Therapeutic Compounds and Methods

Abstract

The present invention relates to a compound of formula (I): TIFF2024527604000134.tif18170 (in the formula, R 1 and R 2 has any of the values ​​described herein) or a prodrug thereof, or a pharma- ceutically acceptable salt thereof, and compositions comprising the compound of formula (I), or a prodrug thereof, or a pharma- ceutically acceptable salt thereof. The compounds are agonists of the glycolytic enzyme phosphofructokinase-1 hepatic form, and are useful in the treatment of diseases associated with the activity of the glycolytic enzyme phosphofructokinase-1 hepatic form, such as cancer, diabetes, sepsis, and septic shock.

Description

[Technical field]

[0001] Priority This application claims priority to U.S. Provisional Patent Application No. 63 / 222,288, filed July 15, 2021, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Phagocytes produce bactericidal reactive oxygen species (ROS) within the phagosome in an oxidative burst. The rapid increase in ROS is mediated by NOX2, a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent oxygen reductase (Thomas, DC, Immunol. Lett., 2017, 192, 88-96). The formation of NOX2 complexes on the phagosome and plasma membrane is accompanied by increased oxygen consumption and glucose uptake (Zatti, M., and Rossi, F. Biochim. Biophys. Acta, 1965, 99, 557-561). Glucose catabolism via the pentose phosphate pathway increases the production of NADPH, providing the reducing equivalents of NOX2 required to generate superoxide radicals.

[0003] Neutrophils are professional phagocytes essential for optimal antibacterial defense and constitute 50-70% of circulating leukocytes in humans (Mayadas, TN et al., Ann. Rev. Pathol., 2014, 9, 181-218). Neutrophils depend on the oxidative burst for numerous functions, including phagocytosis (Rosales, C, and Uribe-Querol, E., Biomed Res. Int., 2017, 9042851. doi:10.1155 / 2017 / 9042851), degranulation (Sengelov, H., et al., J. Immunol., 1995, 154, 4157-4165), ROS production (Amulic, B., et al., Annu. Rev. Immunol., 2012, 30, 459-489), and formation of neutrophil extracellular traps (NETs) (Brinkmann, V., J. Innate Immun., 2018, 10, 414-421; and Brinkmann, V., et al., Science, 2004, 303, 1532-1535). Mutations that inactivate the NOX2 complex impair the oxidative capacity of neutrophils and cause chronic granulomatous disease (CGD) (Roos, D., et al., Blood Cells Mol. Dis., 2010, 45, 246-265). Patients with CGD are prone to recurrent, chronic and invasive bacterial and fungal infections (Heyworth, PG, et al., Curr. Opin. Immunol., 2003, 15, 578-584).

[0004] Neutrophils are important for innate immunity, but excessive neutrophil activation can be harmful. Local tissue damage, inflammation and autoantigens caused by NETs exacerbate the pathology of chronic conditions such as atherosclerosis, psoriasis, gout and lupus (Brinkmann, V., J. Innate Immun., 2018, 10, 414-421). Targeting the oxidative burst may have therapeutic potential, but there are safety concerns regarding inhibitors of NOX2 or enzymes of the pentose phosphate pathway, including glucose-6-phosphate dehydrogenase (G6PDH). Barriers to their use include suppression of innate immunity and general toxicity (Diebold, BA, et al., Antioxid. Redox Signal, 2015, 23, 375-405; and Kowalik, MA, Columbano, A., and Perra, A., Oncol., 2017, 7, 87).

[0005] NETosis in neutrophils is crucial for killing extracellular bacteria (Brinkmann, V., et al., Science, 2004, 303, 1532-1535), but the underlying molecular mechanisms are largely unknown. Most physiological stimuli, including bacteria, fungi, and crystalline microparticles, trigger NOX2-dependent NETosis, whereas some bacterial toxins acting as potassium and calcium ionophores promote NOX2-independent NETosis (Kenny, EF, et al., Elife6, 2017, e24437). NOX2-dependent NETosis has been described as a two-step process (Neubert, E., et al., J. Cell Sci. 133, 2020, jcs241075). In step 1, an active signaling cascade triggers a NOX2-induced oxidative burst, which allows histone-modifying enzymes, such as neutrophil elastase (NE) and peptidyl-arginine deaminase 4 (PAD4), to enter the nucleus. Stage 2 involves the entropic swelling of chromatin, rupture of the cell membrane, and seeding of NETs composed of chromatin and granule proteins.

[0006] Inhibitors of NETosis may control chronic neutrophil-driven diseases. A recent phenotypic screen of a large chemical library in human neutrophils identified the compound LDC7559 as an inhibitor of NOX2-dependent NETosis (Sollberger, G., et al., Sci. Immunol., 2018, 3, eaar6689). It was proposed to target the pore-forming domain of gasdermin D (GSDMD), a protein that mediates a lytic form of cell death called pyroptosis. In macrophages, cleavage of GSDMD by human caspases 1, 4, or 5 (mouse caspases 1 or 11) releases an N-terminal fragment that forms pores in the membrane (Kayagaki, N. and Dixit, V.M., Science, 2010, 366, 688-689). In neutrophils, caspase-4 cleaves GSDMD and extrudes NETs in response to cytosolic lipopolysaccharide (LPS) (Chen, KW, et al., Sci. Immunol., 2018, 3, aar6676). GSDMD can also be cleaved by neutrophil-specific proteases such as NE and cathepsin G (Burgener, SS, et al., Cell Rep., 2019, 27, 3646-3656; and Kambara, H., et al., Cell Rep., 2018, 22, 2924-2936). It remained unclear how LDC7559 inhibited GSDMD to prevent NETosis and why LDC7559 inhibited NOX2-dependent but not NOX2-independent NETosis.

[0007] Currently, there is a need for compounds and methods that are useful for agonizing the glycolytic enzyme phosphofructokinase-1 hepatic type (PFKL).There is also a need for compounds that are useful for suppressing NOX2-dependent oxidative burst.Such compounds and methods will be useful for treating diseases including diabetes and cancer. Summary of the Invention

[0008] overview LDC7559 and the compounds of formula (I) were found to agonize PFKL and inhibit the NOX2-dependent oxidative burst.

[0009] In a first embodiment (embodiment 1; abbreviated as E1), the present invention provides a compound of formula (I): TIFF2024527604000002.tif18170 (in the formula, R 1 -NR a R b or one or more groups R c is a 5-10 membered heteroaryl optionally substituted with R 2 is one or more groups R r or R 2 is one or more groups R s or R 2 is one or more groups R z is a 3- to 10-membered heterocycle optionally substituted with R a is (C1-C6)alkyl, (C3-C6)cycloalkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1-C6)alkanoyl, (C3-C6)cycloalkyl(C1-C6)alkyl, (C2-C6)alkynylcarbonyl, a 3- to 6-membered heterocycle, or one or more groups R f and each of (C1-C6)alkyl, (C3-C6)cycloalkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1-C6)alkanoyl, (C3-C6)cycloalkyl(C1-C6)alkyl, (C2-C6)alkynylcarbonyl, and the 3-6 membered heterocycle is selected from the group consisting of halo, hydroxy, cyano, (C2-C6)alkenyl, (C2-C6)alkynyl, C(=O)NR m R n , as well as halo, hydroxy, cyano, -NR m R n , and -C(=O)NR m R n(C1-C6)alkyl, optionally substituted with one or more groups independently selected from the group consisting of: R b is H or (C1-C6)alkyl; Each R c is cyano, -NR d R e , -C(=O)NR d R e , (C1-C6)alkyl, (C3-C6)cycloalkyl, (C3-C6)cycloalkyl(C1-C6)alkyl, (C1-C6)alkoxy, and (C1-C6)alkanoyl, each of which is optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, and cyano; R d and R e are each independently selected from the group consisting of H and (C1-C6)alkyl; or R d and R e together with the nitrogen to which they are attached form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C1-C6)alkyl; Each R f is halo, hydroxy, cyano, -NR g R h , -C(=O)NR g R h , as well as halo, hydroxy, carboxy, -NR g R h , -C(=O)NR g R h (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of cyano; R g and R hare each independently selected from the group consisting of H and (C1-C6)alkyl; or R g and R h together with the nitrogen to which they are attached form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C1-C6)alkyl; R m is H or (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, cyano and oxo; R n is H or (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, cyano and oxo; Each R r is halo, hydroxy, cyano, -NR t R u , -C(=O)NR t R u , -S(O)NR t R u , (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkylthio, -N(H)S(O)R x , -S(O)2R x , (C2-C6)alkenyl, and (C2-C6)alkynyl, each of (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkylthio, (C2-C6)alkenyl, and (C2-C6)alkynyl is independently selected from the group consisting of halo, hydroxy, carboxy, -NR t R u , -C(=O)NR t R u , -S(O)NR t R u , -S(O)2R x and cyano; Each R s -Halo, Cyano, -NR v R w, -C(=O)NR v R w , -S(O)NR v R w , (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkanoyl, (C1-C6)alkylthio, 3- to 6-membered heterocycle, and -S(O)R y and each (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkanoyl, 3- to 6-membered heterocycle, and (C1-C6)alkylthio is independently selected from the group consisting of halo, hydroxy, carboxy, -NR t R u , -C(=O)NR t R u , S(O)NR v R w , -S(O)2R y and cyano; R t and R u are each independently selected from the group consisting of H, (C-C) alkyl, (C-C) alkanoyl, and (C-C) alkynylcarbonyl; or R t and R u together with the nitrogen to which they are attached form a 3-6 membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C1-C6)alkyl; R v and R w are each independently selected from the group consisting of H, (C1-C6) alkyl, and (C1-C6) alkanoyl; or R v and R w together with the nitrogen to which they are attached form a 3-6 membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C1-C6)alkyl; R xis H or (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, cyano and oxo; R y is H or (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, cyano and oxo; Each R z -Oxo, halo, hydroxy, and halo, hydroxy, carboxy, -NR t R u , -C(=O)NR t R u , S(O)NR v R w , -S(O)2R y , cyano, and oxo; The present invention provides a compound of the present invention,

[0010] Further embodiments (E2 to E81) of the first embodiment of the present invention will be described below.

[0011] E2.R 1 But -NR a R b or one or more groups R c is a 5-10 membered heteroaryl optionally substituted with R 2 has one or more groups R r or R 2 has one or more groups R s or R 2 has one or more groups R z is a 9-membered heterocycle optionally substituted with R ais (C1-C6) alkyl, (C3-C6) cycloalkyl, (C2-C6) alkynyl, (C1-C6) alkanoyl, (C2-C6) alkynylcarbonyl, a 3- to 6-membered heterocycle, or one or more groups R f and each (C1-C6)alkyl and (C3-C6)cycloalkyl is selected from the group consisting of halo, cyano, (C2-C6)alkynyl, C(=O)NR m R n and (C1-C6)alkyl optionally substituted with one or more hydroxy; R b is H or (C1-C6)alkyl; Each R c But, cyano, -NR d R e and (C1-C6)alkyl optionally substituted with one or more cyano; R d and R e are H, respectively; Each R f is halo, hydroxy, cyano, -C(=O)NR g R h and (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy and carboxy; R g and R h are H, respectively; R m is H; R n is H; Each R r But halo, hydroxy, cyano, -NR t R u , -C(=O)NR t R u , -S(O)NR t R u , (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkylthio, -N(H)S(O)Rx , -S(O)2R x and (C2-C6)alkynyl, each of (C1-C6)alkyl and (C2-C6)alkynyl being independently selected from the group consisting of hydroxy, -NR t R u , -C(=O)NR t R u and cyano; Each R s But halo, cyano, -NR v R w , -C(=O)NR v R w , (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkanoyl, (C1-C6)alkylthio, 3- to 6-membered heterocycle, and -S(O)R y and each (C-C)alkyl is independently selected from the group consisting of halo and -NR t R u and optionally substituted with one or more groups independently selected from the group consisting of: R t and R u are each independently selected from the group consisting of H, (C-C) alkyl, (C-C) alkanoyl, and (C-C) alkynylcarbonyl; or R t and R u together with the nitrogen to which they are attached form a 3-6 membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C1-C6)alkyl; R v and R w are each independently selected from the group consisting of H, (C1-C6)alkyl, and (C1-C6)alkanoyl; R x is (C1-C6) alkyl; R y is (C1-C6) alkyl; Each R zis independently selected from the group consisting of oxo and (C1-C6)alkyl, or a prodrug thereof, or a pharma- ceutically acceptable salt thereof.

[0012] E3.R 1 Ga-NR a R b A compound according to E1 or E2, or a prodrug, or a pharma- ceutically acceptable salt thereof, wherein:

[0013] E4.R a is (C1-C6)alkanoyl optionally substituted with one or more groups independently selected from the group consisting of halo and hydroxy, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0014] E5.R a A compound according to E1, E2 or E3, or a prodrug, or a pharma- ceutically acceptable salt thereof, wherein is (C1-C6)alkanoyl.

[0015] E6.R a A compound according to E1, E2 or E3, or a prodrug, or a pharma- ceutically acceptable salt thereof, wherein is acetyl.

[0016] E7.R a is halo, hydroxy, cyano, (C2-C6)alkenyl, (C2-C6)alkynyl, C(=O)NR m R n , as well as halo, hydroxy, cyano, -NR m R n , and -C(=O)NR m R n or a prodrug or a pharma- ceutically acceptable salt thereof.

[0017] E8.Ra is halo, hydroxy, cyano, (C2-C6)alkenyl, (C2-C6)alkynyl, C(=O)NR m R n , as well as halo, hydroxy, cyano, -NR m R n , and -C(=O)NR m R n and (C1-C6)cycloalkyl optionally substituted with one or more groups independently selected from the group consisting of: (C1-C6)alkyl optionally substituted with one or more groups independently selected from the group consisting of:

[0018] E9.R a Cyano, (C2-C6)alkynyl, C(=O)NR m R n and (C1-C6)alkyl, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0019] E10.R a has one or more groups R f or a prodrug or a pharma- ceutically acceptable salt thereof.

[0020] E11.R a But the following: A compound according to E1, E2 or E3, or a prodrug or a pharma- ceutically acceptable salt thereof, selected from the group consisting of TIFF2024527604000003.tif149170.

[0021] E12.R b is H, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0022] E13.R 1 has one or more groups R c The compound according to E1 or E2, or a prodrug or a pharma- ceutically acceptable salt thereof, wherein R is 5-10 membered heteroaryl optionally substituted with R.

[0023] E14.R 1 has one or more groups R c or a prodrug, or a pharma- ceutically acceptable salt thereof.

[0024] E15.R 1 But, cyano, -NR d R e and (C1-C6)alkyl, each (C1-C6)alkyl being optionally substituted with one or more groups independently selected from the group consisting of halo and cyano, or a prodrug, or a pharma- ceutically acceptable salt thereof.

[0025] E16.R 1 But the following: A compound according to E1 or E2, or a prodrug or a pharma- ceutically acceptable salt thereof, selected from the group consisting of TIFF2024527604000004.tif85170.

[0026] E17.R 2 has one or more groups R s or a prodrug or a pharma- ceutically acceptable salt thereof.

[0027] E18.R 2 has one or more groups R ror a prodrug or a pharma- ceutically acceptable salt thereof.

[0028] E19.Each R r But halo, cyano, -NR t R u , -C(=O)NR t R u , -S(O)NR t R u , (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)alkoxy, (C1-C6)alkylthio, -N(H)S(O)R x , and -S(O)R x wherein each (C-C)alkyl is independently selected from hydroxy and -NR t R u or a prodrug or a pharma- ceutically acceptable salt thereof, optionally substituted with one or more groups independently selected from the group consisting of:

[0029] E20.R 2 is selected from the group consisting of: TIFF2024527604000005.tif191170

[0030] E21.R 2 has one or more groups R s or a prodrug or a pharma- ceutically acceptable salt thereof.

[0031] E22.Each R s But, cyano, -NR v R w , -C(=O)NR v R w , (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) alkoxy, (C1-C6) alkylthio, -S(O)2R y and (C1-C6)alkanoyl, each (C1-C6)alkyl being independently selected from the group consisting of halo and -NR t R u or a prodrug or a pharma- ceutically acceptable salt thereof, optionally substituted with one or more groups independently selected from the group consisting of:

[0032] E23.R 2 But the following: A compound described in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, or E16, or a prodrug or a pharma- ceutically acceptable salt thereof, selected from the group consisting of TIFF2024527604000006.tif188170.

[0033] E24.R 2 has one or more groups R z A compound according to E1, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, or E16, or a prodrug or a pharma- ceutically acceptable salt thereof, wherein R is a 3- to 10-membered heterocycle optionally substituted with R.

[0034] E25.R 2 has one or more groups R z A compound according to E1, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, or E16, or a prodrug or a pharma- ceutically acceptable salt thereof, wherein R is a 5- to 10-membered heterocycle optionally substituted with R.

[0035] E26.Each R z is oxo, or (C1-C3)alkyl, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0036] E27.R 2 But the following: A compound according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, or E16, or a prodrug or a pharma- ceutically acceptable salt thereof, selected from the group consisting of TIFF2024527604000007.tif66170.

[0037] E28.R 2 has one or more groups R r R is a 6- to 12-membered aryl substituted with r is optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, cyano, and oxo; t R u or (C2-C6)alkynyl, R t is H;R u is (C2-C6)alkynyl-C(=O)- optionally substituted with one or more groups independently selected from the group consisting of halo, hydroxy, cyano and oxo, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0038] E29.R 2 has one or more groups R r phenyl substituted with R ris (C2-C6)alkynyl optionally substituted with one or more groups independently selected from the group consisting of hydroxy and cyano, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0039] E30.R 2 but, TIFF2024527604000008.tif13170; R r is (C2-C6)alkynyl substituted with hydroxy, or a prodrug or a pharma- ceutically acceptable salt thereof.

[0040] E31.R 2 But the following: A compound described in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, or E16, or a prodrug or a pharma- ceutically acceptable salt thereof, selected from the group consisting of TIFF2024527604000009.tif22170.

[0041] E32. Below: A compound, prodrug, or pharma- ceutically acceptable salt according to E1, selected from the group consisting of TIFF2024527604000010.tif100170 and prodrugs and pharma- ceutically acceptable salts thereof.

[0042] E33. Below: A compound, prodrug, or pharma- ceutically acceptable salt according to E1, selected from the group consisting of TIFF2024527604000011.tif213170 and prodrugs and pharma- ceutically acceptable salts thereof.

[0043] E34. Below: A compound, prodrug, or pharma- ceutically acceptable salt according to E1, selected from the group consisting of: TIFF2024527604000012.tif239170TIFF2024527604000013.tif207170 and prodrugs and pharma- ceutically acceptable salts thereof.

[0044] E35. Below: TIFF2024527604000014.tif223170TIFF2024527604000015.tif238170TIFF2024527604000016.tif243170TIFF2024527604000017.tif255170TIFF2024527604000018.tif253170TIFF2024527604000019.tif32170 and prodrugs and pharma- ceutically acceptable salts thereof.

[0045] E36. Below: A compound, prodrug, or pharma- ceutically acceptable salt according to E1, selected from the group consisting of TIFF2024527604000020.tif85170 and prodrugs and pharma- ceutically acceptable salts thereof.

[0046] E37. Below: A compound, prodrug, or pharma- ceutically acceptable salt according to E1, selected from the group consisting of TIFF2024527604000021.tif30170 and prodrugs and pharma- ceutically acceptable salts thereof.

[0047] E38. A prodrug according to E1, which is a compound of formula (I) containing a hydroxy group converted into a prodrug group that increases the aqueous solubility of the compound, or a pharma- ceutically acceptable salt thereof.

[0048] E39. Hydroxy group is phosphate A prodrug according to E38, or a pharma- ceutically acceptable salt thereof, which has been converted to a prodrug group selected from the group consisting of: TIFF2024527604000022.tif19170.

[0049] E40. or less: A prodrug as described in E1, selected from the group consisting of TIFF2024527604000023.tif136170 and pharma- ceutically acceptable salts thereof.

[0050] E41. Below: A compound according to E1, E2, E3, E4, E5, E6, E12, E18, or E19, or a prodrug or a pharma- ceutically acceptable salt thereof, which is not a compound of TIFF2024527604000024.tif22170.

[0051] E42.R 1 is not 3-pentynoylamino or amino; 2 A compound according to E1, E2, E3, E12, E18, or E19, or a prodrug or a pharma- ceutically acceptable salt thereof, wherein is 2-methoxyphenyl.

[0052] E43.R 2 A compound according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E18, or E19, or a prodrug or a pharma- ceutically acceptable salt thereof, provided that is not 2-methoxyphenyl.

[0053] E44. Below: TIFF2024527604000025.tif248170TIFF2024527604000026.tif228170TIFF2024527604000027.tif244170TIFF2024527604000028.tif208170TIFF2024527604000029.tif217170TIFF2024527604000030.tif94170, or a prodrug thereof, or a pharma- ceutically acceptable salt thereof.

[0054] E45. A pharmaceutical composition comprising a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, and a pharma- ceutically acceptable excipient.

[0055] E46. A method for treating a disease associated with activity of the glycolytic enzyme, phosphofructokinase-1 hepatic type in an animal, comprising administering to the animal a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44.

[0056] E47. A method for treating cancer in an animal (e.g., a mammal such as a human), comprising administering to the animal a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44.

[0057] E48. A method for treating diabetes in an animal (e.g., a mammal such as a human), comprising administering to the animal a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44.

[0058] E49. A method for treating a caspase-associated autoinflammatory condition in an animal (e.g., a mammal such as a human), comprising administering to the animal a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44.

[0059] E50. The method of E49, wherein the caspase-associated autoinflammatory condition is sepsis or septic shock.

[0060] E51. A method for treating a disease or condition in an animal selected from the group consisting of pulmonary disease, systemic autoimmune disease, atherosclerosis, thrombosis, multiple sclerosis, Alzheimer's disease, psoriasis, and pulmonary fibrosis, comprising administering to an animal a therapeutically effective amount of E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E26, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, E44, E45, E46, E47, E48, E49, E50, E51, E52, E53, E54, E55, E56, E57, E58, E59, E60, E61, E62, E63, E64, E65, E66, E67, E68, E70, E72, E74, E76, E78, E80, E81, E82, E83, E84, E85, E86, E87, E88, E90, E89, E91, E92, E93, E94, E95, E96, E97, E98, E99, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E103, E104, E105, E106, E107, 7, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, comprising administering to an animal a compound, prodrug, or pharma- ceutically acceptable salt thereof.

[0061] E52. The method of E51, wherein the pulmonary disease is acute respiratory distress syndrome ARDS, chronic obstructive pulmonary disease COPD, or bronchiectasis.

[0062] E53. The method of E51, wherein the disease or condition is thrombosis.

[0063] E54. A method for treating a disease or condition associated with activity of a homologous PFK enzyme, comprising administering to an animal a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44.

[0064] E55. The method of E54, wherein the disease or condition is associated with activity of PFKM (muscle type).

[0065] E56. The method of E54, wherein the disease or condition is associated with activity of PFKM (platelet type).

[0066] E57. A compound of formula (I), prodrug, or pharma- ceutically acceptable salt thereof as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for use in medical therapy.

[0067] E58. A compound of formula (I) as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, prodrug, or pharma- ceutically acceptable salt, for the prophylactic or therapeutic treatment of a disease associated with the activity of the glycolytic enzyme, phosphofructokinase-1 hepatic type, in an animal.

[0068] E59. A compound of formula (I) as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, or a prodrug or a pharma- ceutically acceptable salt thereof, for the prophylactic or therapeutic treatment of cancer.

[0069] E60. A compound of formula (I), prodrug, or pharma- ceutically acceptable salt thereof as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for the prophylactic or therapeutic treatment of diabetes mellitus.

[0070] E61. A compound of formula (I) as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, prodrug, or pharma- ceutically acceptable salt, for the prophylactic or therapeutic treatment of a caspase-associated autoinflammatory condition.

[0071] E62. The compound, prodrug, or pharma- ceutically acceptable salt according to E61, wherein the caspase-associated autoinflammatory condition is sepsis or septic shock.

[0072] E63. A compound of formula (I) as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, prodrug, or pharma- ceutically acceptable salt, for the prophylactic or therapeutic treatment of a disease or condition selected from the group consisting of pulmonary disease, systemic autoimmune disease, atherosclerosis, thrombosis, multiple sclerosis, Alzheimer's disease, psoriasis, and pulmonary fibrosis.

[0073] E64. The compound, prodrug, or pharma- ceutically acceptable salt according to E63, wherein the pulmonary disease is acute respiratory distress syndrome ARDS, chronic obstructive pulmonary disease COPD, or bronchiectasis.

[0074] E65. The compound, prodrug, or pharma- ceutically acceptable salt of E63, wherein the pulmonary disease is thrombosis.

[0075] E66. A compound of formula (I) as defined in E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44, prodrug, or pharma- ceutically acceptable salt, for the prophylactic or therapeutic treatment of a disease or condition associated with the activity of a homologous PFK enzyme.

[0076] E67. The compound, prodrug, or pharma- ceutically acceptable salt according to E66, wherein the disease or condition is associated with activity of PFKM (muscle type).

[0077] E68. The compound, prodrug, or pharma- ceutically acceptable salt according to E66, wherein the disease or condition is associated with activity of PFKM (platelet type).

[0078] E69. Use of a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for the preparation of a medicament for the treatment of a disease associated with the activity of the glycolytic enzyme, phosphofructokinase-1 hepatic type, in an animal (e.g., a mammal such as a human).

[0079] E70. Use of a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for the preparation of a medicament for the treatment of cancer in an animal (e.g., a mammal such as a human).

[0080] E71. Use of a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for the preparation of a medicament for the treatment of diabetes in an animal (e.g., a mammal such as a human).

[0081] E72. Use of a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for the preparation of a medicament for the treatment of a caspase-associated autoinflammatory disease in an animal (e.g., a mammal such as a human).

[0082] E73. The use of E72, wherein the caspase-associated autoinflammatory condition is sepsis or septic shock in an animal (e.g., a mammal such as a human).

[0083] E74. E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13 for preparing a medicament for the treatment of a disease or condition selected from the group consisting of pulmonary disease, systemic autoimmune disease, atherosclerosis, thrombosis, multiple sclerosis, Alzheimer's disease, psoriasis, and pulmonary fibrosis in an animal (e.g., a mammal such as a human). , E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44.

[0084] E75. The use of E74, wherein the pulmonary disease is acute respiratory distress syndrome ARDS, chronic obstructive pulmonary disease COPD, or bronchiectasis.

[0085] E76. The use of E74, wherein the disease or condition is thrombosis.

[0086] E77. Use of a compound, prodrug, or pharma- ceutically acceptable salt according to E1, E2, E3, E4, E5, E6, E7, E8, E9, E10, E11, E12, E13, E14, E15, E16, E17, E18, E19, E20, E21, E22, E23, E24, E25, E36, E27, E28, E29, E30, E31, E32, E33, E34, E35, E36, E37, E38, E39, E40, E41, E42, E43, or E44 for the preparation of a medicament for the prophylactic or therapeutic treatment of a disease or condition associated with the activity of a homologous PFK enzyme in an animal (e.g., a mammal such as a human).

[0087] E78. The use of E77, wherein the disease or condition is associated with activity of PFKM (muscle type).

[0088] E79. The use of E77, wherein the disease or condition is associated with activity of PFKP (platelet type).

[0089] E80. The method of E47, the compound, prodrug, or pharma- ceutically acceptable salt of E59, or the use of E70, wherein the cancer is selected from the group consisting of brain cancer, breast cancer, lung cancer, bladder cancer, cervical cancer, skin cancer, oral cancer, pharyngeal cancer, colon cancer, liver cancer, appendicitis cancer, stomach cancer, pancreatic cancer, prostate cancer, esophageal cancer, blood cancer, thyroid cancer, uterine cancer, and head and neck cancer.

[0090] E81. A process or synthetic intermediate disclosed herein useful for preparing a compound of formula (I) or a prodrug thereof, or a pharma- ceutically acceptable salt thereof.

[0091] E82. The invention described herein. [Brief description of the drawings]

[0092] [Figure 1]LDC7559 and the compound of Example 6 (NA-11) inhibit NETosis independent of GSDMD but induce an identical phenotype in neutrophils. LDH (A) or IL-1β (B) released from primary human monocytes. Bars represent the mean ± sem of monocytes from three donors. (C) Western blot of monocytes 1 h after electroporation with LPS. FL, full-length GSDMD. NT, N-terminal fragment of GSDMD. Results representative of three independent experiments. (D) Coomassie blue staining of recombinant GSDMD. Results representative of three independent experiments. (E) TR-FRET assay measuring europium-labeled biotin released from liposomes exposed to caspase-4 and GSDMD. Symbols represent the mean ± sd of three independent experiments. (F-G) Percentage of polymorphonuclear leukocytes (PMNs) undergoing NETosis induced by PMA (F) or the indicated stimuli (G). Data are means ± sd of cells from three donors. (H) ROS production by PMNs. Bars represent means ± sem of PMNs from three donors. If P<0.05, P values ​​(2-way ANOVA, means compared to medium only) are indicated. (I) Percentage of PMNs undergoing PMA-induced NETosis. The x-axis indicates the addition of NA-11 versus the addition of PMA (t=0). Bars represent means ± sem of PMNs from three donors. If P<0.05, P values ​​(1-way ANOVA, means compared to medium only) are indicated. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0093] Detailed Description In neutrophils, NADPH generated through the pentose phosphate pathway fuels the NADPH oxidase NOX2 to generate reactive oxygen species and kill invading pathogens. Excessive NOX2 activity can exacerbate inflammation, as in acute respiratory distress syndrome (ARDS). Utilizing two unbiased chemical proteomics strategies, we showed that the small molecules LDC7559 and Example 6 inhibit the NOX2-dependent oxidative burst in neutrophils by activating the glycolytic enzyme phosphofructokinase-1 liver type (PFKL) and attenuating flux through the pentose phosphate pathway. Thus, neutrophils treated with Example 6 exhibited defects in NOX2-dependent output, including neutrophil cell death (NETosis) and tissue damage. The high-resolution structure of PFKL confirmed the binding of Example 6 to the AMP / ADP allosteric activation site, explaining why Example 6 failed to agonize phosphofructokinase-1 platelet type (PFKP) or muscle type (PFKM). Selective activation of different phosphofructokinase-1 isoforms may provide new therapeutic paradigms for treating diseases such as ARDS, diabetes and cancer.

[0094] The following definitions are used unless otherwise stated: Halo or halogen is fluoro, chloro, bromo or iodo. Alkyl, alkoxy, alkenyl, alkynyl, etc. refer to both straight chain and branched groups. However, references to individual radicals such as propyl include only the straight chain group; branched chain isomers such as isopropyl are specifically referred to.

[0095] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon radical having the specified number of carbon atoms (i.e., C 1-8means 1 to 8 carbons. Examples include (C1-C8) alkyl, (C2-C8) alkyl, C1-C6) alkyl, (C2-C6) alkyl, and (C3-C6) alkyl. Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, iso-butyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and higher homologs and isomers.

[0096] The term "alkenyl" refers to an unsaturated alkyl radical having one or more double bonds. Examples of such unsaturated alkyl groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl) and higher homologs and isomers.

[0097] The term "alkynyl" refers to an unsaturated alkyl radical having one or more triple bonds. Examples of such unsaturated alkyl groups are ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers.

[0098] The term "alkoxy" refers to an alkyl group attached to the remainder of the molecule through an oxygen atom ("oxy").

[0099] The term "alkylthio" refers to an alkyl group attached to the remainder of the molecule via a thio group.

[0100] The term "cycloalkyl" refers to a saturated or partially unsaturated (non-aromatic) all-carbocyclic ring having 3 to 8 carbon atoms (i.e., (C3-C8 carbocyclic rings). The term also includes multiple condensed saturated all-carbocyclic ring systems (e.g., ring systems containing 2, 3, or 4 carbocyclic rings). Thus, carbocyclic rings include bicyclic carbocyclic rings (e.g., bicyclic carbocyclic rings having about 3 to 15 carbon atoms, about 6 to 15 carbon atoms, or 6 to 12 carbon atoms, such as bicyclo[3.1.0]hexane and bicyclo[2.1.1]hexane) and polycyclic carbocyclic rings (e.g., tricyclic and tetracyclic carbocyclic rings having up to about 20 carbon atoms). Rings of multiple condensed ring systems may be condensed, if valence requirements permit. They may be connected to each other through a fused bond, a spiro bond, and a bridged bond. For example, polycyclic carbocycles may be connected to each other through a single carbon atom to form a spiro bond (e.g., spiropentane, spiro[4,5]decane, etc.), through two adjacent carbon atoms to form a fused bond (e.g., decahydronaphthalene, norsabinane, norcarane, etc. carbocycles), or through two non-adjacent carbon atoms to form a bridged bond (e.g., norbornane, bicyclo[2.2.2]octane, etc.). Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptane, pinane, and adamantane.

[0101] The term "aryl" as used herein refers to a single all-carbon aromatic ring or a multiple condensed all-carbon ring system, where at least one of the rings is aromatic. For example, in certain embodiments, an aryl group has 6-20 carbon atoms, 6-14 carbon atoms, 6-12 carbon atoms, or 6-10 carbon atoms. Aryl includes phenyl groups. Aryl also includes multiple condensed ring systems (e.g., ring systems containing 2, 3, or 4 rings) having about 9-20 carbon atoms, where at least one ring is aromatic and the other rings may or may not be aromatic (i.e., cycloalkyl). The rings of a multiple condensed ring system may be bonded to each other through fused bonds, spiro bonds, and bridge bonds, where valence requirements permit. The point of attachment of a multiple condensed ring system may be at any position of the ring system, including the aromatic or carbocyclic portions of the rings, as defined above. Non-limiting examples of aryl groups include, but are not limited to, phenyl, indenyl, indanyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, and the like.

[0102] The term "heterocycle" refers to a single saturated or partially unsaturated ring having at least one atom other than carbon in the ring, the atom being selected from the group consisting of oxygen, nitrogen, and sulfur; the term also includes multiple fused ring systems having at least one such saturated or partially unsaturated ring, which are further described below. Thus, the term includes a single saturated or partially unsaturated ring (e.g., a 3-, 4-, 5-, 6-, or 7-membered ring) of about 1-6 carbon atoms and about 1-3 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur in the ring. The sulfur and nitrogen atoms may be present in their oxidized forms. Exemplary heterocycles include, but are not limited to, azetidinyl, tetrahydrofuranyl, and piperidinyl. The term "heterocycle" also includes multiple fused ring systems (e.g., ring systems containing 2, 3, or 4 rings), and a single heterocycle (as defined above) may be fused with one or more groups selected from cycloalkyl, aryl, and heterocycle to form multiple fused ring systems. The rings of a multiple fused ring system may be bonded to each other through fused bonds, spiro bonds, and bridged bonds, if valence requirements permit. The individual rings of a multiple fused ring system may be bonded to each other in any order. It should also be understood that the point of attachment of a multiple fused ring system (as defined above for heterocycle) may be at any position of the multiple fused ring system, including the heterocyclic, aryl, and carbocyclic portions of the ring. In one embodiment, the term heterocycle includes a 3-15 membered heterocycle. In one embodiment, the term heterocycle includes a 3-10 membered heterocycle. In one embodiment, the term heterocycle includes a 3-8 membered heterocycle. In one embodiment, the term heterocycle includes a 3-7 membered heterocycle. In one embodiment, the term heterocycle includes 3-6 membered heterocycles. In one embodiment, the term heterocycle includes 4-6 membered heterocycles. In one embodiment, the term heterocycle includes 3-10 membered monocyclic or bicyclic heterocycles containing 1-4 heteroatoms. In one embodiment, the term heterocycle includes 3-8 membered monocyclic or bicyclic heterocycles containing 1-3 heteroatoms. In one embodiment, the term heterocycle includes 3-6 membered monocyclic or bicyclic heterocycles containing 1-2 heteroatoms.In one embodiment, the term heterocycle includes 4-6 membered monocyclic or bicyclic heterocycles containing 1-2 heteroatoms. Exemplary heterocycles include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, homopiperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, tetrahydrofuranyl, dihydrooxazolyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,2,3,4-tetrahydroquinolyl, benzoxazinyl, dihydrooxazolyl, chromanyl, 1,2-dihydropyridinyl, 2,3-dihydrobenzofuranyl, These include, but are not limited to, 1,3-benzodioxolyl, 1,4-benzodioxanyl, spiro[cyclopropane-1,1'-isoindolinyl]-3'-one, isoindolinyl-1-one, 2-oxa-6-azaspiro[3.3]heptanyl, imidazolidin-2-one imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, and 1,4-dioxane.

[0103] As used herein, the term "heteroaryl" refers to a single aromatic ring having at least one atom other than carbon in the ring, where the atom is selected from the group consisting of oxygen, nitrogen, and sulfur, and "heteroaryl" also includes multiple condensed ring systems having at least one such aromatic ring, which are further described below. Thus, "heteroaryl" includes a single aromatic ring consisting of about 1-6 carbon atoms and about 1-4 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur. The sulfur and nitrogen atoms may also be present in oxidized form when the ring is aromatic. Exemplary heteroaryl ring systems include, but are not limited to, pyridyl, pyrimidinyl, oxazolyl, or furyl. The term "heterocycle" also includes multiple condensed ring systems (e.g., ring systems containing 2, 3, or 4 rings), and a single heterocycle (as defined above) may be fused with one or more groups selected from cycloalkyl, aryl, and heterocycle to form multiple condensed ring systems. It should be understood that the attachment point of the heteroaryl or heteroaryl multiple condensed ring system can be any suitable atom of the heteroaryl or heteroaryl multiple condensed ring system, including carbon atoms and heteroatoms (e.g., nitrogen). Exemplary heteroaryls include, but are not limited to, pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, furyl, oxadiazolyl, thiadiazolyl, quinolyl, isoquinolyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalyl, and quinazolyl.

[0104] As used herein, the term "alkoxycarbonyl" refers to the group (alkyl)-OC(=O)-, where the term alkyl has the meaning defined herein.

[0105] As used herein, the term "alkanoyloxy" refers to the group (alkyl)-C(=O)-O-, where the term alkyl has the meaning defined herein.

[0106] As used herein, the term "heteroatom" includes oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).

[0107] The term "protecting group" as used herein refers to a substituent that is commonly used to block or protect a particular functional group on a compound. For example, an "amino protecting group" is a substituent added to an amino group that blocks or protects the amino functionality in a compound. Suitable amino protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ), and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a "hydroxy protecting group" refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A "carboxy protecting group" refers to a substituent of a carboxy group that blocks or protects the carboxy functionality. Common carboxy protecting groups include phenylsulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2-(diphenylphosphino)-ethyl, nitroethyl, and the like. For a general description of protecting groups and their uses, see PGM Wuts and TW Greene, Greene's Protective Groups in Organic Synthesis, 4th ed., Wiley-Interscience, New York, 2006.

[0108] As used herein, crossing a bond in a chemical structure TIFF2024527604000031.tif5170 shows the point of attachment of the bond in the chemical structure that is crossed by a wavy bond to the rest of the molecule.

[0109] The term "treat", "treatment" or "treating", insofar as it pertains to a disease or condition, includes inhibiting a disease or condition, eliminating a disease or condition, and / or alleviating one or more symptoms of a disease or condition. The term "treat", "treatment" or "treating" also refers to both therapeutic and / or prophylactic treatments or preventative measures, the purpose being to prevent or slow down (alleviate) the onset or spread of an undesirable physiological change or disorder, such as cancer. For example, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, whether detectable or undetectable, attenuation of the extent of a disease or disorder, a stabilized (i.e., non-worsening) state of a disease or disorder, a delay or slowing of disease progression, an improvement or alleviation of a disease state or disorder, and remission (whether partial or total). "Treat", "treatment" or "treating" can also mean prolonging survival compared to expected survival in the absence of treatment. Those in need of treatment include those already with the disease or disorder, as well as those prone to have the disease or disorder or those having the disease or disorder which is to be prevented. In one embodiment, "treat", "treatment" or "treating" does not include preventing or prophylaxis.

[0110] The phrase "therapeutically effective amount" or "effective amount" includes, but is not limited to, an amount of a compound that (i) treats or prevents a particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein.

[0111] As used herein, the term "mammal" refers to humans, higher non-human primates, rodents, livestock, cattle, horses, pigs, sheep, dogs and cats. In one embodiment, a mammal is a human. As used herein, the term "patient" refers to any animal, including mammals. In one embodiment, the patient is a mammalian patient. In one embodiment, the patient is a human patient.

[0112] The compounds described herein may also exist as tautomers in certain cases. Although only one delocalized resonance structure may be exhibited, all such forms are contemplated within the scope of the present invention.

[0113] The present invention also includes, but is not limited to, deuterium ( 2 It will be understood by one of skill in the art to include any claimed compound that may be enriched at any or all atoms beyond the naturally occurring isotopic ratio with one or more isotopes, such as H or D. As a non-limiting example, a -CH3 group may be replaced with -CD3.

[0114] The pharmaceutical composition of the present invention may include one or more additives.When used in conjunction with the pharmaceutical composition of the present invention, the term "additive" generally refers to additional components that are combined with the compound of formula (I) or its pharma- ceutically acceptable salt to provide the corresponding composition.For example, when used in conjunction with the pharmaceutical composition of the present invention, the term "additive" includes, but is not limited to, the following: carriers, binders, disintegrants, lubricants, sweeteners, flavorings, coatings, preservatives, and dyes.

[0115] Stereochemical definitions and conventions used herein generally follow those of S.P. Parker, ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric or chiral centers and therefore may exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, and atropisomers, and mixtures thereof, such as racemic mixtures, are intended to form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D and L, or R and S, are used to denote the absolute configuration of the molecule around its chiral center or centers. The prefixes d and l or (+) and (-) are used to indicate the sign of rotation of plane polarized light by a compound, with (-) or 1 meaning that the compound is levorotatory. Compounds with the prefix (+) or d are dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A particular stereoisomer is also referred to as an enantiomer, and a mixture of such isomers may be called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or racemate, which may occur when there is no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two optically isomeric species that is devoid of optical activity.

[0116] Those skilled in the art will understand that the compounds of the present invention that have chiral centers can exist and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. It is understood that the present invention encompasses any racemic, optically active, polymorphic or stereoisomer of the compounds of the present invention that have the useful properties described herein, or mixtures thereof, and that methods for preparing optically active forms (e.g., by resolving racemic forms by recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using chiral stationary phases) are well known in the art.

[0117] When a bond in a compound formula herein is drawn in a non-stereochemical manner (e.g., flat), the atom to which the bond is attached includes all stereochemical possibilities. When a bond in a compound formula herein is drawn in a defined stereochemical manner (e.g., bold, bold wedge, dashed line, or dashed wedge), the atom to which the stereochemical bond is attached should be understood to be enriched in the absolute stereoisomer shown, unless otherwise stated. In one embodiment, the compound can be at least 51% of the absolute stereoisomer depicted. In another embodiment, the compound can be at least 60% of the absolute stereoisomer depicted. In another embodiment, the compound can be at least 80% of the absolute stereoisomer depicted. In another embodiment, the compound can be at least 90% of the absolute stereoisomer depicted. In another embodiment, the compound can be at least 95% of the absolute stereoisomer depicted. In another embodiment, the compound can be at least 99% of the absolute stereoisomer depicted.

[0118] Prodrug In addition to salt forms, the present invention provides compounds in prodrug form. As used herein, the term "prodrug" refers to a compound that undergoes easy chemical changes under physiological conditions to provide a compound of formula (I). Furthermore, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to compounds of formula (I) when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0119] Prodrugs of the present invention include compounds in which the free carboxyl group of the compound of the present invention can be derivatized as an amide or alkyl ester. As another example, compounds of the present invention that contain a free hydroxy group can be derivatized as a prodrug by converting the hydroxy group to a group such as, but not limited to, a phosphate ester, hemisuccinate, dimethylaminoacetate, or phosphoryloxymethyloxycarbonyl group, as reviewed in Fleisher, D. et al. (1996) Improved oral drug delivery: solubility limitations overcome by the use of prodrugs Advanced Drug Delivery Reviews, 19:115. Carbamate prodrugs of hydroxy and amino groups are also included, as are carbonate prodrugs, sulfonate esters, and sulfate esters of hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers, where the acyl group can be an alkyl ester that may be substituted with groups including, but not limited to, ether, amine, and carboxylic acid functional groups, or where the acyl group is an amino acid ester as described above, are also included. This type of prodrug is described in J. Med. Chem., (1996), 39:10. More specific examples include prodrugs in which the hydrogen atom of the alcohol group is replaced with (C 1-6 ) alkanoyloxymethyl, 1-((C 1-6 )alkanoyloxy)ethyl, 1-methyl-1-((C 1-6)alkanoyloxy)ethyl, (C 1-6 ) alkoxycarbonyloxymethyl, N-(C 1-6 ) alkoxycarbonylaminomethyl, succinoyl, (C 1-6 ) Alkanoyl, alpha-amino (C 1-4 )alkanoyl, arylacyl, and alpha-aminoacyl or alpha-aminoacyl-alpha-aminoacyl groups, each of which is an L-amino acid that occurs naturally, P(O)(OH), -P(O)(O(C 1-6 ) alkyl) 2 or glycosyl (the radical obtained from removal of the hydroxyl group of the hemiacetate form of a carbohydrate).

[0120] For further examples of prodrug derivatives, see, for example, a) Design of Prodrugs, edited by H. Bundgaard (Elsevier, 1985), and Methods in Enzymology, Vol. 42, p. 309-396, K. Widder, et al. (Academic Press, 1985); b) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 "Design and Application of Prodrugs," edited by H. Bundgaard, p. 113-191 (1991); c) H. Bundgaard, Advanced Drug Delivery Reviews, 8: 1-38 (1992); d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77: 285 (1988); and e) N. Kakeya, et al., Chem. Pharm. Bull., 32: 692 (1984).

[0121] In one embodiment, a compound of formula (I) containing a hydroxy group (e.g., R rProdrugs of compounds of formula (I) containing a hydroxy group may be prepared by converting the hydroxy group to a prodrug group that increases the aqueous solubility of the compound. For example, the hydroxy group may be converted to a phosphate (-OP(=O)(OH)2) or a pharma- ceutically acceptable salt thereof, or the hydroxy group may be converted to the following: TIFF2024527604000032.tif19170.

[0122] In another embodiment, prodrugs of compounds of formula (I), and pharma- ceutically acceptable salts thereof, include R b phosphate (-OP(=O)(OH)2) Such prodrugs can be prepared by converting a (C1) alkyl group substituted with a group selected from the group consisting of: TIFF2024527604000033.tif19170. Such prodrugs undergo a chemical change under physiological conditions to form R b Corresponding compounds of formula (I) are H.

[0123] Non-limiting examples of prodrugs of compounds of formula (I) include the following compounds: TIFF2024527604000034.tif136170 and pharma- ceutically acceptable salts thereof, such as Na, K and Ca salts thereof. Prodrugs of the compounds of formula (I) and pharma- ceutically acceptable salts thereof may be prepared from the corresponding compounds of formula (I) using standard reagents and techniques.

[0124] The specific values ​​listed below for radicals, substituents and ranges are for illustrative purposes only and they do not exclude other defined values ​​or other values ​​within the defined ranges for radicals and substituents. It is understood that two or more values ​​may be combined. It is also understood that the values ​​listed below (or a subset thereof) can be excluded.

[0125] Specifically, (C1-C6) alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl or hexyl; (C3-C6) cycloalkyl can be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; (C3-C6) cycloalkyl(C1-C6) alkyl can be cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl, 2-cyclobutylethyl, 2-cyclopentyl ... (C1-C6)alkoxy can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy or hexyloxy; (C2-C6)alkenyl can be vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1,-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, or or 5-hexenyl; (C2-C6)alkynyl can be ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or 5-hexynyl; (C1-C6)alkanoyl can be acetyl, propanoyl, or butanoyl; (C1-C6)alkoxycarbonyl can be methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, (C1-C6) alkylthio can be methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, pentylthio or hexylthio; (C2-C6) alkanoyloxy can be acetoxy, propanoyloxy, butanoyloxy, isobutanoyloxy, pentanoyloxy or hexanoyloxy; aryl can be phenyl, indenyl or naphthyl;Heteroaryl can be furyl, imidazolyl, triazolyl, triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiazolyl, pyrazolyl, pyrrolyl, pyrazinyl, tetrazolyl, pyridyl (or its N-oxide), thienyl, pyrimidinyl (or its N-oxide), indolyl, isoquinolyl (or its N-oxide) or quinolyl (or its N-oxide);

[0126] Methods for preparing compounds of formula (I) are provided as further embodiments of the present invention. For example, compounds of formula (I) can be prepared using the reaction shown in Scheme 1, where the general radicals have any of the values ​​described herein.

[0127] Scheme 1 TIFF2024527604000035.tif26170

[0128] Bromo compound 100 can be synthesized by the reaction of R 2 For example, a bromo compound can be coupled with compound R in a suitable polar solvent (e.g., DME or dioxane) in the presence of water using a suitable catalyst (e.g., Pd(dppf)Cl). 2 It can be coupled with B(OH)2.

[0129] Compounds of formula (I) may also be prepared using the reaction depicted in Scheme 2.

[0130] Scheme 2 TIFF2024527604000036.tif24170

[0131] Bromo compound 101 can be synthesized by the reaction of R 1 For example, a bromo compound can be coupled with compound R in a suitable polar solvent (e.g., DME or dioxane) in the presence of water using a suitable catalyst (e.g., Pd(dppf)Cl). 1 It can be coupled with B(OH)2.

[0132] The synthetic intermediates described herein that are useful for preparing compounds of formula (I) are provided as further embodiments of the present invention. For example, R 1 and R 2 The intermediate bromo compounds 100 and 101, having any of the values ​​described herein, are synthetic intermediates that can be used to prepare compounds of formula (I). Other synthetic intermediates useful for preparing compounds of formula (I) are described in the Examples.

[0133] When the compound is sufficiently basic or acidic, the salt of the compound of formula (I) may be useful as an intermediate for isolating or purifying the compound of formula (I).In addition, it may be appropriate to administer the compound of formula (I) as a salt of a pharmaceutically acceptable acid or base.Examples of pharmaceutically acceptable salts are organic acid addition salts formed with an acid that forms a physiologically acceptable anion, such as tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, α-ketoglutarate and α-glycerophosphate.Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate and carbonate.

[0134] Salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound, such as an amine, with a suitable acid to give a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium or lithium) or alkaline earth metal (e.g., calcium) salts of carboxylic acids can also be made.

[0135] The compounds of formula (I) may be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient, in a variety of forms adapted to the chosen route of administration, i.e. orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.

[0136] Thus, the compounds may be administered systemically, e.g., orally, in combination with a pharma- ceutically acceptable vehicle, such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the food of the patient's diet. For oral therapeutic administration, the active compounds may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be from about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

[0137] Tablets, troches, pills, capsules, etc. may also contain: Binders such as gum tragacanth, gum acacia, corn starch, or gelatin; Additives such as dicalcium phosphate; Disintegrating agents such as corn starch, potato starch, alginic acid; Lubricants such as magnesium stearate; Sweeteners such as sucrose, fructose, lactose, aspartame, and flavorings such as peppermint, wintergreen oil, cherry flavoring, etc. If the unit dosage form is a capsule, in addition to the above types of materials, it may contain a liquid carrier such as vegetable oil or polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For example, tablets, pills, or capsules may be coated with gelatin, wax, shellac, sugar, and the like. Syrup or elixir may contain the active compound, sucrose or fructose as a sweetener, methyl and propylparabens as preservatives, dyes and flavorings, such as cherry or orange flavoring. Of course, any material used in preparing any unit dosage form should be pharma- ceutically acceptable and substantially non-toxic in the amounts employed.Furthermore, the active compounds may be incorporated into sustained-release preparations and devices.

[0138] The active compound can also be administered intravenously or intraperitoneally by infusion or injection. For intravenous injection, it may be beneficial to convert the compound of formula (I) into a prodrug with increased water solubility. A solution of the active compound or its salt can be prepared in water, optionally mixed with a non-toxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, triacetin, and mixtures thereof and in oils. Under normal conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0139] Pharmaceutical dosage forms suitable for injection or infusion may include sterile aqueous solutions or dispersions or sterile powders containing the active ingredient, which are suitable for the extemporaneous preparation of sterile injectable or injectable solutions or dispersions, optionally encapsulated in liposomes. In all cases, the final dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. Liquid carriers or vehicles may be solvents or liquid dispersion media, including, for example, water, ethanol, polyols (for example, glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof. Proper fluidity may be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. Prevention of microbial activity may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it is preferable to include isotonic agents, for example, sugars, buffers, or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0140] Sterile injectable solutions are prepared by incorporating the required amount of active compound in a suitable solvent with various other ingredients as listed above as required, followed by filter sterilization. In the case of sterile powders for preparing sterile injectable solutions, the preferred preparation method is vacuum drying and freeze-drying technology, which results in a powder of active ingredient and any additional desired ingredients present in the previously sterile-filtered solution.

[0141] In one embodiment, there is provided a formulation comprising a compound of formula (I) or a prodrug thereof, or a salt of a compound of formula (I) or a salt of a prodrug thereof, suitable for intravenous administration.

[0142] For topical administration, the compounds may be applied in pure form, i.e., when they are liquids. However, it is generally desirable to administer them to the skin as a composition or formulation in combination with a dermatologically acceptable carrier, which may be solid or liquid.

[0143] Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina, etc. Useful liquid carriers include water, alcohol or glycol or water-alcohol / glycol blends, in which the compound can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antibacterial agents can be added to optimize the properties for a given application. The resulting liquid composition can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers.

[0144] Thickening agents such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses, or modified mineral materials can also be used with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like for application directly to the user's skin.

[0145] Examples of useful dermatological compositions that can be used to deliver the compounds of formula (I) to the skin are known in the art; see, for example, Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157), and Wortzman (U.S. Pat. No. 4,820,508).

[0146] The useful dosage of the compound of formula (I) can be determined by comparing their in vitro activity and in vivo activity in animal models.The method for extrapolating the effective dosage in mice and other animals to humans is known in the art.See, for example, U.S. Patent No. 4,938,949.

[0147] The amount of the compound or its active salt or derivative required for use in treatment will vary depending not only on the particular salt selected, but also on the route of administration, the nature of the condition being treated, and the age and condition of the patient, and is ultimately at the discretion of the attending physician or clinician.

[0148] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, e.g., two, three, four or more sub-doses per day. The sub-dose itself may, for example, be further divided into several discrete loosely spaced administrations, e.g., by multiple inhalations from an insufflator or by application of multiple drops into the eye.

[0149] The ability of the compound of the present invention to agonize the glycolytic enzyme phosphofructokinase-1 hepatic type can be determined using pharmacological models well known in the art or using the assay described in the following examples.The ability of the compound of the present invention to treat cancer, diabetes, caspase-associated autoinflammatory conditions (e.g., sepsis or septic shock), pulmonary disease (e.g., acute respiratory distress syndrome ARDS, chronic obstructive pulmonary disease COPD, or bronchiectasis), systemic autoimmune disease, atherosclerosis, thrombosis, multiple sclerosis, Alzheimer's disease, psoriasis, pulmonary fibrosis, or diseases or conditions related to the activity of homologous PFK enzyme can also be determined using pharmacological models well known in the art.

[0150] The invention will now be illustrated by the following non-limiting examples. EXAMPLES

[0151] List of Abbreviations AcOH - Acetic acid ACN - Acetonitrile BuLi - Butyl lithium DCM - Dichloromethane DIAD-Diisopropyl azidocarboxylate DME - Dimethoxyethane EDCI-1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide EtOAc - Ethyl acetate EtOH - Ethanol HATU-Hexafluorophosphate azabenzotriazole tetramethyluronium DIPEA-N,N-Diisopropylethylamine HMPA - Hexamethylphosphoric acid triamide MeOH - Methanol MTBE-Methyl tert-butyl ether NIS-N-iodosuccinimide RT-room temperature TBSCl-tert-butyldimethylsilyl chloride TBAF: Tetra-n-butylammonium fluoride TEA - Triethylamine THF - Tetrahydrofuran TLC - Thin Layer Chromatography TosCl-4-Toluenesulfonyl chloride

[0152] Example 1. N-(2-phenyl-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000037.tif34170

[0153] Compound NA-8 (200 mg, 621 μmol), compound 5A (98.4 mg, 807 μmol), K2CO3 (172 mg, 1.24 mmol) and Pd(PPh3)4 (143 mg, 124 μmol) were suspended in DME (4.00 mL) and H2O (2.00 mL). The mixture was heated in a microwave at 120 °C for 1 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R of NA-8) f = 0.35, product R f =0.20) indicated that compound NA-8 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (10.0 mL) and extracted with EtOAc (10.0 mL × 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1, petroleum ether / ethyl acetate = 1 / 1, R of product 1 was obtained. f =0.2). The combined organic layers were concentrated under reduced pressure to give the title compound (75.0 mg, 234 μmol, yield 37.6%, purity 99.5%) as a yellow solid. 1 H NMR:DMSO 400MHz,δ:9.95(s,1H),7.77(d,J=7.2 Hz,2H),7.39(m,J=7.6 Hz,2H),7.26-7.32(m,3H)7.14(d,J=8.4 Hz,1H),6.78(s,1H),5.58(s,2H),5.34(s,2H),2.02(s,3H).LCMS:(M+H +):320.05, calculated value 319.13.

[0154] The intermediate compound NA-8 was prepared as follows. TIFF2024527604000038.tif110170

[0155] a. 1-(Bromomethyl)-2-fluoro-4-nitrobenzene (2)

[0156] To a solution of 1 (120 g, 774 mmol) in CCl4 (840 mL) was added BPO (12.0 g, 49.5 mmol) and NBS (151 g, 851 mmol) at 80° C. The mixture was stirred at 80° C. for 12 h. TLC (petroleum ether / ethyl acetate=20 / 1, R of reaction 1 f = 0.40, product R f =0.30) indicated the reaction was complete. The reaction mixture was cooled to room temperature. The residue was poured into ice water (600 mL) and stirred for 10 min. The aqueous phase was extracted with DCM (300 mL x 3). The combined organic phase was washed with brine (300 mL x 1), dried over anhydrous Na2SO4, filtered and concentrated in vacuum. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 400 / 1 to 30 / 1), then filtered and concentrated in vacuum. Compound 2 (120 g, 513 mmol, 66.3% yield) was obtained as a yellow solid.

[0157] b. Methyl 3-bromo-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazole-5-carboxylate (3)

[0158] To a solution of compound 2 (30.0 g, 146 mmol) in ACN (210 mL) was added CsF (44.5 g, 293 mmol) and compound 2A (37.7 g, 161 mmol). The mixture was stirred at 25° C. for 5 h. TLC (petroleum ether / ethyl acetate=3 / 1, R f1 = 0.30, R of reactants f2 = 0.60, product R f=0.45) indicated that the reaction was incomplete. The mixture was concentrated under reduced pressure to give a crude residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 80 / 1 to 3 / 1). Compound 3 (31.0 g, 86.6 mmol, 59.1% yield) was obtained as a white solid.

[0159] c. (3-bromo-1-(2-fluoro-4-nitrobenzyl)-1H-pyrazol-5-yl)methanol (4)

[0160] To a mixture of compound 3 (62.0 g, 173 mmol) in THF (620 mL) was added DIBAL-H (1.00 M, 346 mL) at 0° C. The mixture was stirred at 15° C. for 1 h under N2 atmosphere. TLC (Plate 1: Petroleum ether / Ethyl acetate=2 / 1, R of reactant 1 f = 0.50, product R f =0) indicated that compound 3 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding saturated NH4Cl solution (300 mL). The resulting mixture was dissolved in EtOAc / EtOH = 1000 mL / 200 mL and filtered to remove insoluble materials. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography ((SiO2, petroleum ether / ethyl acetate = 5 / 1 to 0 / 1, plate 2: petroleum ether / ethyl acetate = 1 / 1, R of compound 4 f =0.35). The combined organic layers were concentrated under reduced pressure to give compound 4 (50.9 g, 154.2 mmol, 89.0% yield) as a white solid.

[0161] d. (1-(4-amino-2-fluorobenzyl)-3-bromo-1H-pyrazol-5-yl)methanol (5)

[0162] To a solution of compound 4 (50.9 g, 154.2 mmol) in MeOH (510 mL) and HO (51.0 mL) was added Fe (34.4 g, 617 mmol) and NH4Cl (41.2 g, 771 mmol). The mixture was stirred at 50° C. for 3 h, then additional Fe (8.61 g, 154 mmol) and NH4Cl (16.5 g, 308 mmol) were added. The mixture was stirred at 50° C. for 22 h. TLC (petroleum ether / ethyl acetate=1 / 1, R of reaction 1 f = 0.57, product R f =0.47), indicating that compound 4 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding saturated NH4Cl solution (200 mL) and then washed with EtOAc (200 mL x 3). The combined organic layers were concentrated under reduced pressure, and the residue was extracted with DCM (200 mL x 3). The combined organic layers were washed with brine (200 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Compound 5 (40.3 g, 134 mmol, 87.0% yield) was obtained as a yellow solid.

[0163] e. 2-Bromo-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-amine (6)

[0164] Compound 5 (23.0 g, 76.6 mmol) was dissolved in HMPA (230 mL) and added to a suspension of NaH (6.13 g, 153 mmol, 60.0% purity) in toluene (2300 mL) under N2. The mixture was stirred at 95 °C under N2 for 3 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R of reactant 1) f = 0.40, product R f=0.56), which indicated that compound 5 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (1000 mL) and extracted with toluene (500 mL × 3). The combined organic layers were washed with brine (500 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound 6 (7.50 g, 26.7 mmol, 34.9% yield) was obtained as a yellow solid.

[0165] fN-(2-bromo-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide (NA-8)

[0166] Compound 6 (4.00 g, 14.3 mmol) was dissolved in THF (120 mL). EtN (2.89 g, 28.6 mmol, 3.98 mL) was added, followed by acetyl chloride (1.35 g, 17.14 mmol, 1.22 mL), and the mixture was stirred at 25° C. for 25 min. TLC (petroleum ether / ethyl acetate=1 / 1, R of compound 6) f = 0.50, R of product 1 f =0.30) indicated that compound 6 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (100 mL) and extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (100 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound NA-8 (3.50 g, 10.8 mmol, yield 75.6%, purity 99.4%) was obtained as a yellow solid. 1H NMR:DMSO 400MHz,δ:9.96(s,1H),7.30(d,J=2 Hz,1H),7.25(d,J=8.4 Hz,1H),7.14(d,J=8.4Hz,1H),6.45(s,1H),5.49(s,2H),5.28(s,2H),2.52-2.55(m,1H),2.02(s,3H).LCMS:(M+H + ):321.9, calculated value 321.01.

[0167] Example 2. N-(2-(3-methoxyphenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000039.tif37170

[0168] NA-8 (200 mg, 621 μmol), compound 6A (189 mg, 1.24 mmol), Na2CO3 (132 mg, 1.24 mmol) and Pd(dppf)Cl2 (45.4 mg, 62.1 μmol) were suspended in H2O (2.00 mL) and dioxane (10.0 mL). The mixture was heated at 95 °C under N2 atmosphere for 16 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R of NA-8) f = 0.30, product R f =0.18) indicated that NA-8 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (10.0 mL) and extracted with EtOAc (10.0 mL × 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound NA-6 (61.0 mg, 169 μmol, yield 27.2%, purity 96.7%) was obtained as a red-brown solid. 1H NMR:DMSO 400MHz,δ:9.96(s,1H),7.24-7.35(m,5H),7.13(d,J=7.8 Hz,1H),6.86(d,J=7.4 Hz,1H),6.79(s,1H),5.57(s,2 H),5.34(s,2H),3.78(s,3H),2.01(s,3H).LCMS:(M+H + ):350.05, calculated value 349.14.

[0169] Example 3. N-(2-(4-methoxyphenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000040.tif34170

[0170] NA-8 (200 mg, 621 μmol), compound 7A (189 mg, 1.24 mmol), Pd(dppf)Cl2 (45.4 mg, 62.1 μmol) and Na2CO3 (132 mg, 1.24 mmol) were suspended in HO (2.00 mL) and dioxane (10.0 mL). The mixture was heated at 95 °C under N2 atmosphere for 16 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R of NA-8) f = 0.30, product R f =0.20) indicated that NA-8 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (10.0 mL) and extracted with EtOAc (10.0 mL × 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound NA-7 (105 mg, 295 μmol, yield 47.5%, purity 98.2%) was obtained as a white solid. 1H NMR:DMSO 400MHz,δ:9.96(s,1H),7.69(d,J=7.8 Hz,2H),7.25-7.29(m,2H),7.13(d,J=8 Hz,1H),6.96(s,1H),6.94(s,1H),6.69(s,1H),5.55(s,2H),5.32(s,2H),3.77(s,3H),3.17-3.31(m,3H),2.01(s,3H).LCMS:(M+H + ):350.05, calculated value 349.14.

[0171] Example 4. N-(3-(7-acetamido-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-2-yl)phenyl)pent-3-ynamide TIFF2024527604000041.tif38170

[0172] To a solution of compound 20 (117 mg, 1.20 mmol) and compound 21 (200 mg, 598 μmol, see Example 7) in DMF (10.0 mL) was added EDCl (229 mg, 1.20 mmol) at 0° C. The mixture was stirred at 18° C. for 2 h. TLC (petroleum ether / ethyl acetate=0 / 1, R f = 0.30, product R f =0.43) and LCMS analysis showed that 21 was completely consumed and the main product peak was detected. The reaction mixture was quenched by adding HO (10.0 mL) and extracted with EtOAc (10.0 mL × 3). The combined organic layers were filtered and concentrated under reduced pressure. The residue was purified by HPLC using a 5 micron Boston Prime C18 column with an internal diameter of 150 mm × 30 mm. The mobile phase consisted of water (containing 0.05% HCl) and a gradient of ACN from 25% to 50% over 10 min. The combined product fractions were concentrated by lyophilization. The title compound (148 mg, 353 μmol, yield 59.0%, purity 98.4%) was obtained as a white solid. 1H NMR:DMSO-d6 400MHz,δ:10.06(s,1H),9.98(s,1H),8.05(s,1H),7.49(d,J=7.45 Hz,1H),7.44(d,J=7.9 Hz,1H),7.25-7.34(m,3H),7.14(m,J=8.10 Hz,1H),6.71(s,1H),5.57(s,2H),5.35(s,2H),4.19(s,7H),3.25-3.29(m,2H),2.52-2.54(m,1H),2.02(s,3H),1.80(m,J=2.64 Hz,3H).LCMS:(M+H + ):415.16, calculated value 414.17.

[0173] Intermediate 20 was prepared as follows.

[0174] a. Pent-3-ynoic acid (20) TIFF2024527604000042.tif17170

[0175] To a solution of compound 19 (3.80 g, 45.2 mmol) in acetone (38.0 mL) was added CrO3 (9.03 g, 90.4 mmol), H2SO4 (31.0 g, 316 mmol) and H2O (22.8 mL). The mixture was stirred at 17 °C for 2 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R of compound 19 was f = 0.46, product R f =0.28), indicating that compound 19 was completely consumed. The reaction was quenched by adding 8.00 mL of isopropanol at 15° C. Then the suspension was filtered through a Celite pad, and the filter cake was washed with EtOAc (8.00 mL×2). The reaction mixture was poured into ice water (8.00 mL) and extracted with ethyl acetate (9.00 mL×3). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. Compound 20 (3.05 g, 31.1 mmol, 68.8% yield) was obtained as a yellow solid.

[0176] Example 5. N-(2-(3-(5-hydroxypent-1-yn-1-yl)phenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000043.tif23170

[0177] Compound 15 (200 mg, 699 μmol), NA-8 (113 mg, 349 μmol), Pd(dppf)Cl2 (25.6 mg, 34.9 μmol) and Na2CO3 (74.0 mg, 699 μmol) were suspended in H2O (1.00 mL) and dioxane (5.00 mL) and heated at 95 °C for 16 h under N2 atmosphere. TLC (petroleum ether / ethyl acetate = 1 / 1, R of NA-8) f = 0.30, product R f =0.20) and LCMS analysis showed that reactant 15 was completely consumed with the formation of one major spot. The reaction mixture was quenched by adding H2O (20.0 mL) and extracted with EtOAc (20.0 mL x 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by HPLC using a 10 micron Kromasil C18 column 100 mm x 40 mm. The mobile phase consisted of water (0.1% TFA) and ACN, a gradient of ACN from 32% to 54% over 10 min. The collected fractions were concentrated by lyophilization. The product was dissolved in aqueous HCl (0.5%, 10.0 mL) and MeCN (2.00 mL), then the liquid was concentrated again by lyophilization. The title compound (67.8 mg, 158 μmol, 45.2% yield, 93.7% purity) was obtained as a white solid. 1 H NMR:CDCl3400MHz,δ:7.84(s,1H),7.77(d,J=6.4 Hz,1H),7.48(s,1H),7.31-7.37(m,2H),7.21-7.25(m,1H),7.18(s,1H),3.86(m,J=6.2 Hz,2H),2.57(m,J=7.1 Hz,2H),2.19(s,3H),1.85-1.93(m,3H),1.27(s,3H).LCMS:(M+H +):402.15, calculated value 401.17.

[0178] Intermediate compound 15 was prepared as follows.

[0179] a. 5-(3-bromophenyl)pent-4-yn-1-ol (14) TIFF2024527604000044.tif29170

[0180] A mixture of compound 13 (2.50 g, 8.84 mmol, 1.13 mL), CuI (101 mg, 530 μmol), Pd(PPh3)2Cl2 (186 mg, 265 μmol) in Et3N (20.0 mL) was stirred at 25 °C under N2 for 30 min. Compound 14A (892 mg, 10.6 mmol) was added and the mixture was stirred at 60 °C under Ar atmosphere for 2 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R of compound 13) was confirmed. f = 0.70, product R f =0.20) indicated that compound 13 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (20.0 mL) and extracted with EtOAc (20.0 mL x 3). The combined organic layers were washed with brine (20.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound 14 (2.00 g, 8.36 mmol, 94.6% yield) was obtained as a brown solid.

[0181] b. 5-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pent-4-yn-1-ol (15) TIFF2024527604000045.tif35170

[0182] A mixture of compound 14 (1.00 g, 4.18 mmol), KOAc (821 mg, 8.36 mmol) and compound 15A (1.27 g, 5.02 mmol) in DMSO (10.0 mL) was degassed and purged with N2 three times. Pd(dppf)Cl2 (306 mg, 418 μmol) was then added and the mixture was stirred at 120 °C under N2 atmosphere for 18 h. TLC (petroleum ether / ethyl acetate = 2 / 1, R of compound 14 was f = 0.35, product R f =0.3) and HPLC analysis showed that reactant 14 was completely consumed with the formation of one major spot. The reaction mixture was quenched by adding H2O (50.0 mL) and extracted with EtOAc (50.0 mL x 3). The combined organic layers were washed with brine (30 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 8 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound 15 (800 mg, 2.80 mmol, 66.8% yield) was obtained as a yellow oil.

[0183] Example 6. N-(2-(2-(5-hydroxypent-1-yn-1-yl)phenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000046.tif33170

[0184] Compound NA-8 (113 mg, 349 μmol), compound 18 (200 mg, 699 μmol), Pd(dppf)Cl2 (25.6 mg, 34.9 μmol) and Na2CO3 (74.1 mg, 699 μmol) were suspended in HO (1.00 mL) and dioxane (5.00 mL) and heated at 95 °C for 16 h under N2 atmosphere. TLC (petroleum ether / ethyl acetate = 1 / 1, R of compound 18) f = 0.30, product R f=0.20) and LCMS analysis showed that NA-8 was completely consumed with the formation of one major spot. The reaction mixture was quenched by adding H2O (10.0 mL) and extracted with EtOAc (10.0 mL x 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by HPLC using a 10 micron Kromasil C18 column 100 mm x 40 mm. The mobile phase consisted of water (0.1% TFA) and ACN, a gradient of ACN from 32% to 54% over 10 min. The collected fractions were concentrated by lyophilization. The product was dissolved in aqueous HCl (0.5%, 10.0 mL) and MeCN (2.00 mL), then the liquid was concentrated again by lyophilization. The title compound (36.5 mg, 87.9 μmol, 25.1% yield, 96.6% purity) was obtained as a white solid. 1 H NMR: CDCl3400MHz, δ:7.82(d,J=7.82 Hz,1H),7.46-7.51(m,2H),7.29-7.43(m,3H),7.21-7.25(m,1H),7.10(d,J=8.80 Hz,1H),6.87(s,1H),5.67(s,2H),5.30(s,2H),3.77(m,J=5.9 Hz,2H),2.57(m,J=6.8 Hz,2H),2.18(s,3H),1.84(m,J=6.4 Hz,2H),1.26(s,2H).LCMS:(M+H + ).402.1, calculated value 401.17.

[0185] Intermediate compound 18 was prepared as follows.

[0186] a. 5-(2-bromophenyl)pent-4-yn-1-ol (17) TIFF2024527604000047.tif29170

[0187] A mixture of compound 16 (2.50 g, 8.84 mmol, 1.14 mL), CuI (100 mg, 530 μmol) and Pd(PPh3)2Cl2 (186 mg, 265 μmol) in Et3N (20.0 mL) was stirred at 25 °C under N2 for 30 min. Then compound 17A (892 mg, 10.6 mmol) was added and the mixture was stirred at 60 °C under Ar atmosphere for 3 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R of compound 16 was confirmed. f = 0.7, product R f =0.2) indicated that compound 16 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (20.0 mL) and extracted with EtOAc (20.0 mL x 3). The combined organic layers were washed with brine (20.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound 17 (1.71 g, 7.15 mmol, 80.9% yield) was obtained as a white solid.

[0188] b. 5-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pent-4-yn-1-ol (18) TIFF2024527604000048.tif40170

[0189] A mixture of compound 17 (1.00 g, 4.18 mmol), KOAc (821 mg, 8.36 mmol) and compound 15A (1.27 g, 5.02 mmol) in DMSO (10.0 mL) was degassed and purged with N 3 times, then Pd(dppf)Cl 2 (306 mg, 418 μmol) was added and the mixture was stirred at 120° C. for 18 h under N 2 atmosphere. TLC (petroleum ether / ethyl acetate=2 / 1, R of compound 17) f = 0.35, product R f=0.3) and HPLC analysis showed that compound 17 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (50.0 mL) and extracted with EtOAc (50.0 mL x 3). The combined organic layers were washed with brine (30.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 8 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. Compound 18 (750 mg, 2.62 mmol, 62.7% yield) was obtained as a yellow oil.

[0190] Example 7. N-(2-(3-aminophenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000049.tif24170

[0191] Compound NA-8 (500 mg, 1.55 mmol), compound 9A (425 mg, 3.10 mmol), Pd(dppf)Cl2 (114 mg, 155 μmol) and Na2CO3 (329 mg, 3.10 mmol) were suspended in HO (5.00 mL) and dioxane (25.0 mL) and heated at 95 °C for 16 h under N2 atmosphere. TLC (petroleum ether / ethyl acetate = 1 / 1, R of NA-8) f = 0.30, product R f =0.25) indicated that NA-8 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (20.0 mL) and extracted with EtOAc (20.0 mL x 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 20 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. The title compound (250 mg, 748 μmol, yield 48.2%) was obtained as a yellow solid.

[0192] Example 8. N-(2-(2-methoxyphenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)pent-3-ynamide TIFF2024527604000050.tif38170

[0193] To a solution of NA-4 (300 mg, 976 μmol) and compound 20 (192 mg, 1.95 mmol) in DMF (10.0 mL) was added EDCI (374 mg, 1.95 mmol) at 0° C. The mixture was stirred at 18° C. for 2 h. TLC (petroleum ether / ethyl acetate=1 / 1, R of NA-4 f = 0.35, product R f =0.45) and LCMS analysis showed that NA-4 was completely consumed and the main product peak was detected. The reaction mixture was quenched by adding HO (10.0 mL) and extracted with EtOAc (10.0 mL x 3). The combined organic layers were filtered and concentrated under reduced pressure. The residue was purified by HPLC using a 5 micron Boston Prime C18 column with an internal diameter of 150 mm x 30 mm. The mobile phase consisted of water (containing 0.05% HCl) and a gradient of ACN from 35% to 60% over 10 min. The combined product fractions were concentrated by lyophilization. The title compound (73.6 mg, 187 μmol, 19.1% yield, 98.3% purity) was obtained as a white solid. 1 H NMR:CDCl3400MHz,δ:8.36(s,1H),8.17(d,J=7.0 Hz,1H),7.79(s,1H),7.44(m,J=7.9 Hz,2H),7.00-7.08(m,2H),6.98-7.16(m,1H),6.81(s,1H),6.11(s,2H),5.35(s,2H),3.98(s,3H),3.34(d,J=2.19 Hz,2H),1.95(m,J=2.41 Hz,3H).LCMS:(M+H + ):388.15, calculated value 387.16.

[0194] The intermediate compound NA-4 was prepared as follows.

[0195] a. 2-(2-Methoxyphenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-amine (NA-4) TIFF2024527604000051.tif38170

[0196] A mixture of compound 6 (4.00 g, 14.3 mmol), compound 4A (2.60 g, 17.1 mmol), Pd(dppf)Cl2 (2.09 g, 2.86 mmol) and K2CO3 (3.95 g, 28.6 mmol) in DME (40.0 mL) and H2O (4.00 mL) was degassed and purged with N2 three times. The mixture was stirred at 95 °C under N2 atmosphere for 12 h. TLC (petroleum ether / ethyl acetate = 1 / 1, R of compound 6) was confirmed. f = 0.43, product R f =0.24) indicated the reaction was complete. The mixture was poured into H2O (100 mL) and extracted with EtOAc (50.0 mL × 3). The combined organic layers were washed with brine (20.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). Compound NA-4 (3.00 g, 9.63 mmol, yield 67.4%, purity 98.6%) was obtained as a yellow solid. 1 H NMR:DMSO 400MHz,δ:7.84(m,J=7.62 Hz,1H),7.28(m,J=8.4 Hz,1H),7.07(m,J=7.82 Hz,1H),6.94-7.00(m,2H),6.72(s,1H),6.21(m,J=8.12 Hz,1H),6.16(m,J=2.4 Hz,1H),5.40(s,2H),5.24(s,2H),5.15(s,2H),3.84(s,3H),3.32(s,2H).LCMS:(M+H + ):308.05, calculated value 307.13.

[0197] Example 9. N-(2-(4-amino-2-(5-hydroxypent-1-yn-1-yl)phenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000052.tif33170

[0198] A mixture of compound 26 (467 mg, 1.55 mmol), compound NA-8 (250 mg, 776 μmol) and Na2CO3 (165 mg, 1.55 mmol) in HO (2.50 mL) and dioxane (12.5 mL) was degassed and purged with N2 three times, then Pd(dppf)Cl2 (56.8 mg, 77.6 μmol) was added and the mixture was stirred at 90 °C under N2 atmosphere for 9 h. TLC (ethyl acetate, starting material: R f =0.7, product: R f =0.3) and LCMS analysis showed that compound 26 was completely consumed and about 22.4% of the desired product was detected. The reaction mixture was poured into water (10.0 mL), extracted with ethyl acetate (15.0 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 30 / 1 to 1 / 3). Compound 27 (100 mg, 19.3% yield) was obtained as a brown solid.

[0199] Intermediate 26 was prepared as follows.

[0200] a. 1-Bromo-2-iodo-4-nitrobenzene (23) TIFF2024527604000053.tif24170

[0201] To a solution of compound 22 (10.0 g, 49.5 mmol) in CF3SO3H (22.0 mL) was added NIS (11.1 g, 49.5 mmol) in portions at 0° C. After addition, the mixture was stirred at 25° C. for 2 h. TLC (petroleum ether, starting material: R f =0.5, product: R f=0.45), indicating that compound 22 was completely consumed. The reaction mixture was quenched with ice water (20.0 mL) and extracted with DCM (25.0 mL x 3). The combined organic extracts were washed with 10% aqueous sodium sulfite solution (20.0 mL) and water (10.0 mL), dried over Na2SO4, and evaporated under reduced pressure to give compound 23 (11.5 g, 70.9% yield) as a brown solid, which was used in the next step without further purification.

[0202] b. 4-Bromo-3-iodoaniline (24) TIFF2024527604000054.tif24170

[0203] To a solution of compound 23 (11.6 g, 35.4 mmol) in EtOH (15.0 mL) and HO (15.0 mL), HCl (12 M, 4.42 mL) and Fe (5.93 g, 106 mmol) were added dropwise at 90° C. The resulting mixture was stirred at 90° C. for 2 h. TLC (petroleum ether / ethyl acetate=3 / 1, starting material: R f =0.5, product: R f =0.3) indicated that compound 23 was completely consumed. The residue was poured into water (10.0 mL). The aqueous phase was extracted with ethyl acetate (30.0 mL×3). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give compound 24 (10.5 g, 99.6% yield) as a brown oil, which was used in the next step without further purification.

[0204] c. 5-(5-amino-2-bromophenyl)pent-4-yn-1-ol (25) TIFF2024527604000055.tif32170

[0205] To a solution of compound 24 (5.50 g, 18.5 mmol) in ACN (20.0 mL) was added compound 14A (1.86 g, 22.2 mmol) and TEA (3.74 g, 36.9 mmol, 5.14 mL). The mixture was degassed with Ar, then CuI (70.3 mg, 369 μmol) and Pd(PPh3)2Cl2 (259 mg, 369 μmol) were added. The reaction was heated at 60° C. for 5 h. TLC (petroleum ether / ethyl acetate=3 / 1, starting material: R f =0.3, product: R f =0.2) indicated that compound 24 was completely consumed. The reaction mixture was concentrated under reduced pressure to remove ACN. The residue was diluted with H2O (10.0 mL), extracted with EtOAc (10.0 mL x 3), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 2 / 1). Compound 25 (3.40 g, 72.5% yield) was obtained as a brown solid.

[0206] d. 5-(5-amino-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)pent-4-yn-1-ol (26) TIFF2024527604000056.tif35170

[0207] A mixture of compound 25 (4.40 g, 17.3 mmol), compound 15A (8.79 g, 34.6 mmol) and KOAc (5.10 g, 51.9 mmol) in DMSO (25.0 mL) was degassed and purged with N2 three times. Pd(dppf)Cl2·CH2Cl2 (1.41 g, 1.73 mmol) was added, and the mixture was then stirred at 90 °C under N2 atmosphere for 2 h. TLC (petroleum ether / ethyl acetate = 1 / 1, starting material: R f =0.4, product: R f=0.45) indicated that compound 25 was completely consumed. LCMS analysis indicated that compound 25 was completely consumed, with about 21.6% of the desired mass being detected. The residue was diluted with H2O (20.0 mL), extracted with EtOAc (20.0 mL x 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give compound 26 (660 mg, 12.7% yield) as a brown oil. 1 H NMR:CDCl3-400MHz,δ:7.50(d,J=8.0 Hz,1 H),6.65(d,J=2.4 Hz,1 H),6.50(dd,J=8.0,2.4 Hz,1 H),4.05(q,J=7.2 Hz,1 H),3.80(t,J=6.0 Hz,2 H),2.50(t,J=6.4 Hz,2 H),1.97(s,1 H),1.79(q,J=6.4 Hz,3 H),1.47(s,3 H),1.26(s,12 H).

[0208] Example 10. N-(2-(4-hydroxy-2-(5-hydroxypent-1-yn-1-yl)phenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000057.tif38170

[0209] To a solution of 3-(5-hydroxypent-1-yn-1-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (703 mg, 2.33 mmol) in dioxane (25.0 mL) and HO (5.00 mL), compound NA-8 (500 mg, 1.55 mmol), NaCO (329 mg, 3.10 mmol) and Pd(dppf)Cl (114 mg, 155 μmol) were added at 25 °C, the reaction mixture was degassed and purged with N three times, and then the mixture was stirred under N atmosphere at 90 °C for 12 h. LCMS analysis showed that compound 27 was completely consumed and one main peak of the desired product was detected. The reaction mixture was diluted with HO (7.00 mL) and extracted with EtOAc (5.00 mL × 3). The combined organic layers were washed with brine (5.00 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 0 / 1). The title compound (160 mg, yield 24.6%) was obtained as a pale yellow solid.

[0210] The intermediate 3-(5-hydroxypent-1-yn-1-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol was prepared as follows.

[0211] a. 4-Bromo-3-iodophenol

[0212] To a solution of 2-iodophenol (20.0 g, 90.9 mmol) was added Br2 (18.2 g, 114 mmol, 5.86 mL) in AcOH (120 mL) at 15 °C. The mixture was stirred at 25 °C for 2 h. HPLC analysis showed that compound 24 was completely consumed. The reaction mixture was stirred at 25 °C, a saturated solution of Na2S2O3 (100 mL) was added, which was then neutralized with NaHCO3 (40.0 mL) and the layers were separated. The aqueous layer was extracted with DCM (100 mL x 2). The combined organic layers were washed with water, dried over Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 1 / 1). 4-Bromo-3-iodophenol (23.0 g, 84.6% yield) was obtained as a white solid. 1 H NMR: CDCl3-400MHz, δ:7.42(d,J=8.8 Hz,1H),7.37(t,J=2.8 Hz,1H),6.73-6.70(m,1H).

[0213] b. 4-Bromo-3-(5-hydroxypent-1-yn-1-yl)phenol

[0214] To a solution of compound 4-bromo-3-iodophenol (23.0 g, 77.0 mmol) in ACN (115 mL) was added compound 5-hydroxypentyne (9.06 g, 108 mmol) and TEA (15.6 g, 154 mmol, 21.4 mL). The mixture was degassed with N2, then Pd(PPh3)2Cl2 (1.08 g, 1.54 mmol) and CuI (586 mg, 3.08 mmol) were added. The reaction was heated at 70° C. for 2 hours. TLC (ethyl acetate, product: R f=0.39), indicating that 4-bromo-3-iodophenol was completely consumed and one new spot was formed. The reaction mixture was diluted with H2O (70.0 mL) and extracted with ethyl acetate (25.0 mL × 3). The combined organic layers were washed with brine (20.0 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 0 / 1). 4-Bromo-3-(5-hydroxypent-1-yn-1-yl)phenol (16.0 g, 81.5% yield) was obtained as a white solid. 1 H NMR:CDCl3-400MHz,δ:7.35(d,J=8.8 Hz,1H),6.9(d,J=2.8 Hz,1H),6.67-6.64(m,1H),3.89(t,J=6.4 Hz,2H),2.59(t,J=6.8 Hz,2H),1.93-1.86(m,2H).3

[0215] c. 3-(5-hydroxypent-1-yn-1-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol

[0216] To a solution of 4-bromo-3-(5-hydroxypent-1-yn-1-yl)phenol (16.0 g, 62.7 mmol) in DMSO (160 mL) was added compound 15A (31.9 g, 125 mmol), Pd(dppf)Cl2·CH2Cl2 (5.12 g, 6.27 mmol) and KOAc (18.5 g, 188 mmol) at 25 °C. The reaction mixture was degassed and purged with N2 three times, then stirred at 90 °C under N2 atmosphere for 2 h. LCMS analysis showed that 4-bromo-3-(5-hydroxypent-1-yn-1-yl)phenol was completely consumed and one peak of the desired product was detected. The reaction mixture was diluted with H2O (70.0 mL) and extracted with ethyl acetate (25.0 mL × 3). The combined organic layers were washed with brine (20.0 mL x 2), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1). 3-(5-hydroxypent-1-yn-1-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (10.4 g, 49.8% yield, 90.8% purity) was obtained as a brown oil. 1 H NMR:DMSO-d6400MHz,δ:7.45(d,J=8.0 Hz,1H),6.74-6.68(m,2H),3.58-3.54(m,2H),2.43(t,J=6.8 Hz,2H),1.73-1.65(m,2H),1.08(s,12H).LCMS:(M+H + ):303.1, calculated value 302.17.

[0217] Example 12. 2-(4-(2-phenyl-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)-1H-pyrazol-1-yl)acetonitrile TIFF2024527604000058.tif37170

[0218] To a vial containing a solution of 28 (0.350mmol, 1.00eq), 29 (0.525mmol, 1.50eq) in dioxane (3.0mL) was added K3PO4 (2.0M, 350ul) and Pd-118 (0.018mmol, 0.05eq). The mixture was degassed and purged with N2, then the mixture was stirred at 100°C under N2 atmosphere for 16 hours. The reaction mixture was concentrated under reduced pressure to remove dioxane. 1.0mL of water was added to the reaction mixture and extracted with EtOAc (3mL x 3). The organic layer was collected and the solvent was removed by Speedvac to give a residue. The residue was purified by preparative HPLC (Xtimate C18 150*25mm*5um column under formic acid condition (B)) to give the desired product Example 12 as a white solid. 1 H NMR(400 MHz,DMSO-d6)δ 8.28(s,1H),8.06(s,1H),7.75-7.80(m,2H),7.37-7.42(m,3H),7.26-7.32(m,1H) ,7.18-7.25(m,2H),6.80(s,1H),5.65(s,2H),5.49(s,2H),5.39(s,2H).LCMS:(M+H + ):368, calculated value 367.

[0219] This general procedure was used to prepare Examples 11-17. Condition A = Prep HPLC under basic conditions; Condition B = Prep HPLC in formic acid; Condition C = Prep TLC. TIFF2024527604000059.tif234170TIFF2024527604000060.tif27170

[0220] a. Synthesis of 7-bromo-2-phenyl-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepane (34) TIFF2024527604000061.tif81170

[0221] b. Methyl 1-(4-bromo-2-fluorobenzyl)-3-phenyl-1H-pyrazole-5-carboxylate (32) TIFF2024527604000062.tif37170

[0222] To a solution of compound 30 (36.1 g, 134.8 mmol) in ACN (300 mL), CsF (41.0 g, 269.8 mmol) and compound 31 (30.0 g, 148.4 mmol) were added. The mixture was stirred at 25° C. for 5 h. TLC (petroleum ether / ethyl acetate=3 / 1) showed the reaction was incomplete. Filtered to obtain a solid, which was added to water (200 mL), extracted with EtOAc (300 mL×3), washed with brine (200 mL×3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain compound 32 (41.0 g, 78.1% yield) as a white solid. 1H NMR (300 MHz, CDCl3): δ 7.86-7.84(m,2H),7.59(dd,J=9.9,2.1 Hz,1H),7.48-7.35(m,5H),6.94(t,J=8.1 Hz,1H),3.84(s,3H).LCMS:389.0,391.0([M+H] + ).

[0223] c. (1-(4-bromo-2-fluorobenzyl)-3-phenyl-1H-pyrazol-5-yl)methanol (33) TIFF2024527604000063.tif37170

[0224] To a mixture of compound 32 (10.0 g, 25.7 mmol) in THF (100 mL) was added DIBAL-H (1.5 M, 34.0 mL) at 0° C. The mixture was stirred at 15° C. for 1 h under N2 atmosphere. TLC (petroleum ether / ethyl acetate=5 / 1) showed that compound 32 was completely consumed. The reaction mixture was quenched by adding potassium sodium tartrate solution (30 mL). Extraction with EtOAc (100 mL×3), washing with brine (30 mL×3), drying over anhydrous Na2SO4, filtering, concentrating under reduced pressure, and recrystallization in MTBE gave compound 33 (8.0 g, 86.0% yield) as a white solid. 1H NMR(300 MHz,DMSO-d6):δ 7.76-7.74(m,2H),7.59(dd,J=9.6,1.8 Hz,1H),7.40-7.35(m,3H),7.31-7.25(m,1H),6.99(t,J=8.1 Hz,1H),5.44-5.40(m,3H),4.53(d,J=5.4 Hz,2H).LCMS:361.0,363.0([M+H] + ).

[0225] d. 7-Bromo-2-phenyl-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepane (34) TIFF2024527604000064.tif37170

[0226] To a suspension of NaH (5.7 g, 60%, 141.7 mmol) in PhMe (1.2 L), compound 33 (23.3 g, 64.5 mmol) and HMPA (230 mL) were added, the resulting solution was heated to 95° C. and the temperature was maintained overnight, after which A4 was completely consumed as monitored by LCMS, the mixture was cooled to room temperature, quenched with water (250 mL), extracted with EtOAc (300 mL×3), washed with water (50 mL), brine (50 mL×4), dried over Na2SO4, concentrated to dryness, and the residue was recrystallized with EtOAc to give compound 34 (16.0, 72.7%) as an off-white solid. 1 H NMR(400MHz,DMSO-d6):δ 7.84-7.76(m,2H),7.41-7.36(m,3H),7.32-7.28(m,1H),7.20-7.14(m,2H),6.82(s,1H),5.67(s,2H),5.41(s,2H).LCMS:341.0,343.1([M+H] + ).

[0227] Example 21. N-Cyclopentyl-2-phenyl-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-amine TIFF2024527604000065.tif38170

[0228] Aliphatic amines: To a vial containing a solution of compound 34 (34.12 mg, 0.100 mmol, 1.0 equiv.) and compound 35 (1.0 equiv., 0.150 mmol) in 1,4-dioxane (1.00 mL), Cs2CO3 (97.5 mg, 0.300 mmol), BrettPhos (2.68 mg, 0.01 mmol) and BrettPhos-PdG3 (4.53 mg, 0.01 mmol) were added under the protection of N2. The verre was capped and the mixture was stirred at 100 °C for 16 h. The mixture was concentrated under reduced pressure. The residue was diluted with 1.0 mL of H2O and extracted with EtOAc (1.50*3 mL). The organic layer was collected and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product Example 21 (27.3 mg, 79%) as a white solid. 1 H NMR(400 MHz,DMSO-d6):7.73-7.79(m,2H),7.35-7.41(m,2H),7.25-7.30(m,1H),7.03(d,J=8.44 Hz,1H),6.71(s,1H),6.22(dd,J=2.26,8.25 Hz,1H),6.13(d,J=2.20 Hz,1H),5.68(d,J=6.60 Hz,1H),5.41(s,2H),5.26(s,2H),3.63(sxt,J=6.21 Hz,1H),1.82-1.92(m,2H),1.58-1.68(m,2H),1.49-1.55(m,2H),1.33-1.43(m,2H).LCMS:346([M+H] + ). This general procedure was used to prepare Examples 21, 23, 26, 29, 30, 37-42. Condition A = Prep HPLC under basic conditions; Condition B = Prep HPLC under formic acid; Condition C = Prep TLC.

[0229] Example 28: N-(5-chloro-1H-pyrazol-4-yl)-2-phenyl-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-amine TIFF2024527604000066.tif38170

[0230] Aniline / Amino Heterocycle To a vial containing a solution of compound 34 (34.12 mg, 0.100 mmol, 1.0 equiv.) and compound 36 (1.0 equiv., 0.150 mmol) in tert-butyl methyl ether (1.00 mL), t-BuONa (19.6 mg, 0.200 mmol), BrettPhos (2.68 mg, 0.010 mmol), and BrettPhos-Pd G3 (4.53 mg, 0.010 mmol) were added under the protection of N2. The veil was capped and the mixture was heated at 150° C. for 5 h. The reaction gave the desired product as confirmed by LCMS. The mixture was concentrated under reduced pressure. The residue was diluted with 1.0 mL of H2O and extracted with EtOAc (1.50*3 mL). The organic layer was collected and concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product Example 28 (11.6 mg, 31%) as a white solid. 1 H NMR(400 MHz,DMSO-d6):(br s,1H),7.82(d,J=1.59 Hz,1H),7.73-7.79(m,2H),7.34-7.41(m,3H),7.25-7.31(m,1H),7.10(d,J=8.31 Hz,1H),6.73(s,1H),6.29(dd,J=2.32,8.31 Hz,1H),6.16(d,J=2.32 Hz,1H),5.46(s,2H),5.27(s,2H).LCMS:378([M+H] + ).

[0231] This general procedure was used to prepare Examples 18-20, 22, 24, 25, 27, 28, 31-36. Condition A = Prep HPLC under basic conditions; Condition B = Prep HPLC under formic acid; Condition C = Prep TLC. TIFF2024527604000067.tif248170TIFF2024527604000068.tif254170TIFF2024527604000069.tif252170TIFF2024527604000070.tif104170

[0232] Example 67: N-(2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000071.tif34170

[0233] To a vial containing a solution of NA-8 (0.120 mmol, 1.00 equiv), compound 37 (0.180 mmol, 1.50 equiv) in dioxane (1.0 mL), K3PO4 (2M, 120 uL) and PdCl2(dtbpf) (0.006 mmol, 0.05 equiv) were added. The mixture was degassed and purged with N2, then the mixture was stirred at 100 °C under N2 atmosphere for 16 h. The reaction mixture was concentrated under reduced pressure to remove dioxane. 1.0 mL of water was added to the reaction mixture and extracted with EtOAc (1.0 mL*2). The organic layer was collected and the solvent was removed by Speedvac to give a residue. The residue was purified by preparative HPLC to give Example 67 (14.1 mg, 33%) as a white solid. 1 H NMR(400 MHz,DMSO-d6)δ 9.96(s,1H),8.52(s,1H),8.07(s,1H),7.65-7.98(m,1H),7.23-7.30(m,2H),7.13(dd,J=1.69,8.32 Hz,1H),6.62(s,1H),5.54(s,2H),5.33(s,2H),2.01(s,3H).LCMS:360([M+H] + ).

[0234] This general procedure was used to make Examples 43-45, 47-50, 42-56, 58-62, 64-72, 74, 76-83, 85, 89-90, 92-96, 100, 103, 106-107. Condition A = Prep HPLC under basic conditions; Condition B = Prep HPLC under formic acid; Condition C = Prep TLC.

[0235] Example 46: N-(2-(4-fluoro-2-(hydroxymethyl)phenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000072.tif37170

[0236] To a vial containing a solution of NA-8 (0.120 mmol, 1.00 equiv), compound 38 (0.180 mmol, 1.50 equiv) in dioxane (1.0 mL), K3PO4 (2M, 120 uL) and Xphos Pd G3 (0.006 mmol, 0.05 equiv) were added. The mixture was degassed and purged with N2, then the mixture was stirred at 100° C. under N2 atmosphere for 16 h. The reaction mixture was concentrated under reduced pressure to remove dioxane. 1.0 mL of water was added to the reaction mixture and extracted with EtOAc (1.0 mL*2). The organic layer was collected and the solvent was removed by Speedvac to give a residue. The residue was purified by preparative HPLC to give Example 46 (26.4 mg, 60%) as a white solid. 1 H NMR(400 MHz,DMSO-d6)δ 9.97(s,1H),7.59(dd,J=5.94,8.57 Hz,1H),7.33-7.40(m,1H),7.25-7.32(m,2H),7.04-7.18(m,2H),6.65(s,1H),5.58(s,2H),5.31-5.39(m,3H),4.68(d,J=5.75 Hz,2H),2.02(s,3H).LCMS:368([M+H] + ). This general procedure was used to make Examples 46, 51, 57, 63, 73, 75, 84, 86-88, 91, 97-99, 101-102, 104-105. Condition A = Prep HPLC under basic conditions; Condition B = Prep HPLC under formic acid; Condition C = Prep TLC.

[0237] Example 125: N-(2-(1-(trifluoromethyl)-1H-pyrazol-4-yl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide TIFF2024527604000073.tif33170

[0238] To a vial containing a solution of NA-8 (0.30 mmol, 1.00 equiv.), compound 39 (0.30 mmol, 1.0 equiv.) in dioxane (0.15 M) and water (0.15 M), K3PO4 (3 mmol, 10 equiv.) and Xphos Pd G3 (0.015 mmol, 0.05 equiv.) were added. The mixture was degassed and purged with N2, and then the mixture was stirred at 100° C. under N2 atmosphere for 16 h. The product was precipitated after diluting the reaction mixture with water and filtering. The crude product was purified by preparative HPLC to give Example 125 (59.6 mg, 53%) as a white solid. 1 H NMR(400 MHz,DMSO-d6)δ 9.95(s,1H),8.79(s,1H),8.24(t,J=0.9 Hz,1H),7.31-7.23(m,2H),7.13(dd,J=8.3,2.1 Hz,1H),6.66(s,1H),5.55(s,2H),5.35(s,2H),2.02(s,3H).LCMS:378.1([M+H] + ). This general procedure was used to make Examples 108-129. Condition A = Prep HPLC under basic conditions; Condition B = Prep HPLC under formic acid; Condition C = Prep TLC. TIFF2024527604000074.tif232170TIFF2024527604000075.tif234170TIFF2024527604000076.tif252170TIFF2024527604000077.tif252170 TIFF2024527604000078.tif254170TIFF2024527604000079.tif248170TIFF2024527604000080.tif242170TIFF2024527604000081.tif236170 TIFF2024527604000082.tif242170TIFF2024527604000083.tif253170TIFF2024527604000084.tif253170TIFF2024527604000085.tif253170 TIFF2024527604000086.tif254170TIFF2024527604000087.tif255170TIFF2024527604000088.tif255170TIFF2024527604000089.tif237170

[0239] Assay Example 1. LDC7559 does not inhibit GSDMD LDC7559 was reported to block the pore-forming activity of GSDMD (Sollberger et al., 2018). We tested its ability to inhibit GSDMD-dependent pyroptosis in primary human monocytes and human monocytic THP-1 cells (Figure 1A-Figure 1C). Lactate dehydrogenase (LDH) release was used to monitor cell death, and LDC7559 did not alter GSDMD-dependent pyroptosis induced by nigericin, cytoplasmic LPS, or cytoplasmic poly(deoxyadenosine-deoxythymidine) [poly(dA-dT)] (Figure 1A). GSDMD-dependent release of interleukin-1β (IL-1β) from monocytes in response to nigericin or poly(dA-dT) also occurred normally in the presence of LDC7559 (Figure 1B). Western blotting confirmed that LDC7559 did not prevent LPS-induced cleavage of GSDMD (Figure 1C). Thus, LDC7559 did not inhibit cleavage of recombinant GSDMD by caspase-4 (Figure 1D), nor did it prevent GSDMD pore-forming fragments from permeabilizing liposomes (Figure 1E). In contrast, inhibition of GSDMD by disulfiram (Hu et al., 2020) prevented liposome permeabilization (Figure 1E). It was revealed that LDC7559 does not inhibit GSDMD.

[0240] Assay Example 2. Quantification of NETosis using SYTOX Green. Semi-automated analysis of both phase and fluorescence images of SYTOX Green stained neutrophils used MATLAB algorithms. Total cells were counted by scoring phase standard deviation and radial symmetry. NETosis was classified into cells by measuring DNA area in fluorescence images and comparing with a selected internal training set for each experiment. Unstimulated cells and PMA stimulated pyocyanin-inhibited cells served as negative controls. PMA stimulated cells served as positive controls. NET forming cells are presented as a percentage of total cells. Data for representative compounds of the present invention are provided in the following table. TIFF2024527604000090.tif155170

[0241] Semi-automated analysis of both phase and fluorescence images of SYTOX Green stained neutrophils used MATLAB algorithms. Total cells were counted by scoring phase standard deviation and radial symmetry. NETosis was classified into cells by measuring DNA area in fluorescence images and comparing with a selected internal training set for each experiment. Unstimulated cells and PMA stimulated pyocyanin-inhibited cells served as negative controls. PMA stimulated cells served as positive controls. NET forming cells are presented as a percentage of total cells. Data at 5uM concentration are shown in the table below. TIFF2024527604000091.tif243170TIFF2024527604000092.tif255170TIFF20245276040 00093.tif243170TIFF2024527604000094.tif254170TIFF2024527604000095.tif248170 TIFF2024527604000096.tif243170TIFF2024527604000097.tif236170TIFF20245276040 00098.tif255170TIFF2024527604000099.tif253170TIFF2024527604000100.tif214170

[0242] The activity of the compounds and salts may also be assessed in biochemical assays that measure PFKL, PFKM and PFKP activity.

[0243] Assay Example 3. ROS Detection Assay ROS in neutrophils was measured using the ROS-Glo bioluminescence assay (Promega) for direct detection of H2O2. Neutrophils were isolated from fresh human blood using a biphasic Histopaque gradient as described herein. Cells (100uL / well) were seeded in 96-well plates at a density of 200,000 cells / mL in high glucose DMEM medium. Cells were pre-incubated with DMSO vehicle or compounds (10-point dilution curve starting at 20uM with 3-fold serial dilutions) for 30 minutes and then stimulated with 50nM PMA for 30 minutes. H2O2 substrate solution was added for 20 minutes, followed by ROS-Glo detection solution for 20 minutes, after which luminescence was recorded on a luminescence plate reader. The 50% inhibitory concentration (IC 50 ) was determined by plotting the assay output (RLU) versus the log of compound concentration using GraphPad Prism with a nonlinear sigmoid with variable slope algorithm using the default curve fitting parameters in the Prism software. TIFF2024527604000101.tif228170TIFF2024527604000102.tif242170TIFF2024527604000103.tif254170

[0244] Formulation Example 1. The following illustrate representative pharmaceutical dosage forms containing a compound of formula (I) ("Compound X") for therapeutic or prophylactic use in humans.

[0245] (i) Tablets 1 mg / tablet Compound X= 100.0 Lactose 77.5 Povidone 15.0 Croscarmellose sodium 12.0 Microcrystalline Cellulose 92.5 Magnesium stearate 3.0 300.0

[0246] (ii) Tablets 2 mg / tablet Compound X= 20.0 Microcrystalline Cellulose 410.0 Starch 50.0 Sodium starch glycolate 15.0 Magnesium stearate 5.0 500.0

[0247] (iii) Capsules mg / capsule Compound X= 10.0 Colloidal silicon dioxide 1.5 Lactose 465.5 Gelatinized starch 120.0 Magnesium stearate 3.0 600.0

[0248] (iv) Injection 1 (1mg / ml) mg / ml Compound X=(free acid form) 1.0 Disodium phosphate 12.0 Sodium phosphate monobasic 0.7 Sodium chloride 4.5 1.0N sodium hydroxide solution (pH adjustment to 7.0-7.5) Appropriate amount Add water for injection to 1mL

[0249] (v) Injection 2 (10mg / ml) mg / ml Compound X=(free acid form) 10.0 Sodium phosphate monobasic 0.3 Disodium phosphate 1.1 Polyethylene glycol 400 200.0 1.0N sodium hydroxide solution (pH adjustment to 7.0-7.5) Appropriate amount Add water for injection to 1mL

[0250] (vi) Aerosol mg / can Compound X= 20.0 Oleic acid 10.0 Trichloromonofluoromethane 5,000.0 Dichlorodifluoromethane 10,000.0 Dichlorotetrafluoroethane 5,000.0

[0251] The above formulations may be obtained by conventional procedures well known in the pharmaceutical art.

[0252] Preparation Example 1 N-(2-(2-methoxyphenyl)-4H,10H-benzo[f]pyrazolo[5,1-c][1,4]oxazepin-7-yl)acetamide (LDC7559): TIFF2024527604000104.tif29170

[0253] Compound NA-4 (200 mg, 651 μmol) was dissolved in THF (6.00 mL). EtN (132 mg, 1.30 mmol) was added, followed by acetyl chloride (61.3 mg, 781 μmol), and the mixture was stirred at 18° C. for 25 min. TLC (petroleum ether / ethyl acetate=1 / 1, R of NA-4) f = 0.50, product R f =0.30) indicated that NA-4 was completely consumed and one major spot was formed. The reaction mixture was quenched by adding H2O (15.0 mL) and extracted with EtOAc (15.0 mL x 3). The combined organic layers were washed with brine (10.0 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 0 / 1). The combined organic layers were concentrated under reduced pressure. The residue was purified by HPLC using a Waters Xbridge 150 mm x 25 mm and 5 μm particle size. The mobile phase consisted of 10 mM NH4HCO3 (aq) and ACN (gradient from 25% to 55% of ACN over 20 min). The collected fractions were concentrated by lyophilization to give compound LDC7559 (188 mg, 532 μmol, 81.7% yield, 98.8% purity) as a white powder. 1H NMR:CDCl3400MHz,δ:7.87(d,J=7.45 Hz,1H),7.27-7.37(m,3H),7.14-7.20(m,1H),6.94-7.11(m,3H),6.69(s,1H),5 .51(s,2H),5.27(s,2H),3.89(s,3H),2.16(s,3H),1.91-2.05(m,2H).LCMS:(M+H + ):350.05,calculated 349.14.

[0254] All publications, patents, and patent documents are incorporated herein by reference as if individually incorporated by reference. The invention has been described with reference to various specific preferred embodiments and techniques. However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the invention.

Claims

1. Formula (I): (wherein R １ is a 5- to 10-membered heteroaryl which may be substituted with -NR ａ R ｂ , or one or more groups R ｃ ; and may also be substituted; R ２ is a 6- to 10-membered aryl which may be substituted with one or more groups R ｒ ; or R ２ is a 5- to 10-membered heteroaryl which may be substituted with one or more groups R ｓ ; or R ２ is a 3- to 10-membered heterocyclic ring which may be substituted with one or more groups R ｚ ; R ａ is (C １ -C ６ ), alkyl, (C ３ -C ６ ), cycloalkyl, (C ２ -C ６ ), alkenyl, (C ２ -C ６ ), alkynyl, (C １ -C ６ ), alkanoyl, (C ３ -C ６ ), cycloalkyl(C １ -C ６ ), alkyl, (C ２ -C ６ ), alkynylcarbonyl, a 3- to 6-membered heterocyclic ring, or a 5- to 6-membered heteroaryl optionally substituted with one or more groups R ｆ ; each (C １ -C ６ ), alkyl, (C ３ -C ６ ), cycloalkyl, (C ２ -C ６ ), alkenyl, (C ２ -C ６ ), alkynyl, (C １ -C ６ ), alkanoyl, (C ３ -C ６ ), cycloalkyl(C １ -C ６ ), alkyl, (C ２ -C ６ ), alkynylcarbonyl, and the 3- to 6-membered heterocyclic ring are each independently selected from the group consisting of halo, hydroxy, cyano, (C ２ -C ６ ), alkenyl, (C ２ -C ６ ), alkynyl, C(=O)NR ｍ R ｎ ; and halo, hydroxy, cyano, -NR ｍ R ｎ ; and -C(=O)NR ｍ R ｎ ; and are optionally substituted with one or more groups independently selected from the group consisting of (C １ -C ６ optionally substituted with one or more groups independently selected from the group consisting of alkyl; R ｂ is H or (C １ -C ６ )alkyl; Each R ｃ is independently selected from the group consisting of cyano, -NR ｄ R ｅ , -C(=O)NR ｄ R ｅ , (C １ -C ６ )alkyl, (C ３ -C ６ )cycloalkyl, (C ３ -C ６ )cycloalkyl(C １ -C ６ )alkyl, (C １ -C ６ )alkoxy, and (C １ -C ６ )alkanoyl; each (C １ -C ６ )alkyl, (C ３ -C ６ )cycloalkyl, (C ３ -C ６ )cycloalkyl(C １ -C ６ )alkyl, (C １ -C ６ )alkoxy, and (C １ -C ６ )alkanoyl may be substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, and cyano; R ｄ and R ｅ are each independently selected from the group consisting of H and (C １ -C ６ )alkyl; or R ｄ and R ｅ together with the nitrogen to which they are attached form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C １ -C ６ )alkyl; Each R ｆ is selected independently from the group consisting of halo, hydroxy, cyano, -NR ｇ R ｈ , -C(=O)NR ｇ R ｈ , and may be substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, -NR ｇ R ｈ , -C(=O)NR ｇ R ｈ , and cyano; and is independently selected from the group consisting of (C １ -C ６ )alkyl; R ｇ and R ｈ are each independently selected from the group consisting of H and (C １ -C ６ )alkyl; or, R ｇ and R ｈ together with the nitrogen to which they are attached form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C １ -C ６ )alkyl; R ｍ is optionally substituted with one or more groups independently selected from the group consisting of H, or halo, hydroxy, carboxy, cyano and oxo (C １ -C ６ ) alkyl; R ｎ is optionally substituted with one or more groups independently selected from the group consisting of H, or halo, hydroxy, carboxy, cyano and oxo; (C １ -C ６ )alkyl; Each R ｒ is independently selected from the group consisting of halo, hydroxy, cyano, -NR ｔ R ｕ , -C(=O)NR ｔ R ｕ , -S(O) ２ NR ｔ R ｕ , (C １ -C ６ ), alkyl, (C ３ -C ６ ), cycloalkyl, (C １ -C ６ ), alkoxy, (C １ -C ６ ), alkylthio, -N(H)S(O) ２ R ｘ , -S(O) ２ R ｘ , (C ２ -C ６ ), alkenyl, and (C ２ -C ６ ), alkynyl; and each (C １ -C ６ ), alkyl, (C ３ -C ６ ), cycloalkyl, (C １ -C ６ ), alkoxy, (C １ -C ６ ), alkylthio, (C ２ -C ６ ), alkenyl, and (C ２ -C ６ ), alkynyl may be substituted with one or more groups independently selected from the group consisting of halo, hydroxy, carboxy, -NR ｔ R ｕ , -C(=O)NR ｔ R ｕ , -S(O) ２ NR ｔ R ｕ , -S(O) ２ R ｘ , and cyano; Each R ｓ is halo, cyano, -NR ｖ R ｗ , -C(=O)NR ｖ R ｗ , -S(O) ２ N.R. ｖ R ｗ , (C １ -C ６ ) alkyl, (C ３ -C ６ ) cycloalkyl, (C １ -C ６ ) alkoxy, (C １ -C ６ ) alkanoyl, (C １ -C ６ ) alkylthio, 3- to 6-membered heterocycle, and -S(O) ２ R ｙ Each (C １ -C ６ ) alkyl, (C ３ -C ６ ) cycloalkyl, (C １ -C ６ ) alkoxy, (C １ -C ６ ) alkanoyl, 3- to 6-membered heterocycle, and (C １ -C ６ ) Alkylthio is halo, hydroxy, carboxy, -NR ｔ R ｕ , -C(=O)NR ｔ R ｕ , S(O) ２ N.R. ｖ R ｗ , -S(O) ２ R ｙ and cyano; R ｔ and R ｕ are each independently selected from the group consisting of H, (C １ -C ６ )alkyl, (C １ -C ６ )alkanoyl, and (C ２ -C ６ )alkynylcarbonyl; or R ｔ and R ｕ together with the nitrogen to which they are attached form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C １ -C ６ )alkyl; R ｖ and R ｗ are each independently selected from the group consisting of H, (C １ -C ６ )alkyl, and (C １ -C ６ )alkanoyl; or R ｖ and R ｗ , together with the nitrogen to which they are attached, form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C １ -C ６ )alkyl; R ｘ is optionally substituted with one or more groups independently selected from the group consisting of H, or halo, hydroxy, carboxy, cyano and oxo (C １ -C ６ )alkyl; R ｙ is optionally substituted with one or more groups independently selected from the group consisting of H, or halo, hydroxy, carboxy, cyano and oxo; (C １ -C ６ )alkyl; Each R ｚ is optionally substituted with one or more groups independently selected from the group consisting of oxo, halo, hydroxy, and halo, hydroxy, carboxy, -NR ｔ R ｕ , -C(=O)NR ｔ R ｕ , S(O) ２ NR ｖ R ｗ , -S(O) ２ R ｙ , cyano, and oxo, and is a compound independently selected from the group consisting of (C １ -C ６ ) alkyl), provided that the compound is not the following: a compound, or a prodrug thereof, or a pharmaceutically acceptable salt thereof).

2. R １ is a 5- to 10-membered heteroaryl which may be substituted with -NR ａ R ｂ , or one or more groups R ｃ ; R ２ is phenyl which may be substituted with one or more groups R ｒ ; or R ２ is 5- to 9-membered heteroaryl which may be substituted with one or more groups R ｓ ; or R ２ is a 9-membered heterocyclic ring which may be substituted with one or more groups R ｚ ; R ａ However, (C １ -C ６ ) alkyl, (C ３ -C ６ ) cycloalkyl, (C ２ -C ６ ) alkynyl, (C １ -C ６ ) alkanoyl, (C ２ -C ６ ) alkynylcarbonyl, a 3- to 6-membered heterocycle, or one or more groups R ｆ is a 5-membered heteroaryl optionally substituted with １ -C ６ ) alkyl and (C ３ -C ６ ) Cycloalkyl is halo, cyano, (C ２ -C ６ ) alkynyl, C(=O)NR ｍ R ｎ and optionally substituted with one or more hydroxy (C １ -C ６ ) alkyl; R ｂ is H or (C １ -C ６ )alkyl; Each R ｃ is independently selected from the group consisting of cyano, -NR ｄ R ｅ , and (C １ -C ６ )alkyl which may be substituted with one or more cyanos; R ｄ and R ｅ are each H; Each R ｆ is selected independently from the group consisting of halo, hydroxy, cyano, -C(=O)NR ｇ R ｈ , and may be substituted with one or more groups independently selected from the group consisting of halo, hydroxy and carboxy, (C １ -C ６ )alkyl; R ｇ and R ｈ are each H; R ｍ is H; R ｎ is H; Each R ｒ is independently selected from the group consisting of halo, hydroxy, cyano, -NR ｔ R ｕ , -C(=O)NR ｔ R ｕ , -S(O) ２ NR ｔ R ｕ , (C １ -C ６ )alkyl, (C ３ -C ６ )cycloalkyl, (C １ -C ６ )alkoxy, (C １ -C ６ )alkylthio, -N(H)S(O) ２ R ｘ , -S(O) ２ R ｘ , and (C ２ -C ６ )alkynyl, and each (C １ -C ６ )alkyl and (C ２ -C ６ )alkynyl may be substituted with one or more groups independently selected from the group consisting of hydroxy, -NR ｔ R ｕ , -C(=O)NR ｔ R ｕ , and cyano; Each R ｓ is independently selected from the group consisting of halo, cyano, -NR ｖ R ｗ , -C(=O)NR ｖ R ｗ , (C １ -C ６ )alkyl, (C ３ -C ６ )cycloalkyl, (C １ -C ６ )alkoxy, (C １ -C ６ )alkanoyl, (C １ -C ６ )alkylthio, a 3- to 6-membered heterocyclic ring, and -S(O) ２ R ｙ , and each (C １ -C ６ )alkyl may be substituted with one or more groups independently selected from the group consisting of halo and -NR ｔ R ｕ ; R ｔ and R ｕ are each independently selected from the group consisting of H, (C １ -C ６ )alkyl, (C １ -C ６ )alkanoyl, and (C ２ -C ６ )alkynylcarbonyl; or R ｔ and R ｕ together with the nitrogen to which they are attached form a 3- to 6-membered heterocyclic ring optionally substituted with one or more groups independently selected from the group consisting of halo and (C １ -C ６ )alkyl; R ｖ and R ｗ are each independently selected from the group consisting of H, (C １ -C ６ )alkyl, and (C １ -C ６ )alkanoyl; R ｘ is (C １ -C ６ ) alkyl; R ｙ is (C １ -C ６ ) alkyl; Each R ｚ is independently selected from the group consisting of oxo and (C １ -C ６ ) alkyl, the compound according to claim 1, or a prodrug thereof, or a pharmaceutically acceptable salt thereof.

3. R ａ The compound, prodrug, or pharmaceutically acceptable salt according to claim 1 or 2, wherein R is acetyl.

4. R ｂ The compound, prodrug, or pharmaceutically acceptable salt according to claim 1 or 2, wherein R is H.

5. R ２ is a 5- to 9-membered heteroaryl which may be substituted with one or more groups R ｓ The compound, prodrug, or pharmaceutically acceptable salt according to claim 1 or 2, wherein

6. Each R ｓ is independently selected from the group consisting of cyano, -NR ｖ R ｗ , -C(=O)NR ｖ R ｗ , (C １ -C ６ )alkyl, (C ３ -C ６ )cycloalkyl, (C １ -C ６ )alkoxy, (C １ -C ６ )alkylthio, -S(O) ２ R ｙ , and (C １ -C ６ )alkanoyl, and each (C １ -C ６ )alkyl may be substituted with one or more groups independently selected from the group consisting of halo and -NR ｔ R ｕ . The compound, prodrug, or pharmaceutically acceptable salt according to claim 1 or 2

7. The following: The compound, prodrug, or pharmaceutically acceptable salt according to claim 1, selected from the group consisting of the following, its prodrug, and pharmaceutically acceptable salts thereof.

8. The following: The compound, prodrug, or pharmaceutically acceptable salt according to claim 1, selected from the group consisting of the following, its prodrug, and pharmaceutically acceptable salts thereof.

9. The following: The compound, prodrug, or pharmaceutically acceptable salt according to claim 1, selected from the group consisting of the following, its prodrug, and pharmaceutically acceptable salts thereof.

10. The following: The compound, prodrug, or pharmaceutically acceptable salt according to claim 1, selected from the group consisting of the following, its prodrug, and pharmaceutically acceptable salts thereof.

11. The following: The compound, prodrug, or pharmaceutically acceptable salt according to claim 1, selected from the group consisting of the following, its prodrug, and pharmaceutically acceptable salts thereof.

12. The following: A compound selected from the group consisting of... or a pharmaceutically acceptable salt thereof.

13. A pharmaceutical composition comprising the compound, prodrug, or pharmaceutically acceptable salt according to claim 1 or 2 and a pharmaceutically acceptable additive.

14. The compound of formula (I) according to claim 1 or 2, or a prodrug thereof, or a pharmaceutically acceptable salt thereof, for use in drug therapy.

15. The compound of formula (I) according to claim 1 or 2, or a prodrug thereof, or a pharmaceutically acceptable salt thereof, for the prophylactic or therapeutic treatment of a disease associated with the activity of glycolytic enzyme phosphofructokinase-1 liver type.

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