buffer in a 96-well plate. Assay substrate mix was prepared by mixing NAD+ (3.75 mM), WST-8 (0.9 mM), MPMS (10 mM), and recombinant LDHA expressed in E. coli as an N-terminal hexahistidine tagged fusion protein was added to a final concentration of 0.8 μg / ml in assay buffer. Then, 80 μl of the substrate mix was added to 20 μl of diluted cell supernatant in a 96-well flat-bottom plate, and the absorbance at 460 nm was measured at 1-minute intervals over a period of 30 minutes. Standard curves of known lactate concentrations of 10, 8, 6, 4, 1, 0.5 and 0.25 mM were constructed for each assay plate.

[0489] Figure 2 shows the downregulation of lactate levels in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cells after 48 hours of treatment with compounds of Examples 3 / 16, 3 / 8 and 10. In all cases, a dose-dependent reduction in lactate production levels is observed.

[0490] FIG. 7 shows the downregulation of lactate levels in HeLa cells following 48 hour co-treatment of ferformin with compounds of Examples 3 / 16, 3 / 8, and 10.

[0491] Figures 8C and 8D show dose-dependent downregulation of lactate levels in Panc02 and SKOV3 cells after 48 hours treatment with compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2.

[0492] The numerical results are shown in Table 1 below, and the compound activity was evaluated using the following IC 50 Group by range: + 1μM-10μM, ++ 100nM-<1000nM +++<100nM

[0493] Table 1 [Table 5-1] [Table 5-2] [Table 5-3]

[0494] As can be seen in the above table, all tested compounds of the present invention showed activity in inhibiting the production of ATP and / or lactate. In particular, compounds of the present invention of Examples 10, 3 / 8, 3 / 16, 3 / 19, 30 and 30 / 2 demonstrated high activity in inhibiting either or both of ATP and lactate.

[0495] Gene expression analysis by qPCR Cells were plated at 40,000 cells / ml in high glucose DMEM (PAN) supplemented with 10% FBS (PAN), 2 mM glutamine-alanine (PAN), 2 mM pyruvate (Sigma) and 10 mL / L penicillin-streptomycin (PAN) and 0.5% O 2 , 5% CO 2The cells were grown at 37°C with 5% CO and 75% humidity. The next day, the cells were induced with test compounds (Figure 3: compounds of Examples 3 / 16, 3 / 8, 10 at 10 nM; Figures 8E-8AB: compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2 at 10 nM) for 0.5, 1, 2 and 4 hours, as well as with vehicle (DMSO) alone. After this time, the cells were lysed according to a protocol (Promega, SV 96 Total RNA Isolation System, no. Z3505). After transferring the lysates to a 96-well RNA binding plate, RNA was isolated and approximately 1 μg was subjected to cDNA synthesis using the LunaScript RT SuperMix Kit (NEB, no. E3010). The resulting cDNA was diluted 1:5 and used for qPCR analysis (Taqman). Luna Universal Probe qPCR Master Mix (NEB, no. M3004) is used as the 2x reaction mix to which the diluted cDNA is added. qPCR analysis was performed using the QuantStudio™ 6Flex Real-Time PCR System (Thermo Fisher Scientific).

[0496] A set of primers and a probe for each gene of interest were purchased from IDT (Integrated DNA Technologies). The corresponding sequences are summarized in Table 2:

[0497] Table 2: [Table 6-1] [Table 6-2]

[0498] FIG. 3 shows the time-dependent upregulation of ER stress markers CHAC, CHOP / DDIT3, XBP1 and SLC7A11 in HeLa, 4T1, LLC1, A549, MDA-MB-231 and MiaPaCa2 cells induced with the compounds of Examples 3 / 16, 3 / 8 and 10.

[0499] Figures 8E-8AB show the time-dependent upregulation of ER stress markers CHAC and CHOP / DDIT3 in Panc02 and SKOV3 cells induced with compounds of Examples 3 / 16, 3 / 8, 10, 3 / 19, 30 and 30 / 2.

[0500] Western blot analysis Western blot analysis was used to detect the amount of a particular protein in differently treated cells. Proteins were separated by size, transferred onto nitrocellulose membranes, and incubated with protein-specific antibodies. The density of the observed bands is proportional to the amount of a particular protein in the cells.

[0501] The cells were then placed in a 10 cm 2 4 x 10 per plate 6 (for hypoxia) or 3×10 6 (For normoxia) Cells were cultured in high glucose DMEM (PAN) supplemented with 10% FBS (PAN), 2 mM glutamine-alanine (PAN), 2 mM pyruvate (Sigma) and 10 ml / L penicillin-streptomycin (PAN) and maintained at 0.5% O 2 , 5% CO 2 For hypoxia, cells were grown at 37°C in 0.5% O and 75% humidity in a Whitley H35 Hypoxystation (Don Whitley Scientific). 2 , 5% CO 2Cells were grown and treated at 37°C with 75% humidity and 5% CO. The next day, the cells were induced with test compounds (Figure 4: Compounds of Examples 3 / 8 and 10 at concentrations of 0.1, 0.3 and 1 μM, Bafilomycin A at 20 nM; Figure 5: Compounds of Examples 3 / 16, 3 / 8 and 10 at concentrations of 10, 30 and 100 nM, Bafilomycin A at 20 nM) and vehicle (DMSO) alone for 24 hours. After 24 hours, the cells were washed with PBS and harvested by scraping. Lysates were centrifuged at 2,000 rpm for 3 minutes at 4°C. The cell pellet was removed by lysis in 250 μl lysis buffer (20 mM Tris pH 7.8, 150 mM NaCl, 0.05% Tween 20, 2 mM MgCl2, protease inhibitors (Roche), 1 mM NaF, 1 mM Na 3 VO 4) and sonicated at 50% for 20 seconds. Samples were centrifuged at 14,000 rpm for 4 minutes at 4°C. Protein concentrations of the samples were measured using a DC protein assay kit (Biorad, 5000112) and 22 μg of protein extract per well was loaded onto a precast SDS page gel (Biorad, 1704273). Proteins were transferred onto a PVDF-membrane (Biorad, 10026933) using the Biorad Transblot Turbo transfer system. The membrane was then incubated in blocking buffer (5% dry milk, 0.1% Tween 20, Tris-buffered saline buffer) for 45 minutes, followed by primary antibodies against HIF1a (no. 36169, Cell Signaling) at a dilution of 1:1000, vinculin (7F9) (no. sc-73614, Santa Cruz Biotechnology) at a dilution of 1:10000, SQSTM1 / p62(D-3) (no. sc-28359, Santa Cruz Biotechnology) at a dilution of 1:1000, p70 S6 kinase (no. 9202, Cell Signaling) at a dilution of 1:2000, and phospho-p70 S6 kinase (Thr389) (no. 9205, Cell Signaling) at a dilution of 1:1000 in the blocking buffer overnight. The next day, the membrane was washed three times for 10 min with 0.1% Tween20 in TBS buffer and incubated with secondary antibody anti-rabbit IgG HRP (no. sc-2004, Santa Cruz Biotechnology) or anti-mouse IgG HRP (no. sc-2005, Santa Cruz Biotechnology) in the blocking buffer for 45 min. The membrane was then washed three times for 10 min. Chemiluminescence was measured using a chemiluminescent substrate (no. 34580, Thermo Scientific).

[0502] FIG. 4 shows the dose-dependent reduction of p62 and HIF1α protein levels in HeLa cells treated with compounds of Examples 3 / 8 and 10 for 24 hours under normoxia and hypoxia.

[0503] FIG. 5 shows dose-dependent reduction of p62 in HeLa cells treated for 24 hours with compounds of Examples 3 / 16, 3 / 8 and 10, also in cells simultaneously treated with bafilomycin A under normoxia and hypoxia.

[0504] FIG. 5 also shows the dose-dependent induction of p70-S6K phosphorylation (Thr389) in HeLa cells treated for 24 hours with the compounds of Examples 3 / 16, 3 / 8 and 10, and in cells simultaneously treated with bafilomycin A under normoxia.

[0505] Determination of autophagy flux Autophagy flux was determined using a HeLa cell line stably expressing an artificial protein construct consisting of green fluorescent protein (GFP) as an N-terminal fusion to human full-length LC3 protein preceded by red fluorescent protein (RFP) as an N-terminal fusion to human LC3 lacking a C-terminal glycine residue (LC3DG) (Kaizuka et al., Mol. Cell (2016) 17 835-849). This construct is cleaved intracellularly by ATG4 at the C-terminus of the full-length LC3 protein, where the GFP-LC3 fragment becomes conjugated to phosphatidylethanolamine. RFP-LC3DG cannot be lipidated and can be used as an internal reference to generate ratiometric values ​​reporting autophagy flux. Lipidated GFP-LC3 protein becomes incorporated into the membrane of autophagosomes and degrades upon fusion with lysosomes. As a result, cells with high autophagic flux have little GFP-LC3, whereas low autophagic flux results in relatively high intracellular GFP-LC3 amounts.

[0506] Reporter cells were grown as subconfluent cultures and test compounds were applied at various concentrations. Thapsigargin, a known inducer of autophagy, was used as a positive control. After 24 h of treatment, single-cell suspensions were prepared by trypsinization and GFP fluorescence was determined by flow cytometry. A minimum of 10,000 single cells were assessed for each condition.

[0507] FIG. 6 shows that the tested compounds of Examples 3 / 8, 3 / 16 and 10 significantly enhance autophagic flux in a HeLa cell line expressing an artificial protein construct consisting of GFP-LC3.

Claims

1. A compound of formula (I) or an enantiomer, diastereomer, N-oxide, solvate or pharma- ceutically acceptable salt thereof: 【Chemistry 1】 During the ceremony, K is O, S or C(R ５ ) ２ and L is O, S or C(R ５ ) ２ and n is 0, 1, 2, 3 or 4; p is 1 or 2; q are independently 0, 1 or 2; r is 1 or 2; A is C １－６ Alkyl, halogen, CN, halo-C １－６ Alkyl, C ３－６ Cycloalkyl, halo-C ３－６ Cycloalkyl, OH, —OC １－６ Alkyl or O-halo-C １－６ is alkyl; X is S, O, NR ６ , N or CR ７ and Y is S, O, NR ６ , N or CR ７ and Z is C or N; D represents a 6-membered aryl or a 5-6 membered heteroaryl containing 1-4 heteroatoms independently selected from O, N, and S; R １ , F, C １－３ Alkyl or spirocyclic fused C ３－６ is cycloalkyl; R ２ are independently halogen or C １－６ is alkyl; R ３ is hydrogen, halogen, C １－６ Alkyl, halo-C １－６ Alkyl, C ３－６ Cycloalkyl, halo-C ３－６ Cycloalkyl, -OC １－６ Alkyl or O-halo-C １－６ is alkyl; R ４ is hydrogen, halogen or V １ -V ２ -V ３ -V ４ -V ５ and Where: V １ is absent, a bond, or C １－６ is alkylene; V ２ is absent, a bond, or O, S, S(O) ｘ , S(O)(=NR ａ ), S(O) ２ N.R. ａ , N.R. ａ , N.R. ａ C(O) or C(O)NR ａ and V ３ is absent, a bond, or C １－６ is alkylene; Here, V １ Or V ３ C in １－６ Alkylene is unsubstituted or can be selected from the group consisting of OH, C １－６ Alkyl, C １－６ Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen and oxo; or V １ Or V ３ C in １－６ Two C's on the alkylene １－６ The alkyl groups, together with the carbon atom to which they are attached, are C ３－６ forming a cycloalkyl group; V ４ is absent, a bond, or O, S, S(O) ｘ , S(O)(=NR ａ ), S(O) ２ N.R. ａ , N.R. ａ , N.R. ａ C(O) or C(O)NR ａ and V ５ is hydrogen, halogen, OH, CN, CO ２ H, CO ２ -C １－６ Alkyl, N(R ａ ) ２ , C １－６ Alkyl, C ３－８ cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, phenyl, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl, phenyl or heteroaryl is unsubstituted or substituted with CN, CO ２ H, CO ２ -C １－６ Alkyl, N(R ａ ) ２ , O.H., C. １－６ Alkyl, O-C １－６ Alkyl, C(O)-C １－６ Alkyl, C １－６ Haloalkyl, halogen, oxo, spiro fused ring C ３－６ cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; Ｖ５ is replaced by; R ５ is hydrogen, F, C １－３ Alkyl or spirocyclic fused C ３－６ is cycloalkyl; R ６ is hydrogen or C １－６ is alkyl; R ７ is hydrogen, halogen or C １－６ is alkyl; R ａ is hydrogen or C １－３ is alkyl; X is 1 or 2.

2. 2. The compound of claim 1 represented by formula (I), or an enantiomer, diastereomer, N-oxide, solvate, or pharma- ceutically acceptable salt thereof: 【Chemistry 2】 During the ceremony, K is O, S or C(R ５ ) ２ and L is O, S or C(R ５ ) ２ and n is 0, 1, 2, 3 or 4; p is 1 or 2; q are independently 0, 1 or 2; r is 1 or 2; A is C １－６ Alkyl, halogen, CN, halo-C １－６ Alkyl, C ３－６ Cycloalkyl, halo-C ３－６ Cycloalkyl, OH, —OC １－６ Alkyl or O-halo-C １－６ is alkyl; X is S, O, NR ６ , N or CR ７ and Y is S, O, NR ６ , N or CR ７ and Z is C or N; D represents a 6-membered aryl or a 5-6 membered heteroaryl containing 1-4 heteroatoms independently selected from O, N, and S; R １ , F, C １－３ Alkyl or spirocyclic fused C ３－６ is cycloalkyl; R ２ are independently halogen or C １－６ is alkyl; R ３ is hydrogen, halogen, C １－６ Alkyl, halo-C １－６ Alkyl, C ３－６ Cycloalkyl, halo-C ３－６ Cycloalkyl, -OC １－６ Alkyl or O-halo-C １－６ is alkyl; R ４ is hydrogen, halogen or V １ -V ２ -V ３ -V ４ -V ５ and Where: V １ is absent, a bond, or C １－６ is alkylene; V ２ is absent, a bond, or O, S, S(O) ｘ , S(O)(=NR ａ ), S(O) ２ N.R. ａ , N.R. ａ , N.R. ａ C(O) or C(O)NR ａ and V ３ is absent, a bond, or C １－６ is alkylene; Here, V １ Or V ３ C in １－６ Alkylene is unsubstituted or １－６ Alkyl, C １－６ Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen and oxo; or V １ Or V ３ C in １－６ Two C's on the alkylene １－６ The alkyl groups, together with the carbon atom to which they are attached, are C ３－６ forming a cycloalkyl group; V ４ is absent, a bond, or O, S, S(O) ｘ , S(O)(=NR ａ ), S(O) ２ N.R. ａ , N.R. ａ , N.R. ａ C(O) or C(O)NR ａ and V ５ is hydrogen, halogen, OH, CN, CO ２ H, CO ２ -C １－６ Alkyl, N(R ａ ) ２ , C １－６ Alkyl, C ３－８ cycloalkyl, 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, phenyl, or 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl, phenyl or heteroaryl is unsubstituted or substituted with CN, CO ２ H, CO ２ -C １－６ Alkyl, N(R ａ ) ２ , O.H., C. １－６ Alkyl, O-C １－６ Alkyl, C(O)-C １－６ Alkyl, C １－６ Haloalkyl, halogen, oxo, spiro fused ring C ３－６ cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; Ｖ５ is replaced by; R ５ is hydrogen, F, C １－３ Alkyl or spirocyclic fused C ３－６ is cycloalkyl; R ６ is hydrogen or C １－６ is alkyl; R ７ is hydrogen, halogen or C １－６ is alkyl; R ａ is hydrogen or C １－３ is alkyl; X is 1 or 2.

3. The compound is represented by formula (II): 【Chemistry 3】 U １ , U ２ , U ３ and U ４ The compound according to claim 1 or 2, wherein is independently selected from the group consisting of N and C.

4. The compound is represented by formula (III): 【Chemistry 4】 During the ceremony U １ and U ４ is independently selected from N and C; X is S, O, NR ６ or N; Y is CR ７ or N. The compound according to any one of claims 1 to 3.

5. The compound according to any one of claims 1 to 4, wherein n is 0.

6. The compound according to any one of claims 1 to 5, wherein p is 1.

7. The compound is represented by formula (IV): 【Chemistry 5】 During the ceremony, A is C １－６ is alkyl or halogen; The compound according to any one of claims 1 to 6, wherein X is S, O or NH.

8. R ４ is hydrogen, halogen or V １ -V ２ -V ３ -V ４ -V ５ where: V １ is absent, a bond, or C １－４ is alkylene; V ２ is absent, a bond, or O, S, S(O) ｘ , N.R. ａ and V ３ is absent, a bond, or C １－４ is alkylene, V ４ is absent, a bond, or O, S, S(O) ｘ , N.R. ａ and Here, V １ Or V ３ C in １－４ Alkylene is unsubstituted or １－３ Alkyl, C １－３ Substituted by 1 to 4 substituents independently selected from the group consisting of haloalkyl, halogen and oxo; or V １ Or V ３ C in １－４ Two C's on the alkylene １－６ The alkyl groups, together with the carbon atom to which they are attached, are C ３－６ forming a cycloalkyl group; V ５ is hydrogen, halogen, OH, CN, C １－６ Alkyl, C ３－６ cycloalkyl, a 4-8 membered monocyclic or bicyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N, or a 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms independently selected from the group consisting of O, S and N; wherein alkyl, cycloalkyl, heterocycloalkyl or heteroaryl is unsubstituted or is selected from the group consisting of OH, C １－６ Alkyl, O-C １－６ Alkyl, C(O)-C １－６ Alkyl, C １－６ Haloalkyl, halogen, oxo, spiro fused ring C ３－６ cycloalkyl, and one to four substituents R independently selected from the group consisting of spirocyclic fused 3- to 7-membered heterocycloalkyl containing one heteroatom selected from the group consisting of O, S, and N; Ｖ５ The compound according to any one of claims 1 to 7, substituted by:

9. The compound according to any one of claims 1 to 8, and N-oxides, solvates or pharma- ceutically acceptable salts thereof, selected from the group consisting of: 【Chemistry 6-1】 【Chemistry 6-2】 【Chemistry 6-3】 【Chemistry 6-4】 【Chemistry 6-5】 【Chemistry 6-6】 【Chemistry 6-7】

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

11. A compound according to any one of claims 1 to 9 for use as a medicine.

12. A compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 for use in a method for the prevention and / or treatment of a disease or condition mediated by the lactate / ATP mechanism.

13. 13. The compound or pharmaceutical composition of claim 12, wherein the disease or condition mediated by the lactate / ATP mechanism is cancer.

14. The compound or the pharmaceutical composition is - with one or more therapeutic agents for cancer selected from the group consisting of PD-1 agents, PD-L1 agents, CTLA-4 agents, IDO1 inhibitors, and anti-cancer vaccines; or - in conjunction with cytokine therapy or known chemotherapy or drug therapy, or during radiation therapy; 14. The compound or pharmaceutical composition of claim 13,

15. A compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 for use in a method for the prevention and / or treatment of a disease or condition, preferably cancer, by inducing endoplasmic reticulum stress in cells affected by said disease or condition, thereby reducing intracellular ATP and lactate production in said cells.

16. A compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 for use in a method for the prevention and / or treatment of cancer, preferably cancer cells and / or cancerous tumors, comprising The compound or pharmaceutical composition reduces Hif1-alpha at the protein level in hypoxic areas of the cancer, thereby downregulating ATP and lactate production in these areas and ultimately leading to a reduction in hypoxic areas and a reduction (preferably an efficient reduction) in the growth of the cancer.

17. A compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 for use in a method for the prevention and / or treatment of a disease or condition susceptible to the induction of endoplasmic reticulum stress, comprising The compound or pharmaceutical composition, wherein administration of the compound or pharmaceutical composition to a subject having or suspected of having the disease or condition results in a reduction in protein levels of Hif-1 alpha.

18. A compound according to any one of claims 1 to 9 or a pharmaceutical composition according to claim 10 for use in a method for the prevention and / or treatment of an autophagy-related disease selected from neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and Huntington's disease, amyotrophic lateral sclerosis, metabolic disorders such as obesity and diabetes mellitus type I and type II, viral infections and diseases leading to the accumulation of certain debris particles, such as fibrotic lung diseases.

19. A composition comprising a compound according to any one of claims 1 to 9 and a biguanide.

20. A kit comprising a compound according to any one of claims 1 to 9 and a biguanide.

21. 21. A composition according to claim 19 or a kit according to claim 20 for use as a medicament.

22. A combination of a compound according to any one of claims 1 to 9 and a biguanide for use in therapy.

23. A combination of a compound according to any one of claims 1 to 9 and a biguanide, or a kit according to claim 20, for use in a method for the prevention and / or treatment of cancer, preferably advanced stage cancer, preferably advanced stage glycolytic cancer.

24. 24. The combination or kit of claim 23, wherein the cancer is an LKB-1-deficient cancer.

25. The combination or kit of claim 23 or 24, wherein the cancer is an ARID1a-deficient cancer.

26. 26. The combination or kit of any one of claims 23 to 25, wherein the cancer is selected from the group consisting of non-small cell lung cancer, sarcoma, gynecological cancer such as cervical, ovarian cell or uterine carcinoma, adenoid cystic carcinoma, pancreatic cancer, other gastrointestinal cancer such as gastric cancer or esophageal cancer.

27. 20. The composition of claim 19, the kit of claim 20, the composition or kit of claim 21, the combination of claim 22, or the combination or kit of any one of claims 23 to 26, wherein the biguanide is selected from the group consisting of metformin, buformin and phenformin.

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