NLRP3 inhibitors

Abstract

The present invention relates to compounds useful as NOD-like receptor protein 3 (NLRP3) inflammasome pathway inhibitors, processes for preparing the compounds, pharmaceutical compositions comprising the compounds, methods of treating various diseases and disorders using the compounds, and medicaments containing them, and their use in NLRP3 mediated diseases and disorders.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to PCT Application No. PCT / US2022 / 075421, filed on August 24, 2022, and U.S. Provisional Patent Application No. 63 / 311,463, filed on February 18, 2022, the contents of which are hereby incorporated by reference in their entirety and for all purposes. Technical Field

[0003] The present invention relates to compounds that can be used as inhibitors of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway. The present invention also relates to methods for preparing the compounds, pharmaceutical compositions comprising the compounds, methods for using the compounds to treat various diseases and conditions, and drugs comprising the compounds, as well as their use in NLRP3-mediated diseases and conditions. Background Art

[0004] The term inflammasome was coined by Martinon et al. to describe a molecular platform that triggers the activation of inflammatory caspases and the processing of interleukin 1 (IL-1) family cytokines (Fabio Martinon et al., Mol Cell 10(2):417-26, 2002). Inflammasomes are part of the innate immune system. Activation of inflammasomes is initiated by the assembly of a multiprotein complex including nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs), apoptosis-associated speck-like protein (ASC) adapters containing caspase recruitment domains, and effector protease caspase 1. The assembly of the complex results in the activation of caspase 1 and the release of mature proinflammatory cytokines (e.g., IL-1β and IL-18).

[0005] Among inflammasomes, the NLR family protein containing NACHT, LRR, and PYD domains 3 (NLRP3) inflammasome has been extensively studied and found to be activated by a variety of stimuli. A recent review paper (Seungwha Paik et al., Cell Mol Immunol 18(5):1141-1160, 2021) summarizes the regulatory mechanisms of NLRP3 activation.

[0006] NLRP3 activation is triggered by a variety of infectious and non-infectious molecules, including molecular byproducts of aging, lack of exercise, and overnutrition. Once activated, it promotes the downstream production of inflammatory cytokines IL-1β and IL-18. Gain-of-function mutations in NLRP3 are associated with a variety of genetic diseases, including cold pyrin-associated periodic syndrome (CAPS). In addition, NLRP3 is involved in a variety of common I) autoimmune diseases, II) autoinflammatory diseases, III) neurodegenerative diseases, IV) cardiovascular diseases, and V) neuromuscular and muscle degenerative diseases, such as (Matthew SJ Mangan et al., Nat Rev Drug Discov 17(8):588-606, 2018; Corcoran et al., Pharmacol Rev 73(3):968-1000, 2021; Dubuisson et al., Cells 10(11):3023, 2021). Inflammasome activation has also been found in retinal pigment epithelial (RPE) cells and is considered to be one of the causes of RPE dysfunction and degeneration (Gao et al., Mediators Inflamm 2015:690243, 2015). In addition, NLRP3 activation has been associated with severe COVID-19 cases and cytokine release syndrome (CRS) caused by cell therapy and biological treatment (Tracey L Freeman and Talia H Swartz Front Immunol 11:1518, 2020; Lin et al., PLoS Pathog 6; 15(6):e1007795, 2019).

[0007] Therefore, NLRP3 inflammasome inhibitors may be used clinically as single or combined drugs as new therapeutic approaches for the treatment of these diseases. Therefore, there is a need for inhibitors of the NLRP3 inflammasome pathway to provide new and / or alternative therapies for these inflammasome-associated diseases, disorders (e.g., autoinflammatory febrile syndrome-cryopyrin-associated periodic syndrome (CAPS), sickle cell disease, chronic liver disease, nonalcoholic steatohepatitis (NASH), gout, hyperoxaluria, pseudogout (chondrocalcinosis), type I / II diabetes and related complications (e.g., nephropathy, retinopathy), fibrosis, rheumatoid arthritis, inflammatory bowel disease, asthma and allergic airway inflammation, neuroinflammation-related diseases (e.g., multiple sclerosis, brain infection, acute injury, Alzheimer's disease, Parkinson's disease, Huntington's disease), neuromuscular and muscle degenerative diseases, atherosclerosis and cardiovascular risk (e.g., cardiovascular risk reduction (CvRR), hypertension), hidradenitis suppurativa, wound healing and scar formation, and cancer (e.g., colon cancer, lung cancer, myeloproliferative neoplasms, leukemia, myelodysplastic syndrome (MDS), myelofibrosis)).

[0008] References:

[0009] Fabio Martinon, Kimberly Burns, Jürg Tschopp. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta Mol Cell 10(2):417-26, 2002.

[0010] Seungwha Paik, Jin Kyung Kim, Prashanta Silwal, Chihiro Sasakawa, Eun-Kyeong Jo. An update on the regulatory mechanisms of NLRP3 inflammasome activation Cell Mol Immunol18(5):1141-1160, 2021.

[0011] Matthew S J Mangan, Edward J Olhava, William R Roush, H Martin Seidel, Gary D Glick, Eicke Latz. Targeting the NLRP3 inflammasome in inflammatory diseases Nat Rev Drug Discov17(8):588-606, 2018

[0012] Sarah Corcoran, Reena Halai, Matthew A Coope. Pharmacological Inhibition of the Nod-Like Receptor Family Pyrin Domain Containing 3 Inflammasome with MCC950 Pharmacol Rev73(3):968-1000, 2021.

[0013] Nicolas Dubuisson, Romain Versele, María A Davis-López de Carrizosa, Camille M Selvais, Sonia M Brichard, Michel Abou-Samra. Walking down Skeletal Muscle Lane: From Inflammasome to Disease Cells 10(11):3023,2021.

[0014] Jiangyuan Gao,Ruozhou Tom Liu,Sijia Cao,Jing Z Cui,Aikun Wang,EleanorTo,Joanne AMatsubara. NLRP3 inflammasome: activation and regulation in age-related macular degeneration Mediators Inflamm 2015:690243,2015.

[0015] Tracey L Freeman and Talia H Swartz. Targeting the NLRP3 Inflammasomein Severe COVID-19Front Immunol 11:1518,2020.

[0016] Lan Lin, Lei Xu, Weihua Lv, Li Han, Yaozu Xiang, Lei Fu, Meilin Jin, Rui Zhou, Huanchun Chen, Anding Zhang. An NLRP3 inflammasome-triggered cytokinestorm contributes to Streptococcal toxic shock-like syndrome (STSLS) PLoS Pathog 6; 15(6): e1007795, 2019. Summary of the invention

[0017] The present invention provides a compound or a pharmaceutically acceptable salt thereof that inhibits the NLRP3 inflammasome pathway, and a pharmaceutical composition thereof. The present invention also provides a method for treating or preventing a disease and / or condition associated with NLRP3, comprising administering an effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0018] Various aspects of the invention are described herein.

[0019] The present application discloses compounds of formula I:

[0020]

[0021] in:

[0022] R w For hydrogen, C 1-6 Alkyl (e.g. CH3), C 2-8 Alkynyl, halogen (e.g. F, Cl), C 1-6 Alkoxy (e.g. OCH3), halo-C 1-4 Alkyl (e.g. CHF2, CF3), halo-C 1-4 Alkoxy (e.g. OCHF2, OCF3), cyano, -NH2, -N(C 1-6 Alkyl)2, mercapto, -SO2NH2, -SO2N(C 1-6 Alkyl)2, -S(=O)(C 1-6 alkyl), cycloalkyl, CHR 1a 、(C=O)R 1a , OR 1a NR 1b 、S(=O)R 1a 、S(=O)2R 1a 、O(C=O)R 1a 、(C=O)OR 1a NR 1b (C=O)R 1b 、(C=O)NR 1b 、(C=O)N(R 1b )2. NR 1b (C=O)OR 1a 、O(C=O)NR 1b 、ONR 1b (C=NR 1b )NR 1b NR 1b S(=O)2 or S(=O)2NR 1b , where each C 1-6 Alkyl, heterocycle (eg, 2,5-dihydrofuran-3-yl), heteroaryl, and aryl are optionally substituted with 1 or 2 substituents each selected from R5;

[0023] Each W is independently CH, CR' or N;

[0024] Each Q' is independently N, C or CH;

[0025] Each A is independently CH, CH2, CRa , CHR a 、CR4、CHR4、N、NH、NR a , NR4, S or O;

[0026] Each A' is independently absent, CH, CH2, CR a , CHR a 、CR4、CHR4、N、NH、NR a , NR4, S or O;

[0027] Each R a and -C(=O)R2, -OC(=O)R2, -C(=O)OR2, -OC(=O)OR2, -C(=O)N(R2)2, -OC(=O)N(R2)2, -NR2C(=O)R2, -P(=O)(R2), C(=O)R2, -C(=O)R2, -C(=O)OR2, -OC(=O)OR2, -C(=O)N(R2)2, -OC(=O)N(R2)2, -NR2C(=O)R2, -P(=O)(R2), C(=O)R2, -C(=O)R2, -C(=O)OR2, -C(=O)N(R2)2, -C(=O)R2, -C(=O) 1-4 Alkyl, (C 1-4 Alkyl)2, halo-C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-8 Cycloalkyl, C 2-7 heterocycloalkyl, aryl or monocyclic heteroaryl;

[0028] Each -- represents absence, a single bond, or a double bond;

[0029] Y is C(R 1a )2. C=O、O、NR 1b or key;

[0030] Each R 1a are independently hydrogen, halogen, hydroxyl, cyano, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino or hydroxy-C 1-4 Alkyl, C 3-10 Cycloalkyl, C 2-7 heterocycloalkyl or aryl;

[0031] Each R 1b are independently hydrogen, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl or hydroxy-C 1-4 alkyl;

[0032] Each R' is independently a heterocyclic group, a heteroaryl group, an aryl group, a C 3-8 Cycloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl (e.g. trifluoromethyl, difluoromethyl), halo-C 1-4 Alkoxy (e.g. trifluoromethoxy), hydroxy-C 1-4 Alkyl, halogen, hydroxyl or cyano,

[0033] Each Z is a heterocyclic group, a heteroaryl group, an aryl group, a C 3-10 Cycloalkyl, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, Hydroxyl-C 1-8 Alkyl, C 1-6 Alkoxy, NH(hydroxy-C 1-6 Alkyl), NH(C 1-6 alkoxy), wherein each Z is optionally substituted by OH, NH2, -CO2H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 2-6 Acyl, C 2-6 Alkanoic acid, C 2-6 Alkanoate or heterocyclic substitution, wherein the heterocyclic group, C 3-10 Cycloalkyl is a saturated or partially unsaturated 3-7 membered monocyclic, 6-10 membered bicyclic or 13-16 membered polycyclic ring system having 1, 2 or 3 heteroatom ring members independently selected from N, O and S, each optionally substituted with 1, 2, 3, 4 or 5 substituents each selected from R2;

[0034] R2 is independently selected from halogen, hydroxy, cyano, C 1-4 Alkyl, Hydroxyl-C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino, C 1-4 Alkylamino, (C 1- Alkyl)2amino, halo-C 1-4 Alkylamino, (halogenated-C 1-6 Alkyl)2amino, hydroxy-C 1-4 Alkylamino, C 1-4 Alkoxy-C 1-4 Alkyl-amino, amino-C 1-4 Alkyl, C 1-4 Alkylamino-C 1-4 Alkyl, (C 1-4 Alkylamino)2-C 1-4 Alkyl, C1-4 Alkoxy, halo-C 1-4 Alkoxy, Hydroxyl-C 1-4 Alkoxy, C 1-4 Alkyl-C 1-4 Alkoxy, C 3-10 Cycloalkyl, C 3-10 Cycloalkyl-amino, C 3-10 Cycloalkyl-amino-C 1-4 Alkyl, Heteroaryl-C 1-4 Alkyl, heteroarylamino, heteroaryl-C 1-4 Alkyl-amino, heterocyclyl, heterocyclyl-C 1-4 Alkyl, heterocyclyl-amino, heterocyclyl-amino-C 1-4 Alkyl, heterocyclic-C 1-4 Alkoxy, heterocyclyl-amino-C 3-10 Cycloalkyl, phenyl and phenyl-C 1-4 Alkoxy,

[0035] wherein heteroaryl is a 5-6 membered monocyclic or 6-10 membered bicyclic ring system having 1, 2, 3 or 4 heteroatom ring members independently selected from N, O or S,

[0036] wherein the heterocyclyl group is a saturated or partially unsaturated 3-7 membered monocyclic, 6-10 membered bicyclic or 13-16 membered polycyclic ring system having 1, 2 or 3 heteroatom ring members independently selected from N, O and S,

[0037] Among them C 3-10 Cycloalkyl is a saturated or partially unsaturated 3-7 membered monocyclic ring system, and

[0038] Each instance of phenyl, heteroaryl, heterocyclyl, or C 3-10 The cycloalkyl group is optionally substituted with 1 or 2 substituents each selected from R3;

[0039] R3 is independently selected from halogen, hydroxy, cyano, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino, C 1-4 Alkoxy and halo-C 1-4 Alkoxy;

[0040] Each R4 is independently selected from halogen, hydroxy, cyano, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino, C 1-4 Alkylamino, (C 1-4 Alkyl)2amino, C 1-4 Alkoxy, halo-C 1-4 Alkoxy, heteroaryl, heterocyclyl and phenyl,

[0041] wherein heteroaryl is a 5-6 membered monocyclic or 9-10 membered bicyclic ring system having 1, 2, 3 or 4 heteroatom ring members selected from N, O and S,

[0042] wherein heterocyclyl is a saturated or partially unsaturated 3-7 membered monocyclic or 9-10 membered bicyclic ring system having 1, 2 or 3 heteroatom ring members selected from N, O and S, and

[0043] wherein each instance of phenyl, heteroaryl or heterocyclyl is optionally substituted with 1 or 2 substituents each selected from R5;

[0044] R5 is independently selected from halogen, hydroxy, cyano, nitro, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino, C 1-4 Alkylamino, (C 1-4 alkyl)2amino, amino-C 1-4 Alkyl, Hydroxyl-C 1-4 Alkyl, C 1-4 Alkylcarbonyl, C 1-4 Alkoxy, C 1-4 Alkylthio, halo-C 1-4 Alkoxy and C 3-10 Cycloalkyl;

[0045] Wherein, the form of the compound is selected from the group consisting of pharmaceutically acceptable salts, hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers and tautomers thereof.

[0046] Another aspect of the present invention provides a compound having the structure of Formula II:

[0047]

[0048] in:

[0049] Each W is independently CH, CR' or N;

[0050] Each A is independently CH, CH2, CR a , CR4, N, NH or NR4;

[0051] Each -- represents a single or double bond;

[0052] Y is NR 1b or key;

[0053] Each R w For hydrogen, halogen, C 1-4 Alkyl, C 3-10 Cycloalkyl, C 3-10Cycloalkoxy, halo-C 1-4 Alkyl, C 1-4 Alkoxy or halogenated-C 1-4 Alkoxy,

[0054] Each R 1b are independently hydrogen, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl or hydroxy-C 1-4 alkyl;

[0055] Each Z is a heterocyclic group, a heteroaryl group, an aryl group, a C 3-10 Cycloalkyl, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, Hydroxyl-C 1-8 Alkyl, NH(hydroxy-C 1-6 Alkyl), NH(C 1-6 alkoxy), wherein each Z is optionally substituted by OH, NH2, -CO2H, halogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 2-6 Acyl, C 2-6 Alkanoic acid, C 2-6 Alkanoate or heterocyclyl substituted, wherein heterocyclyl is a saturated or partially unsaturated 3-7 membered monocyclic, 6-10 membered bicyclic or 13-16 membered polycyclic ring system having 1, 2 or 3 heteroatom ring members independently selected from N, O and S, each optionally substituted with 1, 2, 3, 4 or 5 substituents each selected from R2;

[0056] R2 is independently selected from halogen, hydroxy, cyano, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino, C 1-4 Alkylamino, (C 1- Alkyl)2amino, halo-C 1-4 Alkylamino, (halogenated-C 1-6 Alkyl)2amino, hydroxy-C 1-4 Alkylamino, C 1-4 Alkoxy-C 1-4 Alkyl-amino, amino-C 1-4 Alkyl, C 1-4 Alkylamino-C 1-4 Alkyl, (C 1-4 Alkylamino)2-C 1-4 Alkyl, C 1-4 Alkoxy, halo-C 1-4 Alkoxy, C 1-4Alkyl-C 1-4 Alkoxy, C 3-10 Cycloalkyl, C 3-10 Cycloalkyl-amino, C 3-10 Cycloalkyl-amino-C 1-4 Alkyl, Heteroaryl-C 1-4 Alkyl, heteroarylamino, heteroaryl-C 1-4 Alkyl-amino, heterocyclyl, heterocyclyl-C 1-4 Alkyl, heterocyclyl-amino, heterocyclyl-amino-C 1-4 Alkyl, heterocyclic-C 1-4 Alkoxy, heterocyclyl-amino-C 3-10 Cycloalkyl, phenyl and phenyl-C 1-4 Alkoxy,

[0057] wherein heteroaryl is a 5-6 membered monocyclic or 6-10 membered bicyclic ring system having 1, 2, 3 or 4 heteroatom ring members independently selected from N, O and S,

[0058] wherein the heterocyclyl group is a saturated or partially unsaturated 3-7 membered monocyclic, 6-10 membered bicyclic or 13-16 membered polycyclic ring system having 1, 2 or 3 heteroatom ring members independently selected from N, O and S,

[0059] Among them C 3-10 Cycloalkyl is a saturated or partially unsaturated 3-7 membered monocyclic ring system, and

[0060] Each instance of phenyl, heteroaryl, heterocyclyl, or C 3-10 The cycloalkyl group is optionally substituted with 1 or 2 substituents each selected from R3;

[0061] R3 is independently selected from halogen, hydroxy, cyano, C 1-4 Alkyl, deuterated-C 1-4 Alkyl, halo-C 1-4 Alkyl, amino, C 1-4 Alkoxy and halo-C 1-4 Alkoxy;

[0062] R' is selected from H, substituted or unsubstituted cycloalkyl, C 1-4 Alkyl, halo-C 1-4 Alkyl, halo-C 1-6 Alkoxy, halogen, C 1-6 Alkoxy, cyano, hydroxy-C 1-4 Alkyl, or aryl, optionally with C 1-4 Alkyl substitution;

[0063] Each R aand -C(=O)R2, -OC(=O)R2, -C(=O)OR2, -OC(=O)OR2, -C(=O)N(R2)2, -OC(=O)N(R2)2, -NR2C(=O)R2, -P(=O)(R2), C(=O)R2, -C(=O)R2, -C(=O)OR2, -OC(=O)OR2, -C(=O)N(R2)2, -OC(=O)N(R2)2, -NR2C(=O)R2, -P(=O)(R2), C(=O)R2, -C(=O)R2, -C(=O)OR2, -OC(=O)OR2, -C(=O)N(R2)2, -OC(=O)N(R2)2, - 1-4 Alkyl, (C 1-4 Alkyl)2, halo-C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-8 Cycloalkyl, C 2-7 heterocycloalkyl, aryl or monocyclic heteroaryl; and

[0064] R4 is independently selected from halogen, hydroxy, cyano or C 1-4 alkyl;

[0065] Wherein, the form of the compound is selected from the group consisting of pharmaceutically acceptable salts, hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers, tautomers, and isotopically enriched forms thereof.

[0066] The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound defined as a compound of formula I or a form thereof as disclosed herein, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers. The pharmaceutical composition is used for the treatment of diseases and / or conditions associated with NLRP3 activity. On the other hand, the present invention provides a composition, in particular a pharmaceutical composition, comprising a therapeutically effective amount of a compound defined as a compound of formula I or a subformula thereof as disclosed herein, or a pharmaceutically acceptable salt thereof, and one or more therapeutic agents. On the other hand, the present invention provides a composition, in particular a pharmaceutical composition as disclosed herein, for use as a medicament.

[0067] On the other hand, the present invention provides a compound of Formula I or its subformula as disclosed herein or a pharmaceutically acceptable salt thereof for treating a disease or condition, wherein the NLRP3 signal causes the pathology, and / or symptoms, and / or progression of the disease or condition. On the other hand, the present invention provides a method for treating a disease or condition, wherein the NLRP3 signal causes the pathology, and / or symptoms, and / or progression of the disease or condition, comprising administering a therapeutically effective amount of a compound of Formula I or its subformula as disclosed herein, or a pharmaceutically acceptable salt thereof. On the other hand, the present invention provides a method for inhibiting NLRP3 inflammasome activity in a subject in need thereof, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I or its form as disclosed herein or a pharmaceutically acceptable salt thereof.

[0068] Another aspect of the present invention relates to a compound of formula I or subformulae thereof as disclosed herein, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0069] Another aspect of the present invention relates to a compound of formula I or subformulae thereof as disclosed herein, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0070] Another aspect of the present invention also provides a compound of Formula I or its subformula as disclosed herein, or a pharmaceutically acceptable salt thereof, for use in treating a disease or condition selected from an inflammasome-related disease / condition, an immune disease, an inflammatory disease, an autoimmune disease, and an autoinflammatory disease. DETAILED DESCRIPTION

[0071] The present invention provides a compound selected from the group consisting of:

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087] Wherein, the form of the compound is selected from the group consisting of its pharmaceutically acceptable salt, hydrate, solvate, racemate, enantiomer, diastereomer, stereoisomer, tautomer, and isotope-enriched form. In embodiment 1, the present invention provides a compound as described above or a pharmaceutically acceptable salt thereof. In embodiment 2, the present invention provides a compound selected from the following:

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107] Wherein, the form of the compound is selected from the group consisting of pharmaceutically acceptable salts, hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers, tautomers, and isotopically enriched forms thereof.

[0108] In embodiment 3, the present invention provides a compound selected from the group consisting of:

[0109]

[0110]

[0111]

[0112] Therein, the form of the compound is selected from the group consisting of hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers, tautomers, free bases and isotopically enriched forms thereof.

[0113] In embodiment 4, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of embodiments 1 to 3 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0114] In embodiment 5, the present invention provides a method for treating or ameliorating a disease regulated by NLRP3 in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of embodiments 1 to 3.

[0115] In embodiment 6, the present invention provides a method for treating or ameliorating a disease regulated by NLRP3 according to embodiment 5, wherein the disease is selected from Alzheimer's disease, frontotemporal dementia (FTD), Huntington's disease, Parkinson's disease, perioperative neurocognitive disorder, cognitive disorder after cardiac arrest, cognitive disorder after stroke, sepsis, sepsis-related encephalopathy, subarachnoid hemorrhage, macular degeneration, retinal neovascularization, uveitis, colitis, endothelial dysfunction, gout, pseudogout, graft-versus-host disease (GvHD), systemic lupus erythematosus-lupus nephritis, cryopyrin-associated periodic syndrome (CAPS), cystic fibrosis, sickle cell disease, VCP-related diseases, liver fibrosis, non-alcoholic fatty liver disease (NASH), muscle atrophy, inherited and acquired myopathies such as Duchenne muscular dystrophy (DMD), hyperalgesia, neuropathic pain in multiple sclerosis, acute kidney injury, chronic crystalline nephropathy, chronic kidney disease, asthma and allergic airway inflammation, diabetes-related atherosclerosis, diabetic encephalopathy, diabetic nephropathy, islet transplant rejection, obesity-related nephropathy, oxalate nephropathy, renal fibrosis, renal hypertension, type I diabetes, type II diabetes, psoriasis, hidradenitis suppurativa, atherosclerosis, and cytokine release syndrome (CRS).

[0116] In embodiment 7, the invention provides the method of any one of embodiments 5 to 6, wherein the effective amount of the compound ranges from about 0.001 mg / kg / day to about 500 mg / kg / day.

[0117] In embodiment 8, the present invention provides a compound according to any one of embodiments 1 to 3 or a pharmaceutically acceptable salt thereof for use in treating or ameliorating a disease regulated by NLRP3, wherein the disease is selected from Alzheimer's disease, frontotemporal dementia (FTD), Huntington's disease, Parkinson's disease, perioperative neurocognitive disorder, cognitive disorder after cardiac arrest, cognitive disorder after stroke, sepsis, sepsis-related encephalopathy, subarachnoid hemorrhage, macular degeneration, retinal neovascularization, uveitis, colitis, endothelial dysfunction, gout, pseudogout, graft-versus-host disease (GvHD), systemic lupus erythematosus-lupus nephritis, and cold pyrine-related periodic syndrome. syndrome (CAPS), cystic fibrosis, sickle cell disease, VCP-related diseases, liver fibrosis, nonalcoholic fatty liver disease (NASH), muscular dystrophy, inherited and acquired myopathies, hyperalgesia, neuropathic pain in multiple sclerosis, acute kidney injury, chronic crystalline nephropathy, chronic kidney disease, asthma and allergic airway inflammation, diabetes-related atherosclerosis, diabetic encephalopathy, diabetic nephropathy, islet transplant rejection, obesity-related nephropathy, oxalate nephropathy, renal fibrosis, renal hypertension, type I diabetes, type II diabetes, psoriasis, hidradenitis suppurativa, atherosclerosis, and cytokine release syndrome (CRS).

[0118] In embodiment 9, the invention provides the use of a compound according to embodiment 8, wherein the effective amount of the compound ranges from about 0.001 mg / kg / day to about 500 mg / kg / day.

[0119] In embodiment 10, the present invention provides the use of a compound according to any one of embodiments 1 to 3 in the preparation of a pharmaceutical composition for treating or ameliorating a disease regulated by NLRP3 in a subject in need thereof, comprising administering to the subject an effective amount of the compound or a form thereof mixed with one or more pharmaceutically acceptable excipients.

[0120] Methods of using the present invention

[0121] There is evidence that NLRP3-induced IL-1β and IL-18 play a role in inflammatory responses associated with or caused by a variety of different diseases (Menu et al., Clinical and Experimental Immunology, 2011, 166, 1-15; Strowig et al., Nature, 2012, 481, 278-286). NLRP3 mutations have been found to cause a series of rare autoinflammatory diseases CAPS (Ozaki et al., J. Inflammation Research, 2015, 8, 15-27; Schroder et al., Cell, 2010, 140: 821-832; Menu et al., Clinical and Experimental Immunology, 2011, 166, 1-15). CAPS is a genetic disease characterized by recurrent fever and inflammation, consisting of three autoinflammatory diseases that form a clinical continuum. These diseases, in order of increasing severity, are familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), and chronic infantile neurocutaneous arthritis (CINCA; also known as neonatal onset multisystem inflammatory disease, NOMID), all of which have been shown to be caused by gain-of-function mutations in the NLRP3 gene, which result in increased secretion of IL-1β. NLRP3 has also been implicated in a variety of autoinflammatory diseases, including suppurative arthritis, pyoderma gangrenosum, and acne (PAPA), Sweet syndrome, chronic nonbacterial osteomyelitis (CNO), and acne vulgaris (Cook et al., Eur J. Immunol., 2010, 40, 595-653). A variety of autoimmune diseases have been shown to be associated with NLRP3, including, in particular, multiple sclerosis, type 1 diabetes (T1D), psoriasis, rheumatoid arthritis (RA), Behcet's syndrome, Schnitzler syndrome, macrophage activation syndrome (Braddock et al., Nat. Rev. Drug Disc. 2004, 3, 1-10; Inoue et al., Immunology, 2013, 139, 11-18, Coll et al., Nat. Med. 2015, 21(3), 248-55; Scott et al., Clin. Exp. Rheumatol. 2016, 34(1), 88-93), systemic lupus erythematosus and its complications, such as lupus nephritis (Lu et al., J. Immunol., 2017, 198(3), 1119-29) and systemic sclerosis (Artlett et al., Arthritis Rheum.2011,63(11),3563-74).NLRP3 has also been shown to play a role in a variety of lung diseases, including chronic obstructive pulmonary disease (COPD), asthma (including steroid-resistant asthma), asbestosis, and silicosis (De Nardo et al., Am. J. Pathol., 2014, 184:42-54; Kim et al., Am. J. Respir Crit Care Med, 2017, 196(3), 283-97). NLRP3 is also thought to play a role in a variety of central nervous system diseases, including multiple sclerosis (MS), Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, brain damage caused by pneumococcal meningitis (Walsh et al., Nature Reviews, 2014, 15, 84-97; and Dempsey et al., Brain. Behav. Immun. 2017, 61, 306-16), intracranial aneurysms (Zhang et al., J. Stroke and Cerebrovascular Dis., 2015, 24, 5, 972-9), and traumatic brain injury (Ismael et al., J. Neurotrauma., 2018, 35(11), 1294-1303). NRLP3 activity has also been shown to be associated with a variety of metabolic diseases, including type 2 diabetes (T2D) and its organ-specific complications, atherosclerosis, obesity, gout, pseudogout, metabolic syndrome (Wen et al., Nature Immunology, 2012, 13, 352-357; Duewell et al., Nature, 2010, 464, 1357-1361; Strowig et al., Nature, 2014, 481, 278-286), and non-alcoholic fatty liver disease (Mridha et al., J. Hepatol. 2017, 66 (5), 1037-46). NLRP3 is also believed to play a key pathological role in the occurrence and development of a variety of skeletal muscle diseases, such as muscle atrophy, hereditary and acquired myopathy (Dubussion et al., Cells 2021, 10 (11): 3023). NLRP3 is also thought to play a role in atherosclerosis, myocardial infarction (van Hout et al., Eur Heart J. 2017, 38(11), 828-3-6), heart failure (Sano et al., J. Am. Coll. Cardiol. 2018, 71(8), 875-66), aortic aneurysm and dissection (Wu et al., Arterioscler Thromb. Vase. Biol., 2017, 37(4), 694-706), and other cardiovascular events (Ridker et al., N. Engl. J. Med, 2017, 377(12), 1119-31) through IL-1β.Other diseases that have been shown to be associated with NLRP3 include: ocular diseases such as wet and dry age-related macular degeneration (Doyle et al., Nature Medicine, 2012, 18, 791-798; Tarallo et al., Cell 2012, 149(4), 847-59), diabetic retinopathy (Loukovaara et al., Acta Ophthalmol., 2017, 95(8), 803-8), non-infectious uveitis and optic nerve damage (Puyang et al., Sci. Rep. 2016, 6, 20998); liver diseases, including non-alcoholic steatohepatitis (NASH) and acute alcoholic hepatitis (Henao-Meija et al., Nature, 2012, 482, 179-185); inflammatory reactions in the lungs and skin (Primiano et al., J. Immunol. 2016, 197(6), 2421-33), including contact allergies (such as bullous pemphigoid (Fang et al., JDermatol Sci. 2016, 83(2), 116-23)), atopic dermatitis (Niebuhr et al., Allergy, 2014, 69(8), 1058-67), suppurative hidradenitis (Alikhan et al., J. Am. Acad. Dermatol., 2009, 60(4), 539-61), and sarcoidosis (Jager et al., Am. J. Respir Crit. Care Med., 2015, 191, A5816); joint inflammatory reactions (Braddock et al., Nat. Rev. Drug Disc, 2004, 3, 1-10); amyotrophic lateral sclerosis (Gugliandolo et al., Int. J. Mol. Sci., 2018, 19(7), E1992); cystic fibrosis (larmitti et al., Nat. Commun., 2016, 7, 10791); stroke (Walsh et al., Nature Reviews, 2014, 15, 84-97); chronic kidney disease (Granata et al., PLoS One 2015, 10(3), eoi22272); inflammatory bowel disease, including ulcerative colitis and Crohn's disease (Braddock et al., Nat. Rev. Drug Disc, 2004, 3, 1-10; Neudecker et al., J. Exp. Med. 2017, 214(6), 1737-52; Lazaridis et al., Dig. Dis. Sci. 2017, 62(9), 2348-56). NLRP3 inflammasome was found to be activated in response to oxidative stress.NLRP3 has also been shown to be associated with inflammatory hyperalgesia (Dolunay et al., Inflammation, 2017, 40, 3-66-86). U.S. application US202003-61898 is incorporated herein by reference.

[0122] definition

[0123] To help understand the scope of the compounds of Formula I described herein, or forms thereof, the following specific examples are included. Of course, experiments related to the compounds of Formula I described herein, or forms thereof, should not be interpreted as specifically limiting the scope of the compounds of Formula I described herein, or forms thereof, and such variations of the compounds of Formula I described herein, or forms thereof, whether now known or later developed, as long as they are within the grasp of those skilled in the art, are considered to fall within the scope of the claims described herein and below.

[0124] Except working examples, unless otherwise stated, all numerals representing the quantity of components, reaction conditions, experimental data, etc. used in the specification and claims should be understood to be modified by the term "about". Thus, all these numerals represent approximate values, which can vary according to the expected characteristics to be obtained by the reaction or due to the experimental conditions of the change. Therefore, within the expected range of experimental reproducibility, the term "about" refers to the data range provided in the context of the result data, which can vary according to the standard deviation of the mean value. Similarly, for the experimental results provided, the obtained data can be rounded, so as to present the data consistently without losing significant figures. At least it is not intended to limit the application of the principle of equivalents within the scope of the claims, and each numerical parameter should be interpreted according to the number of significant figures and conventional rounding algorithms.

[0125] Although the numerical ranges and parameters describing the characteristics of the compounds of Formula I or forms thereof described herein are approximations, the numerical values ​​described in the working examples are as precise as possible. However, any numerical value inherently contains certain errors caused by the standard deviation found in its respective testing measurements.

[0126] The compounds of formula I provided herein or their forms will be described in more detail with reference to the following non-limiting examples, which are provided to more fully illustrate the scope of the compounds of formula I described herein or their forms, but are not to be construed as limiting their scope. The examples illustrate the preparation of the compounds of formula I described herein or their forms, and the testing of these compounds of formula I or their forms in vitro and / or in vivo. It will be appreciated by those skilled in the art that the synthetic techniques described in these examples represent techniques within the practical scope of ordinary technicians in the field of chemistry, and therefore constitute the preferred mode of their practice. However, it should be understood that, according to the present invention, those with the skills in the art should recognize that many changes can be made in the specific methods disclosed herein without departing from the subject matter and scope described herein, while still obtaining similar or similar results.

[0127] In certain embodiments described herein, the compound of Formula I or a form thereof is isolated for use.

[0128] As used herein, the term "isolated" refers to the physical state of a compound of Formula I or a form thereof after separation and / or separation and / or purification from a synthetic process (e.g., from a reaction mixture) or a natural source or a combination thereof, according to a separation, separation or purification process or method described herein or known to a skilled person (e.g., chromatography, recrystallization, etc.), to a purity sufficient to be characterized by standard analytical techniques described herein or known to a skilled person.

[0129] As used herein, the term "protected" refers to a compound of formula I or a functional group thereof that is in a modified form when the compound is reacted to exclude undesired side reactions of the functional group. Suitable protective groups will be identified by one of ordinary skill in the art, as well as by reference to standard textbooks, such as TW Greene et al., Protective Groups in Organic Synthesis (2007), Wiley, New York.

[0130] Also contemplated are prodrugs and solvates of the compounds of Formula I described herein, or forms thereof.

[0131] As used herein, the term "prodrug" refers to a compound of formula I or a functional group thereof in a form that is in a state of being transformed in vivo to produce an active or more active compound of formula I or a form thereof (e.g., as an active or inactive drug precursor). The transformation can occur by various mechanisms (e.g., by metabolic and / or non-metabolic chemical processes), for example, by hydrolysis and / or metabolism in blood, liver and / or other organs and tissues. Discussion on the use of prodrugs is provided by VJ Stella et al., "Biotechnology: Pharmaceutical Aspects, Prodrugs: Challenges and Rewards," American Association of Pharmaceutical Scientists and Springer Press, 2007.

[0132] In one embodiment, when the compound of Formula I or its form contains a carboxylic acid functional group, the prodrug may include an ester formed by replacing the hydrogen atom of the acid group with a functional group such as an alkyl group. In another embodiment, when the compound of Formula I or its form contains an alcohol functional group, the hydrogen atom of the alcohol group may be replaced with a functional group such as an alkyl or carbonyl group to form a prodrug. In another embodiment, when the compound of Formula I or its form contains an amine functional group, one or more amine hydrogen atoms may be replaced with a functional group such as an alkyl or substituted carbonyl group to form a prodrug.

[0133] Pharmaceutically acceptable prodrugs of a compound of formula I or a form thereof include those substituted with one or more of the following groups: carboxylate, sulfonate, amino acid ester, phosphonate (e.g., ammonium metaphosphate for the derivatization of ammonium metaphosphate) and monophosphate, diphosphate or triphosphate further substituted with an alkyl group (at an appropriate position). As described herein, it will be appreciated by those of ordinary skill in the art that one or more such substituents may be used to provide a compound of formula I or a form thereof as a prodrug.

[0134] The compound of formula I or its form can form salts, which are intended to be included within the scope of this specification. Unless otherwise indicated, reference to the compound of formula I or its form is understood to include reference to its salt. The term "salt" as used herein refers to an acidic salt formed with an inorganic acid and / or an organic acid, and a basic salt formed with an inorganic base and / or an organic base. In addition, when the compound of formula I or its form contains a basic group (such as, but not limited to, pyridine or imidazole) and an acidic group (such as, but not limited to, a carboxylic acid) at the same time, a zwitterion ("inner salt") can be formed and is included in the term "salt" as used herein.

[0135] As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of Formula I or a form thereof that is safe, effective (i.e., non-toxic, physiologically acceptable), and biologically active for use in mammals, although other salts are also useful. Salts of a compound of Formula I or a form thereof can be formed, for example, by reacting a compound of Formula I or a form thereof and a compound described herein with an amount (e.g., an equal amount) of an acid or base in a medium (such as a medium in which the salt is precipitated or in an aqueous medium), followed by lyophilization.

[0136] In addition, acids that are considered suitable for forming pharmaceutically useful salts of basic drug compounds are discussed, for example, in P. Stahl et al., Camille G. (eds.) Handbook of Pharmaceutical Salts. Properties, Selection and Use. (2002) Zurich: Wiley-VCH; S. Berge et al., Journal of Pharmaceutical Sciences (1977) 66 (1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33, 201-217; Anderson et al., The Practice of Medicinal Chemistry (1996), Academic Press, New York; and in The Orange Book (Food and Drug Administration website, Washington, D.C.). These disclosures are incorporated herein by reference.

[0137] Suitable alkaline salts include, but are not limited to, aluminum, ammonium, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts. Certain compounds of Formula I described herein or forms thereof may also be quaternized with organic bases (e.g., organic amines), such as, but not limited to, dicyclohexylamine, tert-butylamine, and the like, and with various amino acids, such as, but not limited to, arginine, lysine, and the like. Basic nitrogen-containing groups may be quaternized using, for example, lower alkyl halides (e.g., methyl, ethyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl, diethyl, and dibutyl sulfates), long chain halides (e.g., decyl, dodecyl, and stearyl chlorides, bromides, and iodides), aralkyl halides (e.g., benzyl and phenethyl bromides), and other agents.

[0138] All such acid salts and base salts are intended to be encompassed within the scope of the pharmaceutically acceptable salts described herein. In addition, for the purposes of this specification, all such acid salts and base salts are considered equivalent to the free forms of the corresponding compounds.

[0139] The compounds of Formula I and forms thereof may further exist as tautomers. All such tautomeric forms are contemplated and intended to be included within the scope of the compounds of Formula I or forms thereof as described herein.

[0140] The compounds of formula I or forms thereof may contain asymmetric or chiral centers and may therefore exist in different stereoisomeric forms. The present specification is intended to include all stereoisomeric forms of the compounds of formula I or forms thereof, as well as mixtures thereof, including racemic mixtures.

[0141] The compounds of Formula I described herein or forms thereof may include one or more chiral centers and thus may exist as racemic mixtures (R / S) or substantially pure enantiomers and diastereomers. The compounds may also exist as substantially pure (R) or (S) enantiomers (when there is one chiral center). In one embodiment, the compounds of Formula I described herein or forms thereof are (S) isomers and may exist as enantiomerically pure compositions that substantially contain only (S) isomers. In another embodiment, the compounds of Formula I described herein or forms thereof are (R) isomers and may exist as enantiomerically pure compositions that substantially contain only (R) isomers. As one skilled in the art will recognize, when there are more than one chiral center, the compounds of Formula I described herein or forms thereof may also exist as (R,R), (R,S), (S,R) or (S,S) isomers as defined by the IUPAC nomenclature recommendations.

[0142] As used herein, the term "substantially pure" means that the compound of Formula I or a form thereof consists essentially of a single isomer, wherein the content of the single isomer is greater than or equal to 90%, the content is greater than or equal to 92%, the content is greater than or equal to 95%, the content is greater than or equal to 98%, the content is greater than or equal to 99%, or the content is equal to 100%.

[0143] In one aspect of the specification, the compound of Formula I or a form thereof is a substantially pure (S) enantiomer, present at a content of greater than or equal to 90%, a content of greater than or equal to 92%, a content of greater than or equal to 95%, a content of greater than or equal to 98%, a content of greater than or equal to 99%, or a content of equal to 100%.

[0144] In one aspect of the specification, the compound of Formula I or a form thereof is a substantially pure (R) enantiomer, present at a content greater than or equal to 90%, a content greater than or equal to 92%, a content greater than or equal to 95%, a content greater than or equal to 98%, a content greater than or equal to 99%, or a content equal to 100%.

[0145] As used herein, the term "racemate" refers to any mixture of isomeric forms that are not "enantiomerically pure", including, for example, but not limited to, mixtures in ratios of about 50 / 50, about 60 / 40, about 70 / 30, or about 80 / 20, about 85 / 15, or about 90 / 10.

[0146] In addition, the compounds of Formula I described herein or forms thereof include all geometric and positional isomers. For example, if the compounds of Formula I or forms thereof contain double bonds or fused rings, the cis- and trans-forms and mixtures thereof are included within the scope of the compounds of Formula I described herein or forms thereof.

[0147] Diastereomeric mixtures can be separated into their individual diastereomers based on their physical chemical differences by methods well known to those skilled in the art, for example, by chromatography and / or fractional crystallization. Enantiomers can be separated by using chiral high performance liquid chromatography columns or other chromatographic methods known to those skilled in the art.

[0148] Enantiomers may also be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers.

[0149] All stereoisomers (e.g., geometric isomers, optical isomers, etc.) of existing compounds of Formula I or its forms (including salts, solvates, esters and prodrugs and their converted prodrugs) that may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotational isomers, atropisomers, diastereoisomers and positional isomers, are considered within the scope of the present specification. For example, a single stereoisomer of a compound of Formula I or its form described herein may be substantially free of other isomers, or may be present in a racemic mixture as described above.

[0150] The use of the terms "salt," "solvate," "ester," "prodrug," and the like are intended to apply equally to the enantiomers, stereoisomers, rotational isomers, tautomers, positional isomers, racemates, isotopically enriched or prodrug salts, solvates, esters and prodrugs of the present compounds.

[0151] One or more compounds of Formula I or forms thereof described herein may exist in unsolvated forms as well as solvated forms with pharmaceutically acceptable solvents (eg, water, ethanol, etc.), and the description herein is intended to encompass both solvated and unsolvated forms.

[0152] As used herein, the term "solvate" refers to a physical association of a compound of Formula I described herein, or a form thereof, with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, the solvate will be able to separate, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. As used herein, "solvate" includes solution phase solvates and separable solvates. Non-limiting examples of suitable solvates include glycolates, methanolates, and the like.

[0153] One or more compounds of Formula I described herein or forms thereof may be optionally converted into solvates. The preparation of solvates is well known. A typical non-limiting method includes dissolving a compound of Formula I or a form thereof in a desired amount of a desired solvent (organic matter or water or a mixture thereof) at a temperature above ambient temperature, and cooling the solution at a rate sufficient to form crystals, and then separating the crystals by standard methods. Analytical techniques (e.g., infrared spectroscopy) show that the solvent (or water) is present as a solvate (or hydrate) in the crystal.

[0154] As used herein, the term "hydrate" refers to a solvate wherein the solvent molecule is water.

[0155] Polymorphs and amorphous forms of the compounds of Formula I or forms thereof, as well as salts, solvates, esters, and prodrugs of the compounds of Formula I or forms thereof, are further intended to be included within the scope of the compounds of Formula I or forms thereof described herein.

[0156] As used herein, the term "isotopically enriched" refers to a compound of Formula I or a form thereof that is identical to a compound of Formula I or a form thereof described herein, but in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into a compound of Formula I or a form thereof described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as H, 2 , H 3 , C 13 , C 14 、N 15 , O 18 , O 17 , P 31 , P 32 , S 35 、F 18 , Cl 35 and Cl 36 , each of which is also within the scope of this specification.

[0157] Dosage and administration

[0158] The compounds of the present invention can be formulated into a variety of oral dosage forms and carriers. Oral administration can be in the form of tablets, coated tablets, dragées, hard and soft gelatin capsules, solutions, emulsions, syrups or suspensions. The compounds of the present invention are effective when administered by other routes of administration (including continuous (intravenous infusion) topical parenteral, intramuscular, intravenous, subcutaneous, transdermal (which may include penetration enhancers), oral, nasal, inhalation and suppositories, etc.). The preferred mode of administration is usually oral, with a convenient daily dosing regimen that can be adjusted according to the degree of the disease and the patient's response to the active ingredient.

[0159] One or more compounds of the present invention and pharmaceutically acceptable salts thereof, together with one or more conventional excipients, carriers or diluents, can be prepared into the form of pharmaceutical compositions and unit doses. Pharmaceutical compositions and unit dosage forms can be composed of conventional ingredients in conventional proportions, with or without other active compounds or principles, and unit dosage forms can contain any suitable effective amount of active ingredients commensurate with the intended daily dose range to be used. Pharmaceutical compositions can be used as solids (e.g., tablets or filled capsules, semisolids, powders, sustained-release preparations) or liquids (e.g., solutions, suspensions, emulsions, elixirs or filled capsules for oral use); or in the form of suppositories for rectal or vaginal administration; or in the form of sterile injection solutions for parenteral use. Typical preparations will contain about 5% to about 95% of one or more active compounds (w / w). The term "preparation" or "dosage form" is intended to include solid and liquid preparations of active compounds, and those skilled in the art will understand that the active ingredient can be present in different preparations depending on the target organ or tissue and the desired dose and pharmacokinetic parameters.

[0160] The term "excipient" as used herein refers to compounds that are generally safe, non-toxic, and have no adverse effects biologically or otherwise, and include excipients acceptable for veterinary and human pharmaceutical uses. The compounds of the present invention can be administered alone, but will usually be administered in admixture with one or more suitable pharmaceutical excipients, diluents or carriers selected according to the intended route of administration and standard pharmaceutical practice.

[0161] "Pharmaceutically acceptable" means useful for preparing a pharmaceutical composition that is generally safe, non-toxic, and has no biologically or otherwise adverse effects, and includes pharmaceutical compositions that are acceptable for veterinary and human pharmaceutical use.

[0162] The "pharmaceutically acceptable salt" form of the active ingredient may also initially impart desirable pharmacokinetic properties to the active ingredient that the non-salt form does not possess, and may even positively affect the pharmacodynamics of the active ingredient in terms of its in vivo therapeutic activity. The phrase "pharmaceutically acceptable salt" of a compound refers to a salt that is pharmaceutically acceptable and possesses the desired pharmacological activity of the parent compound. Such salts include: (1) salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.); or with organic acids, such as acetic acid, propionic acid, hexanoic acid, cyprolic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid (1) an acid addition salt formed when the acidic proton present in the parent compound is replaced by a metal ion, for example, an alkali metal ion, an alkaline earth ion or an aluminum ion, or coordinated with an organic base, for example, ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, etc.

[0163] Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories and dispersible granules. Solid carriers can be one or more substances, which can also serve as diluents, flavoring agents, solubilizers, lubricants, suspending agents, adhesives, preservatives, tablet disintegrants or encapsulating materials. In powders, the carrier is usually a subdivided solid, which is a mixture with the subdivided active ingredient. In tablets, the active ingredient is usually mixed with a carrier with the necessary binding capacity in an appropriate proportion and compressed into a desired shape and size. Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting point wax, cocoa butter, etc. In addition to the active ingredient, solid form preparations can also contain colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, etc.

[0164] Liquid preparations suitable for oral administration also include liquid preparations of emulsions, syrups, elixirs, aqueous solutions, aqueous suspensions. These liquid preparations include solid form preparations intended to be converted into liquid form preparations shortly before use. Emulsions can be prepared in solution, for example in propylene glycol aqueous solution, or can contain emulsifiers, for example lecithin, sorbitol monooleate or acacia. The aqueous solution can be prepared by dissolving the active ingredient in water, and adding suitable coloring agents, spices, stabilizers and thickening agents. Aqueous suspensions can be prepared by dispersing the subdivided active ingredient in water containing viscous materials (for example natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose and other well-known suspending agents).

[0165] The compounds of the present invention can be formulated for parenteral administration (e.g., by injection, such as bolus injection or continuous infusion), and can be present in a unit dose form in an ampoule, a prefilled syringe, a small volume infusion set, or a multi-dose container containing an added preservative. The composition can take the form of a suspension, solution, or emulsion in an oily or aqueous vehicle, such as a solution in a polyethylene glycol aqueous solution. Examples of oily or non-aqueous vehicles, diluents, solvents, or carriers include propylene glycol, polyethylene glycol, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate), and can contain formulation agents such as preservatives, wetting agents, emulsifiers or suspending agents, stabilizers, and / or dispersants. In addition, the active ingredient can be in powder form, obtained by aseptically separating sterile solids or by lyophilizing from a solution, and combined with a suitable carrier (e.g., sterile, pyrogen-free water) before use.

[0166] The compound of the present invention can be formulated into an ointment, cream or emulsion for topical administration to the epidermis, or as a transdermal patch. For example, ointments and emulsions can be prepared with an aqueous or oily base, and suitable thickeners and / or gelling agents are added. Emulsions can be prepared with an aqueous or oily base, and usually also contain one or more emulsifiers, stabilizers, dispersants, suspending agents, thickeners or coloring agents. Preparations suitable for topical administration in the oral cavity include: lozenges of an active agent contained in a flavoring matrix (usually sucrose and acacia or tragacanth), lozenges of an active ingredient contained in an inert matrix (e.g., gelatin and glycerol or sucrose and acacia), and mouthwashes of an active ingredient contained in a suitable liquid carrier.

[0167] The compounds of the present invention can be formulated into suppositories for administration. First, a low melting point wax (e.g., a mixture of fatty acid glycerides or cocoa butter) is melted, and the active ingredient is evenly dispersed by, for example, stirring. The molten homogeneous mixture is then poured into a mold of suitable size, cooled and solidified.

[0168] The compounds of the present invention may be formulated for vaginal administration as pessaries, tampons, creams, gels, pastes, foams or sprays containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

[0169] The compounds of the present invention can be formulated for nasal administration. The solution or suspension is directly applied to the nasal cavity by conventional methods (e.g., using a dropper, a pipette or a sprayer). The preparation can be provided in a single dose or multiple dose form. In the latter case of a dropper or a pipette, this can be achieved by the patient applying a suitable, predetermined volume of solution or suspension. In the case of a sprayer, this can be achieved by, for example, a metered atomizing spray pump.

[0170] The compounds of the present invention may be formulated for aerosol administration, particularly for the respiratory tract and including intranasal administration. The compounds typically have a small particle size, for example of the order of five (5) microns or less. Such particle sizes may be obtained by methods known in the art, for example by micronization. The active ingredient is provided in a pressurized package with a suitable propellant (e.g., a chlorofluorocarbon (CFC), such as dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, or carbon dioxide or other suitable gas). The aerosol may also conveniently contain a surfactant, such as lecithin. The dose of the drug may be controlled by a metering valve. Alternatively, the active ingredient may be provided in the form of a dry powder, such as a powder mixture of the compound in a suitable powder matrix (e.g., lactose, starch, starch derivatives, such as hydroxypropylmethylcellulose and polyvinylpyrrolidone (PVP)). The powder carrier forms a gel in the nasal cavity. The powder composition may be presented in a unit dose form, for example, in a capsule, or a cartridge in the form of a gelatin or blister pack, from which the powder may be administered by means of an inhaler.

[0171] If necessary, the preparation can be prepared with an enteric coating for sustained release or controlled release administration of active ingredients. For example, the compound of the present invention can be formulated in a transdermal or subcutaneous drug delivery device. When the sustained release of the compound is required and when the patient's compliance with the treatment regimen is of vital importance, these drug delivery systems are advantageous. The compound in the transdermal drug delivery system is usually attached to a skin adhesive solid support. The target compound can also be combined with a penetration enhancer, such as azone (1-dodecylazacycloheptane-2-ketone). The sustained release drug delivery system is inserted subcutaneously to the dermis by surgery or injection. The subcutaneous implant is wrapped in a fat-soluble film (for example, silicone rubber) or a biodegradable polymer (for example, polylactic acid).

[0172] Suitable formulations are described in Remington: The Science and Practice of Pharmacy 1995, edited by EW Martin, Mack Publishing Company, 19th edition, Easton, Pennsylvania, together with pharmaceutical carriers, diluents and excipients. Skilled formulation scientists can modify the formulation within the teachings of the specification to provide a variety of formulations for specific routes of administration without making the compositions of the invention unstable or compromising their therapeutic activity.

[0173] Modification of the present compound to make it more soluble in water or other carriers, for example, can be easily achieved by minor modifications (salt preparation, esterification, etc.), which are all within the ordinary skill of the art. It is also within the ordinary skill of the art to change the route and dosage regimen of a particular compound in order to manage the pharmacokinetics of the compound so that it produces the greatest beneficial effect on the patient.

[0174] The term "therapeutically effective amount" as used herein refers to the amount required to alleviate the symptoms of an individual disease. The dosage will be adjusted according to the individual needs of each specific case. Depending on a variety of factors, such as the severity of the disease to be treated, the age and general health of the patient, other drugs that the patient is being treated for, the route of administration and form, and the preferences and experience of the relevant doctors, this dosage can vary within a wide range of limitations. For oral administration, in monotherapy and / or combination therapy, a daily dose of about 0.01 mg / kg to about 1000 mg / kg body weight per day is appropriate. The preferred daily dose is between about 0.1 mg / kg and about 500 mg / kg body weight, more preferably, between 0.1 mg / kg and about 100 mg / kg body weight, and most preferably, between 1.0 mg / kg and about 10 mg / kg body weight. Therefore, for the administration of a 70 kg person, the dosage range is about 7 mg to 0.7 g per day. The daily dose can be administered as a single dose or divided doses, typically between 1 and 5 doses per day. Typically, the dose at the beginning of treatment is small, which is less than the optimal dose of the compound. Thereafter, the dosage is increased in small increments until the optimum therapeutic effect for the individual patient is reached. One of ordinary skill in treating the diseases described herein will be able to determine the therapeutically effective amount of the compounds of the invention for a given disease and patient without undue experimentation and relying on personal knowledge, experience, and the disclosure of this application.

[0175] The pharmaceutical formulation is preferably in unit dosage form. In this form, the formulation is subdivided into unit doses containing appropriate amounts of the active ingredient. The unit dosage form can be a packaged formulation containing discrete quantities of the formulation, such as packaged tablets, capsules, and powders in vials or ampoules. Alternatively, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or an appropriate number of any of these packaged forms.

[0176] The examples of representative compounds included in the present invention and within the scope of the present invention are provided in the following table. The following examples and preparations are provided to enable those skilled in the art to more clearly understand and practice the present invention. They should not be considered as limiting the scope of the present invention, but only as its description and representation.

[0177] In general, the nomenclature used in the present invention is based on AUTONOMTM v.4.0, a computer system used by the Beilstein Institute to generate the IUPAC systematic nomenclature. If there is a discrepancy between a depicted structure and the name given to the structure, the depicted structure is given more weight. In addition, if the stereochemistry of a structure or a portion of a structure is not indicated, for example, by bold or dashed lines, the structure or portion of a structure should be interpreted as comprising all of its stereoisomers.

[0178] Unless otherwise noted, reagents and solvents were used as purchased (from various suppliers). Where appropriate, as in the following examples, the term "Celite" is used to represent the trade name (Diatomaceous earth brand). Where appropriate, chromatographic separations were performed using commonly used techniques and equipment, such as using an ISCO Where appropriate, NMR spectra were obtained using conventional techniques and equipment, for example by using a Bruker Avance III 500 Spectrometer, using deuterated solvents (e.g. DMSO-d6 or residual solvents) as standards. Where appropriate, melting points were determined using commonly used techniques and equipment, e.g. using SRS MPA100 (values ​​obtained without correction / calibration). Where appropriate, TLC analysis was performed using commonly used techniques and equipment, such as by using Aldrich 254 nm glass backing plates ( 250 μm) and visualized using UV and I2 staining. Where appropriate, ESI mass spectra were acquired using commonly used techniques and equipment, such as using ACQUITY System, unless otherwise stated, values ​​are shown as [M+H] + or [MH] - Where appropriate, the structures of the products were obtained by 2D NOESY (Nuclear Overhauser Effect Spectroscopy) experiments.

[0179] Compounds and preparation

[0180] The examples of representative compounds included in the present invention and within the scope of the present invention are provided herein. The following examples and preparations are provided to enable those skilled in the art to more clearly understand and practice the present invention. They should not be considered to limit the scope of the present invention, but only as its description and representation.

[0181] experiment

[0182] As disclosed herein, the general method for preparing the compounds of Formula I described herein or forms thereof can be prepared using the methods summarized in Schemes AJ by appropriately selecting reagents, solvents, temperatures, pressures, and other reaction conditions that are readily selected by those of ordinary skill in the art with appropriate substitutions. Many starting materials are commercially available, or when not available, can be prepared by standard, well-known synthetic methods or using techniques known to those skilled in the art using the approaches described below. The synthetic schemes provided herein include multiple reaction steps, each of which is intended to be performed independently and can be performed with or without any prior or subsequent steps. In other words, each individual reaction step of the synthetic scheme provided separately herein is considered.

[0183] Depending on the nature of the groups described in the following schemes, the final compound or its precursor can be further described using standard, well-known synthetic methods (e.g., SN2 displacement reactions, metal-catalyzed coupling reactions (e.g., Suzuki coupling, Negishi coupling, and Buchwald coupling), reductive amination reactions, etc.) to obtain compounds of general formula I-II.

[0184] Option A:

[0185]

[0186] Compound A1 (wherein X1 and X2 are independently bromine, chlorine, etc.) is converted into compound A2 by nucleophilic substitution with a suitable amine (in the presence of a suitable base (such as DIEPA, etc.) in a suitable solvent (such as NMP, etc.)) or with a suitable alcohol (in the presence of a suitable base (such as NaH, etc.) in a suitable solvent (such as anhydrous THF, etc.). In the presence of a catalyst (such as Pd(dppf)Cl2, etc.) and a base (such as K2CO3 aqueous solution, etc.), in a suitable solvent (such as 1,4-dioxane, etc.), compound A2 is converted into compound A3 by Suzuki coupling with an aryl or heteroaryl boronic acid (or boronic acid pinacol ester).

[0187] Option B:

[0188]

[0189] In the presence of a catalyst (e.g., Pd(dppf)Cl2, etc.) and a base (e.g., K2CO3 aqueous solution, etc.), in a suitable solvent (e.g., 1,4-dioxane, etc.), compound B1 is converted into compound B2 by Suzuki coupling with an aryl or heteroaryl boronic acid (or boronic acid pinacol ester). In the presence of a catalyst (e.g., Pd2(dba)3, etc.), a ligand (e.g., RuPhos, etc.) and a base (e.g., NaO t Compound B2 is converted to Compound B3 by Buchwald-Hartwig coupling with an appropriate amine in the presence of Bu, etc., in a suitable solvent (e.g., PhMe, etc.).

[0190] Option C:

[0191]

[0192] Compound C1 is prepared from 1,2,4,5-tetrazine with a suitable amine in the presence of a suitable base (such as DIEPA, etc.) in a suitable solvent (such as DCM, etc.). Compound C1 is converted to compound C2 by a reverse electron demand Diels-Alder reaction with a suitable enol ether or enamine in a suitable solvent (such as PhMe, etc.). Compound C2 is converted to compound C3 according to the conditions described in Scheme A-Step 2.

[0193] Plan D:

[0194]

[0195] In a suitable solvent (such as THF, etc.), compound D1 is converted into compound D2 (wherein P is a protecting group, such as Me, etc.) by reaction with a suitable organometallic compound (such as Grignard reagent, etc.). In the presence of hydrazine, in a suitable solvent (such as EtOH, etc.), compound D2 is converted into compound D3 by condensation / cyclization sequence. Compound D3 is converted into compound D4 by treatment with a dehydrating halogenating agent (such as POCl3, etc.). In the presence of a catalyst (such as Pd2(dba)3, etc.), a ligand (such as RuPhos, etc.) and a base (such as NaO t In the presence of Bu, etc., in a suitable solvent (e.g., PhMe, etc.), compound D4 is converted to compound D5 by Buchwald-Hartwig coupling with a suitable amine. In a suitable solvent (e.g., DCM, etc.), after treatment with appropriate conditions for removing the protecting group (e.g., BBr in DCM for removing the methyl protecting group), compound D5 is converted to compound D6.

[0196] Plan F:

[0197]

[0198] Compound F1 is converted to compound F2 by reaction with hydrazine in a suitable solvent (e.g., EtOH, etc.). In the presence of a base (e.g., DIPEA, etc.), in a suitable solvent (e.g., DCM, etc.), the reaction of F2 with a chloroformate generates F3, and F3 is treated with a base (e.g., KOH, etc.) in a suitable solvent (e.g., EtOH, etc.) and cyclized at an elevated temperature (e.g., 80° C., etc.) to generate F4. Compound F4 is converted to compound F5 by treatment with POX3 (X=Cl or Br) with or without the addition of a base (e.g., DIPEA, etc.). F5 is treated with a thiolation reagent (e.g., Lawesson reagent (LR) or P2S5) at an appropriate temperature (e.g., 100° C.), and then alkylated with MeI to obtain F6. Compound F6 is converted to F7 by Suzuki coupling with an aryl or heteroboronic acid or boronic ester in the presence of a suitable catalyst (e.g., PdCl2dppf, etc.) and a base (e.g., K2CO3, etc.) in a suitable solvent (e.g., dioxane, etc.). Alternatively, compound F5 is first converted to compound F9 by Suzuki coupling, then sulfided with LR or P2S5, and then alkylated with MeI to convert to F7. F7 is reacted with a nucleophile in a suitable solvent (such as DMSO, etc.) at an elevated temperature (such as 130°C, etc.) to give F8.

[0199] Plan G:

[0200]

[0201] Compound G1 prepared according to step 1 in Scheme F is converted to compound G2 by reacting with a trialkyloxy orthoformate in a suitable solvent (e.g., EtOH, etc.) at an elevated temperature (e.g., 100° C., etc.). G2 is reacted with a halogenating agent (e.g., NBS, etc.) in a suitable solvent (e.g., DMF, etc.) to produce G3, which is reacted with a nucleophilic agent to produce compound G4. Compound G4 is converted to compound G5 by treating with POX3 (X=Cl or Br) with or without the addition of a base (e.g., DIPEA, etc.) at an elevated temperature (e.g., 100° C., etc.). Compound G5 is subjected to Suzuki coupling with an aryl or heteroaryl boronic acid or boronic ester in the presence of a suitable catalyst (e.g., PdCl2dppf, etc.) and a base (e.g., K2CO3, etc.) in a suitable solvent (e.g., dioxane, etc.) to produce G6. Alternatively, compound G4 can be directly converted to compound G6 by BOP-mediated Suzuki coupling.

[0202] Plan H:

[0203]

[0204] The reaction of the metal organic compound with the aldehyde, whether H1 with H2 or H1' with H2', generates alcohol H3. Compound H3 is converted to H4 by treatment with an oxidant (e.g., MnO2, etc.) in a suitable solvent (e.g., DCM, etc.). Alternatively, compound H1" reacts with H2" to directly generate H4. Deprotection of compound H4 gives compound H5. Compound H5 reacts with methyl hydrazinodithiocarboxylate in a suitable solvent (e.g., EtOH, etc.) at an elevated temperature (e.g., 80°C, etc.), and then alkylates with MeI in the presence of a base (e.g., K2CO3, etc.) to give compound H6. Alternatively, compound H5 reacts with hydrazine in a suitable solvent (e.g., EtOH, etc.), and then reacts with chloroformate in the presence of a base (e.g., DIPEA, etc.) in a suitable solvent (e.g., DCM, etc.), and is converted to compound H5' by cyclization treatment with a base (e.g., KOH, etc.) at an elevated temperature (e.g., 80°C, etc.) in a suitable solvent (e.g., EtOH, etc.). SNAr reaction of H6 with a nucleophile in a suitable solvent (eg, DMSO, etc.) at elevated temperature (eg, 130° C., etc.) affords H7.

[0205] Option I:

[0206]

[0207] In the presence of a suitable catalyst (e.g., PdCl2dppf, etc.) and a base (e.g., K2CO3, etc.), in a suitable solvent (e.g., dioxane, etc.), compound I1 is converted to compound I3 by two coupling reactions with boronic acid or boronic ester. Alternatively, compound I4 is converted to compound I5 by coupling with boronic acid or boronic ester, and compound I5 is further converted to compound I3 by BOP-mediated Suzuki coupling with aryl or heteroaryl boronic acid or boronic ester.

[0208] Plan J:

[0209]

[0210] In the presence of a suitable base (e.g., K2CO3, DBU, etc.), in a suitable solvent (e.g., DCE, etc.), compound J1 is converted to compound J2 by ​​reacting with TosMIC. In the presence of a suitable catalyst (e.g., PdCl2dppf, etc.) and a base (e.g., K2CO3, etc.), in a suitable solvent (e.g., dioxane, etc.), compound J2 is subjected to Suzuki coupling with an aryl or heteroaryl boronic acid or boronic ester to obtain J3. Alternatively, compound J1 is converted to compound J5 by coupling with a boronic acid or boronic ester, and compound J5 is further converted to compound J3 by reacting with TosMIC in the presence of a suitable base (e.g., K2CO3, DBU, etc.) in a suitable solvent (e.g., DCE, etc.). J3 is subjected to SNAr reaction with a nucleophile in a suitable solvent (e.g., DMSO, etc.) at an elevated temperature (e.g., 130° C., etc.) to obtain J4.

[0211] Synthesis Example

[0212] In order to describe in more detail and help understand, the following non-limiting examples are provided to more fully illustrate the scope of the compounds described herein, and should not be interpreted as specifically limiting the scope thereof. These variants of the compounds described herein, now known or developed later, will be determined within the scope of those skilled in the art and are considered to fall within the scope of the compounds described herein and required below. These examples illustrate the preparation of certain compounds. It will be appreciated by those skilled in the art that the techniques described in these examples represent techniques described by those of ordinary skill in the art that work well in synthetic practice, and therefore constitute a preferred mode of their practice. However, it should be understood that those skilled in the art should recognize that many changes can be made in the specific methods disclosed according to the present application, and similar or similar results are still obtained without departing from the subject matter and scope of this specification.

[0213] Except for the examples of compounds included below, all numbers used in the specification and claims to represent the quantity of ingredients, reaction conditions, experimental data, etc. should be understood as being modified by the term "about" unless otherwise stated. Therefore, all of these numbers represent approximate values ​​and may vary depending on the desired properties to be obtained by the reaction or due to varying experimental conditions. Therefore, within the expected range of experimental reproducibility, the term "about" in the context of the result data refers to the data range provided, which may vary according to the standard deviation of the mean value. Similarly, for the experimental results provided, the obtained data can be rounded to consistently present the data without losing significant figures. At least it is not intended to limit the application of the doctrine of equivalents to the scope of the claims, and each numerical parameter should be interpreted according to the number of significant figures and the rounding algorithm used by those skilled in the art.

[0214] Although the numerical ranges and parameters setting forth the broad ranges of values ​​in this specification are approximations, the numerical values ​​set forth in the examples listed below are reported as precisely as possible. Any numerical value, however, inherently contains certain errors resulting from the standard deviation found in their respective testing measurements.

[0215] Compound Examples

[0216] As used above and throughout this specification, the following abbreviations, unless otherwise indicated, shall be understood to have the following meanings:

[0217]

[0218]

[0219]

[0220]

[0221] Intermediate 1a: 1-chloro-7-methylpyrrolo[1,2-d][1,2,4]triazine-4(3H)-one

[0222]

[0223] Step 1. 4-Methyl-1H-pyrrole-2-carboxylic acid ethyl ester

[0224] A mixture of ethyl 4-formyl-1H-pyrrole-2-carboxylate (10.3 g, 61.6 mmol, 1.0 equiv) and Pd / C (10%, 2.0 g) in EtOH (150 mL) was stirred at room temperature overnight under hydrogen (balloon). The mixture was filtered. The filtrate was concentrated under reduced pressure. The colorless crude oil (8.50 g, 90% yield) was used in the next step without further purification. MS m / z 154.2 [M+H] + .

[0225] Step 2. 4-Methyl-1H-pyrrole-2-carboxylic acid hydrazide

[0226] A solution of ethyl 4-methyl-1H-pyrrole-2-carboxylate (8.50 g, 55.5 mmol) in N2H4H2O (80%, 28 mL) was heated at 70°C for 45 min. After cooling to room temperature, the mixture was filtered. The filter cake was washed with EtOH and dried in vacuo to give 4-methyl-1H-pyrrole-2-carboxylic acid hydrazide (7.5 g, yield 97%) as a white solid. MS m / z 140.2 [M+H] + ; 1H NMR (400MHz, DMSO-d6) δ 11.07 (s, 1H), 9.14 (s, 1H), 6.62 (s, 1H), 6.55 (s, 1H), 4.27 (s, 2H), 2.00 (s, 3H).

[0227] Step 3. 2-(4-Methyl-1H-pyrrole-2-carbonyl)hydrazine-1-carboxylic acid isobutyl ester

[0228] To a suspension of 4-methyl-1H-pyrrole-2-carboxylic acid hydrazide (7.5 g, 53.9 mmol, 1.0 equiv) and DIEA (20.9 g, 161 mmol, 3.0 equiv) in DCM (540 mL) was added isobutyl phosgene (11.1 g, 80.9 mmol, 1.5 equiv) dropwise and cooled with an ice-water bath. The mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was diluted with DCM and washed with water and brine. The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography, eluted with 0-30% EtOAc hexane solution to give 2-(4-methyl-1H-pyrrole-2-carbonyl)hydrazine-1-carboxylic acid isobutyl ester (12.0 g, 93%) as a white solid. MS m / z 240.2[M+H] + .

[0229] Step 4. 7-Methyl-2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione

[0230] To a solution of 2-(4-methyl-1H-pyrrole-2-carbonyl)hydrazine-1-carboxylic acid isobutyl ester (12.0 g, 50.2 mmol, 1.0 equiv) in EtOH (840 mL) was added KOH (8.40 g, 151 mmol, 3.0 equiv). The mixture was stirred at 85 °C for 2 h. The reaction mixture was cooled to room temperature and filtered. The filter cake was washed with EtOH. The solid was dissolved in water and adjusted to pH = 4 with 1M HCl. The precipitate was filtered. The filter cake was washed with water and dried in vacuo to give 7-methyl-2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione (4.0 g, 48% yield) as a white solid. MS m / z 166.2 [M+H] + .

[0231] Step 5. 1-Chloro-7-methylpyrrolo[1,2-d][1,2,4]triazine-4(3H)-one

[0232] DIEA (3.12 g, 24.2 mmol, 1.0 equiv) was added dropwise to a solution of 7-methyl-2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione (4.0 g, 24.2 mmol, 1.0 equiv) in POCl3 (1.0 M, 25 mL) and cooled with an ice bath. The mixture was stirred at 100°C for 16 h, then cooled to room temperature and concentrated. The residue was carefully poured into ice water and the pH was adjusted to 8 with saturated aqueous NaHCO3. The mixture was extracted with DCM:MeOH (10:1, 100 mL×3), dried over Na2SO4, concentrated, and purified by silica gel column chromatography using 0-50% EtOAc hexane solution to give 1-chloro-7-methylpyrrolo[1,2-d][1,2,4]triazine-4(3H)-one (2.0 g, 45% yield) as a white solid. MS m / z 184.3[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ 12.43 (s, 1H), 7.66 (s, 1H), 6.75 (s, 1H), 2.23 (s, 3H).

[0233] Intermediate 1b: 8-fluoro-2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,,4-dione

[0234]

[0235] Step 1. 3-Fluoro-1H-pyrrole-2-carboxylic acid hydrazide

[0236] A mixture of N2H4·H2O (80% aqueous solution, 18 mL) and 3-fluoro-1H-pyrrole-2-carboxylic acid methyl ester (3.14 g, 20.0 mmol) was stirred at 70°C for 45 minutes. After cooling to room temperature, the precipitate was filtered, and the filter cake was washed with water and dried in vacuo to give 3-fluoro-1H-pyrrole-2-carboxylic acid hydrazide (2.30 g, 73%) as a white solid. MS m / z 144.1 [M+H] + , 1 H NMR (400MHz, DMSO-d6) δ 11.31 (s, 1H), 8.55 (s, 1H), 6.74 (dd, J=4.6, 3.0Hz, 1H), 5.97 (d, J=3.0Hz, 1H), 4.39 (s, 2H).

[0237] Step 2. 2-(3-Fluoro-1H-pyrrole-2-carbonyl)hydrazine-1-carboxylic acid isobutyl ester

[0238] To a solution of 3-fluoro-1H-pyrrole-2-carboxylic acid hydrazide (2.15 g, 15.0 mmol, 1.0 equiv) in DCM (60 mL) was added DIPEA (5.80 g, 45.0 mmol, 3.0 equiv). Isobutyl phosgene (3.00 g, 22.5 mmol, 1.5 equiv) was slowly added to the mixture. The mixture was stirred at room temperature for 16 h. After concentration, the mixture was diluted with EtOAc (200 mL). The organic layer was washed with water and brine, then dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with 0-50% EtOAc hexane solution to give 2-(3-fluoro-1H-pyrrole-2-carbonyl)hydrazine-1-carboxylic acid isobutyl ester (2.44 g, 66.7% yield) as a yellow solid. MS m / z 244.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ11.46 (s, 1H), 9.18 (s, 1H), 9.09 (s, 1H), 6.82 (d, J=4.4Hz, 1H), 6.01 (d, J=2.5Hz, 1H), 3.82 (d, J=6.6Hz, 2H), 2.02-1.67 (m, 1H), 0.92 (d, J=6.7Hz, 6H).

[0239] Step 3. 8-Fluoro-2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione

[0240] To a solution of isobutyl 2-(3-fluoro-1H-pyrrole-2-carbonyl)hydrazine-1-carboxylate (2.44 g, 10.0 mmol, 1.0 eq.) in EtOH (60 mL) was added KOH (1.68 g, 30 mmol, 3 eq.). The mixture was stirred at 85 °C for 2 h. After cooling to room temperature, the mixture was concentrated. The residue was diluted with water (50 mL) and the pH was adjusted to 6-7 with 1 M HCl. The precipitate was filtered, the filter cake was washed with water and dried in vacuo to give a brown solid 8-fluoro-2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione (1.16 g, 68.7% yield). MS m / z 168.0 [MH] - ; 1 H NMR (400MHz, DMSO-d6) δ 11.52 (s, 2H), 7.57 (t, J=4.1Hz, 1H), 6.68 (d, J=3.4Hz, 1H).

[0241] Intermediate 1c: 1-chloro-4-(methylthio)pyrrolo[1,2-d][1,2,4]triazine

[0242]

[0243] Step 1. 1-Chloropyrrolo[1,2-d][1,2,4]triazine-4-ol

[0244] To a stirred solution of 2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione (prepared according to the procedure of Intermediate 1b, 2 g, 13.2 mmol) in POCl3 (60 mL) was added DIEA (1.84 g, 14.2 mmol). The reaction mixture was heated at 100°C for 16 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with cold saturated sodium bicarbonate solution and extracted with dichloromethane (3×50 mL). The combined organic extracts were dried over sodium sulfate and evaporated under reduced pressure. The crude product was further triturated with DCM to give the title compound 1-chloropyrrolo[1,2-d][1,2,4]triazine-4-ol (0.867 g, 38.6% yield) as an off-white solid, which was used in the next step without further purification. MS m / z 169.6 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ12.54 (s, 1H), 7.88-7.87 (m, 1H), 6.92-6.89 (m, 2H).

[0245] Step 2. 1-Chloropyrrolo[1,2-d][1,2,4]triazine-4(3H)-thione

[0246] To a solution of 1-chloropyrrolo[1,2-d][1,2,4]triazine-4-ol (30.0 g, 177 mmol) in toluene (1200 mL) was added Lawesson's reagent (46.2 g, 114 mmol). The reaction was stirred at 120 °C for 16 h. The mixture was diluted with H2O (1.5 L), acidified to pH = 3 with aqueous HCl (1 M), and extracted with EA. The EA layer was washed with saturated aqueous NaHCO3 (1 L) and brine (1 L), then dried over anhydrous Na2SO4 and concentrated to give crude 1-chloro-3H-pyrrolo[1,2-d][1,2,4]triazine-4-thione (crude 35.0 g, excess), which was used in the next step without further purification. . MS m / z 184.1 [MH] - .

[0247] Step 3. 1-Chloro-4-(methylthio)pyrrolo[1,2-d][1,2,4]triazine

[0248] To a solution of 1-chloro-3H-pyrrolo[1,2-d][1,2,4]triazine-4-thione (crude 35.0 g, 189 mmol) in THF (600 mL) and water (300 mL) was added K2CO3 (65.2 g, 472 mmol) and then iodomethane (68.6 g, 483 mmol) at 0°C. The reaction mixture was stirred at room temperature for 2 h, then diluted with water and extracted with EA (3×100 mL). The combined organic layers were washed with water, brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 50 / 1 to 20 / 1) to give the title product 1-chloro-4-methylthio-pyrrolo[1,2-d][1,2,4]triazine (21.8 g, two-step yield 57.9%) as a white solid. MS m / z 200.0 [M+H] + , 1H NMR (400MHz, DMSO-d6) δ: 7.83 (dd, J=2.8Hz, 1.2Hz, 1H), 7.16 (dd, J=4.0Hz, 2.8Hz, 1H), 7.11 (dd, J=4.0Hz, 1.2Hz, 1H), 2.81 (s, 3H).

[0249] The following intermediates were prepared according to the procedure of intermediate 1c by substituting appropriate starting materials, reagents and reaction conditions.

[0250]

[0251] Intermediate 1d: 1-bromo-4-(methylthio)pyrrolo[1,2-d][1,2,4]triazine

[0252]

[0253] Step 1. 1-Bromopyrrolo[1,2-d][1,2,4]triazine-4(3H)-one

[0254] A mixture of 2,3-dihydropyrrolo[1,2-d][1,2,4]triazine-1,4-dione (30.0 g, 0.199 mol, 1.0 equiv) and POBr3 (570 g, 1.99 mol, 10 equiv) was heated at 80°C for 3 h. The hot reaction mixture was slowly poured into a stirred mixture of an ice-saturated aqueous NaHCO3 solution. The mixture was neutralized to pH ~7 with solid NaHCO3 and extracted with a MeOH / DCM mixture (1 L×2, 1:10). The combined organic phases were dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was triturated with DCM, filtered, and washed with DCM. After the solid was dried under vacuum, 1-bromopyrrolo[1,2-d][1,2,4]triazine-4(3H)-one (21.9 g, 51.8% yield) was obtained as a pink solid. MS m / z 214.0, 215.9 [M+H] + .

[0255] Step 2. 1-Bromopyrrolo[1,2-d][1,2,4]triazine-4(3H)-thione

[0256] To a solution of 1-bromopyrrolo[1,2-d][1,2,4]triazine-4(3H)-one (10.0 g, 46.7 mmol, 1.0 equiv) in DMSO (472 mL) was added Lawesson reagent (12.4 g, 30.7 mmol, 0.65 equiv). The reaction mixture was heated at 120°C for 6 hours. The reaction mixture was cooled to room temperature, diluted with saturated aqueous NaHCO3 solution, and extracted with EtOAc (300 mL×3). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluted with 0-30% EtOAc hexane solution to give 1-bromopyrrolo[1,2-d][1,2,4]triazine-4(3H)-thione (1.56 g, yield 14.5%) as a white solid. MS m / z 228.0, 230.0 [MH] - ; 1 H NMR (400Hz, DMSO-d6) δ14.15 (s, 1H), 8.23 ​​(dd, J=2.9, 1.4Hz, 1H), 7.10 (dd, J=3.7, 3.1Hz, 1H), 7.06 (dd, J=3.9, 1.4Hz, 1H).

[0257] Step 3. 1-Bromo-4-(methylthio)pyrrolo[1,2-d][1,2,4]triazine

[0258] To a solution of 1-bromopyrrolo[1,2-d][1,2,4]triazine-4(3H)-thione (2.00 g, 8.69 mmol, 1.0 equiv) and K2CO3 (3.00 g, 21.7 mmol, 2.5 equiv) in THF (43 mL) and water (21 mL) was added iodomethane (1.35 mL, 2.28 g / mL, 21.7 mmol, 2.5 equiv) dropwise at 0°C. The reaction was stirred at 0°C for 1 hour. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL×3). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography using 0-30% EtOAc in hexane to give 1-bromo-4-(methylthio)pyrrolo[1,2-d][1,2,4]triazine (1.40 g, yield 66.1%) as a white solid. MS m / z 243.9, 246.0 [M+H] + ; 1 H NMR (400Hz, DMSO-d6) δ7.83 (dd, J=2.8, 1.0Hz, 1H), 7.15 (dd, J=4.0, 2.8Hz, 1H), 7.03 (dd, J=4.0, 1.0Hz, 1H), 2.79 (s, 3H).

[0259] Intermediate 1e: 1-bromo-4-(methylthio)imidazo[1,5-d][1,2,4]triazine

[0260]

[0261] To a solution of TsMIC (5.21 g, 26.69 mmol, 1.1 eq.) and DBU (4.43 mg, 29.12 mmol, 1.2 eq.) in DCE (50 mL) was added 6-bromo-3-(methylthio)-1,2,4-triazine (5 g, 24.26 mmol, 1.0 eq.) at room temperature. The reaction mixture was stirred for 20 min, then diluted with water (50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=15:1) to give 1-bromo-4-(methylthio)imidazo[1,5-d][1,2,4]triazine (3.71 g, yield 62.38%) as a yellow solid. MS m / z 244.9, 246.9 [M+1] + ; 1H NMR (400MHz, DMSO-d6) δ 8.84 (s, 1H), 7.93 (s, 1H), 2.80 (s, 3H).

[0262] Intermediate 1f: 7-Bromo-2-methylpyrazolo[1,5-d][1,2,4]triazine-4-ol

[0263]

[0264] Step 1. 5-Methyl-1H-pyrazole-3-carboxylic acid hydrazide

[0265] Hydrazine hydrate (54 mL, 1.032 g / mL, 1.11 mol, 6.2 eq) was added to a solution of 5-methyl-1H-pyrazole-3-carboxylic acid methyl ester (25.0 g, 0.178 mol, 1.0 eq) in EtOH (250 mL). The mixture was sealed and heated at 80 °C for 60 h and then cooled to room temperature. The solvent was evaporated in vacuo. The crude product was treated with a 1 / 1 mixture of water / MeOH, filtered, and the filter cake was washed with water. The solid was dried in vacuo to give 5-methyl-1H-pyrazole-3-carboxylic acid hydrazide (21.7 g, 87.0% yield) as a white solid. MS m / z 141.1 [M+H] + .

[0266] Step 2. 2-Methylpyrazolo[1,5-d][1,2,4]triazine-4-ol

[0267] A mixture of 5-methyl-1H-pyrazole-3-carboxylic acid hydrazide (3.00 g, 21.4 mmol, 1.0 equiv) and trimethoxymethane (2.57 mL, 0.970 g / mL, 23.5 mmol, 1.1 equiv) in DMF (10 mL) was sealed in a test tube. The reaction mixture was stirred at 165 ° C for 1 hour under microwave and then cooled to room temperature. The precipitate was collected by filtration, washed with EtOH, and dried in vacuo to give 2-methylpyrazolo[1,5-d][1,2,4]triazine-4-ol (1.70 g, yield 52..9%) as a white solid. MSm / z149.2[MH] - .

[0268] Step 3. 7-Bromo-2-methylpyrazolo[1,5-d][1,2,4]triazine-4-ol

[0269] To a stirred solution of 2-methylpyrazolo[1,5-d][1,2,4]triazine-4-ol (6.80 g, 45.3 mmol, 1.0 equiv) and 2-(tert-butyl)-1,1,3,3-tetramethylguanidine (18.3 mL, 0.85 g / mL, 90.6 mmol, 2.0 equiv) in 1,4-dioxane (290 mL) was partially added benzyltrimethylammonium tribromide (26.5 g, 68.0 mmol, 1.5 equiv) at 10 °C under N2. After the addition was complete, the mixture was warmed to 20 °C and stirred at this temperature for 16 hours. The mixture was then quenched with a mixture of saturated aqueous Na2S2O3 and saturated aqueous NaHCO3, and then extracted with EtOAc (150 mL×3). The combined organic layers were dried over Na2SO4, filtered and concentrated. The crude product was triturated with EtOAc, filtered and washed with EtOAc. The solid was dried under vacuum to give 7-bromo-2-methylpyrazolo[1,5-d][1,2,4]triazine-4-ol (7.60 g, yield 73.6%) as an off-white solid. MS m / z 229.0, 230.9 [M+H] + ; 1 H NMR (400Hz, DMSO-d6) δ12.60 (s, 1H), 7.15 (s, 1H), 2.42 (s, 3H).

[0270] Intermediate 2a: 2-(4-methoxy-2-(2,2,2-trifluoroethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0271]

[0272] Step 1: (2-Bromo-5-methoxyphenyl)methanol

[0273] Borane dimethyl sulfide complex (6.01 mL, 64.6 mmol) was added to an ice-cold solution of 2-bromo-5-methoxybenzoic acid (9.95 g, 43.1 mmol) in THF (86 mL). The resulting mixture was warmed to room temperature and stirred for 16 hours. The reaction mixture was quenched with methanol and stirred for 5 min. The reaction was concentrated and the resulting yellow oil was dissolved in EtOAc (150 mL) and washed with 10% Na2CO3 (30 mL). The aqueous phase was extracted with EtOAc. The combined organic extracts were washed with brine (50 mL), dried (MgSO4) and concentrated to give a colorless oil which was applied to the next step without further purification.

[0274] Step 2: 1-Bromo-2-(bromomethyl)-4-methoxybenzene

[0275] The crude [2-bromo-5-(trifluoromethyl)phenyl]methanol was heated to reflux in concentrated HBr (30 mL) for 2 hours. The reaction was cooled to room temperature and extracted with DCM. The combined organic extracts were washed with saturated NaHCO, brine, dried (Na2SO4), filtered, and concentrated to give a white solid. Purification by SiO2 chromatography (EtOAc: hexane, 10-30%) gave a white solid (10.99 g, 91% for two steps). 1 H NMR (400MHz, CDCl3) δ7.46 (d, J=8.9Hz, 1H), 7.00 (d, J=3.0Hz, 1H), 6.75 (dd, J=8.9Hz, 3.0Hz, 1H), 4.57 (s, 2H), 3.81 (s, 3H).

[0276] Step 3: 1-Bromo-4-methoxy-2-(2,2,2-trifluoroethyl)benzene

[0277] To a solution of 1-bromo-2-(bromomethyl)-4-methoxybenzene (5.62 g, 20.1 mmol) in DMF (45 mL) was added CuI (9.56 g, 50.2 mmol) and argon was passed through the solution. To this solution was added methyl fluorosulfonyl difluoroacetate (6.4 mL, 50.2 mmol) and the resulting reaction mixture was heated at 120°C for 4 h. The reaction mixture was cooled to 0°C, diluted with EtOAc (200 mL) and stirred for 10 min. Concentrated ammonium hydroxide (30 mL) was added dropwise and the mixture was stirred at 0°C for 15 min and then warmed to room temperature. EtOAc (200 mL) and water (100 mL) were added and the layers were separated. . The aqueous layer was extracted with EtOAc (2×100 mL). The combined organic layers were washed with water (100 mL) and brine (100 mL), dried (MgSO4), filtered, and concentrated. Purification by SiO2 chromatography (EtOAc:hexanes, 0-10%) afforded a light yellow oil (4.13 g, 77%). 1 H NMR (400MHz, CDCl3) δ7.50 (d, J=8..9Hz, 1H), 6.93 (d, J=2.8Hz, 1H), 6.78 (dd, J=8.8Hz, 3.1Hz, 1H), 3.81 (s, 3H), 3.60 (q, J=10.5Hz, 2H).

[0278] Intermediate 2b: 4,4,5,5-tetramethyl-2-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,3,2-dioxaborolane

[0279]

[0280] Step 1. (2-Bromo-5-(trifluoromethyl)phenyl)methanol

[0281] To a solution of 2-bromo-5-(trifluoromethyl)benzaldehyde (30.0 g, 118.6 mmol, 1.0 eq.) in MeOH (300 mL) was slowly added NaBH4 (13.5 g, 3 eq.) at 0°C. The mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (250 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The filtrate was concentrated in vacuo to give a white solid (2-bromo-5-(trifluoromethyl)phenyl)methanol (27 g, 78.4% yield). The crude product was used directly without further purification. 1 H NMR (400MHz, CDCl3) δ7.81 (s, 1H), 7.67 (d, J = 8.3Hz, 1H), 7.44-7.39 (m, 1H), 4.81 (s, 2H).

[0282] Step 2. 1-Bromo-2-(bromomethyl)-4-(trifluoromethyl)benzene

[0283] (2-Bromo-5-(trifluoromethyl)phenyl)methanol (26.0 g, 102.0 mmol, 1.0 equiv) was added to aqueous HBr solution (156 mL). Under N2 atmosphere, the reaction mixture was stirred at 120°C for 2 h. The reaction mixture was diluted with NaHCO3 (100 mL) and extracted with DCM (250 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 10%) to give 1-bromo-2-(bromomethyl)-4-(trifluoromethyl)benzene (25 g, yield 76.9%) as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δ 8.08 (d, J=2.0Hz, 1H), 7.92 (d, J=8.4Hz, 1H), 7.65 (dd, J=8.4, 2.1Hz, 1H), 4.83 (s, 2H).

[0284] Step 3. 1-Bromo-2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)benzene

[0285] To a solution of 1-bromo-2-(bromomethyl)-4-(trifluoromethyl)benzene (40.5 g, 127.4 mmol, 1.0 equiv) in DMF (280 mL) was added CuI (60.7 g, 2.5 equiv) and ethyl 2,2-difluoro-2-(fluorosulfonyl)acetate (65.7 g, 2.5 equiv) under N2 and room temperature. The reaction mixture was stirred overnight at 120 ° C in a N2 atmosphere. The reaction mixture was cooled to 0 ° C, diluted with EtOAc (1200 mL) and stirred at 0 ° C for 10 minutes. Ammonium hydroxide solution (240 mL) was added dropwise. The mixture was warmed from 0 ° C to room temperature within 20 minutes under stirring. The reaction mixture was diluted with water (500 mL) and extracted with EtOAc (550 mL×3). The combined organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 10%) to give 1-bromo-2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)benzene (16.8 g, yield 17.9%) as a yellow oil. 1 HNMR (400MHz, DMSO-d6) δ7.95 (dd, J=15.1, 8.2Hz, 2H), 7.70 (dd, J=8.4, 2.1Hz, 1H), 3.97 (q, J=11.0Hz, 2H).

[0286] Step 4. 4,4,5,5-Tetramethyl-2-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,3,2-dioxaborolane

[0287] To a solution of 1-bromo-2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)benzene (16.8 g, 54.7 mmol, 1.0 eq.) in 1,4-dioxane (160 mL) was added 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (27.8 g, 2 eq.), Pd(dppf)Cl2 (4.0 g, 0.1 eq.) and KOAc (16.1 g, 3 eq.) under N2 at room temperature. The reaction mixture was stirred at 100°C under N2 for 12 h. The reaction mixture was diluted with water (90 mL) and extracted with EtOAc (150 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 10%) to give 4,4,5,5-tetramethyl-2-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,3,2-dioxaborolane (10.5 g, 45.8% yield) as a yellow oil. 1H NMR (400MHz, DMSO-d6) δ7.95 (d, J=7.8Hz, 1H), 7.79-7.73 (m, 2H), 4.08 (q, J=11.2Hz, 2H), 1.32 (s, 12H).

[0288] The following intermediates were prepared according to the procedures of Intermediates 2a and 2b by substituting appropriate starting materials, reagents and reaction conditions.

[0289]

[0290] Intermediate 2c: 2-[2-(difluoromethyl)-4-(trifluoromethyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0291]

[0292] Step 1. 1-Bromo-2-(difluoromethyl)-4-(trifluoromethyl)benzene

[0293] DAST (10 equivalents, 39.5 mmol) was added dropwise to a -78°C solution of 2-bromo-5-(trifluoromethyl)benzaldehyde (1 g, 3.9 mmol) in DCM (5 mL, 78.0 mmol). The reaction was stirred for 15 min and then warmed to room temperature. After 4 hours, TLC showed incomplete conversion, so the mixture was cooled to -78°C again and additional DAST (1 equivalent, 3.9 mmol) was added. The mixture was warmed to room temperature and stirred overnight, then poured into ice-cold NH4OH and extracted twice with DCM. The combined organic extracts were washed with brine and dried over sodium sulfate. The solvent was evaporated in vacuo to give a residue, which was purified with a short plug of silica gel to give a crude product, colorless oily 1-bromo-2-(difluoromethyl)-4-(trifluoromethyl)benzene (350 mg, 1.2 mmol, 32.2% yield).

[0294] Step 2. 2-[2-(Difluoromethyl)-4-(trifluoromethyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0295] To a solution of 1-bromo-2-(difluoromethyl)-4-(trifluoromethyl)benzene (100 mg, 0.36 mmol) in 1,4-dioxane (2 mL) was added pinacol diboron (1.5 eq., 0.5 mmol), potassium acetate (2 eq., 0.72 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (0.15 eq., 0.05 mmol). The mixture was stirred at 100 °C under N2 atmosphere for 16 h and monitored by TLC and LCMS. After the reaction was complete, the solvent was removed in vacuo and the residue was purified by column chromatography (10-15% EA in PE) to give 2-[2-(difluoromethyl)-4-(trifluoromethyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (30 mg, 0.09 mmol, 25.6% yield) as a colorless oil. 1 H NMR (400MHz, DMSO-d6) δ 8.00 (d, J=7.6Hz, 1H), 7.94 (d, J=8.4Hz, 1H), 7.92 (s, 1H), 7.38 (t, J=55.6Hz, 1H), 1.34 (s, 12H).

[0296] Intermediate 2d: 2-(4-chloro-2-(difluoromethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0297]

[0298] Following a similar procedure to that of Intermediate 2c, substituting 2-bromo-5-chlorobenzaldehyde for 2-bromo-5-(trifluoromethyl)benzaldehyde in Step 1, the title compound was prepared. 1 H NMR (400MHz, DMSO-d6) δ7.79 (d, J=2.0Hz, 1H), 7.68-7.63 (m, 2H), 7.32 (t, J=55.6Hz, 1H), 1.32 (s, 12H).

[0299] Intermediate 2e: 2-(2-(difluoromethoxy)-4-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0300]

[0301] Step 1. 1-Bromo-2-(difluoromethoxy)-4-(trifluoromethyl)benzene

[0302] A mixture of 2-bromo-5-(trifluoromethyl)phenol (5.00 g, 20.7 mmol, 1.0 equiv), sodium 2-chloro-2,2-difluoroacetate (7.07 g, 51.9 mmol, 2.50 equiv) and cesium carbonate (11.5 g, 41.5 mmol, 2.0 equiv) in water (20 mL) and N,N-dimethylformamide (80 mL) was heated at 100 ° C for 12 hours. After the reaction was completed, the reaction mixture was cooled to room temperature and diluted with EtOAc. The reaction mixture was washed with water and brine. The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography eluted with 0-50% EtOAc hexane solution to give 1-bromo-2-difluoromethoxy-4(trifluoromethyl)benzene (2.8 g, 47% yield) as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ 8.01 (d, J=8.4Hz, 1H), 7.69 (s, 1H), 7.46 (t, J=72.8Hz, 1H), 7.5 (d, J=8.4Hz, 1H).

[0303] Step 2. 2-(2-(difluoromethoxy)-4-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0304] To a solution of 1-bromo-2-difluoromethoxy-4-trifluoromethylbenzene (2.80 g, 9.62 mmol) in 1,4-dioxane (30 mL) was added diboric acid pinacol ester (36.6 g, 14.4 mmol, 1.50 equiv), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (704 mg, 0.96 mmol, 0.1 equiv), potassium acetate (2.82 g, 28.9 mmol, 3.0 equiv). The reaction mixture was heated at 100 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with EA, washed with water and brine. The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with n-hexane to give 2-(2-(difluoromethoxy)-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane as a white solid (1.70 g, yield 52%). 1 H NMR (400MHz, DMSO-d6) δ7.87 (d, J=7.6Hz, 1H), 7.66 (d, J=7.6Hz, 1H), 7.50 (s, 1H), 7.24 (t, J=74.0Hz, 1H), 1.31 (s, 12H).

[0305] Intermediate 2f: 2-(2,4-bis(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0306]

[0307] Step 1. 1-Bromo-2,4-bis(difluoromethoxy)benzene

[0308] At 0°C, KOH (60.0 g, 1.07 mol, 20 equiv) was slowly added to a solution of 4-bromobenzene-1,3-diol (10.0 g, 53.4 mmol, 1.0 equiv) in ACN / H2O (1:1, 100 mL). The mixture was stirred for 10 min and then cooled to -10°C. Diethyl (bromodifluoromethyl)phosphonate (42.70 g, 160.2 mmol, 3.0 equiv) was slowly added, the mixture was stirred for 10 min, then warmed to room temperature and stirred for 2 h. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 5%) to give 1-bromo-2,4-bis(difluoromethoxy)benzene (3.30 g, yield 21.3%) as a yellow oil. 1 HNMR (400MHz, DMSO-d6) δ7.84 (d, J=8.8Hz, 1H), 7.38 (t, J=72.0Hz, 1H), 7.35 (t, J=76.0Hz, 1H), 7.26 (d, J=2.8Hz, 1H), 7.11 (dd, J=8.8, 2.8Hz, 1H).

[0309] Step 2. 2-(2,4-bis(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0310] To a solution of 1-bromo-2,4-bis(difluoromethoxy)benzene (3.30 g, 11.4 mmol, 1.0 equiv) in 1,4-dioxane was added KOAc (2.23 g, 22.8 mmol, 2.0 equiv), Pd(dppf)Cl2 (0.830 g, 1.10 mmol, 0.1 equiv) and pinacol diboron (4.34 g, 17.1 mmol, 1.5 equiv). The reaction mixture was stirred at 100°C under N2 for 16 h. The mixture was filtered and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 10%) to give 2-(2,4-bis(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.0 g, 52% yield) as a yellow oil. 1H NMR (400 MHz, methanol-d4) δ 7.72 (d, J = 8.0 Hz, 1H), 7.36 (t, J = 73.2 Hz, 1H), 7.12 (t, J = 74.4 Hz, 1H), 7.12-7.08 (m, 1H), 7.02-6.97 (m, 1H), 1.29 (s, 12H).

[0311] Intermediate 2g. 2-(4-bromo-2-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0312]

[0313] Step 1. 4-Bromo-2-(difluoromethoxy)-1-nitrobenzene

[0314] To a solution of 5-bromo-2-nitrophenol (5.7 g, 26.15 mmol) in DMF (135 mL) / H2O (15 mL) were added Cs2CO3 (17.0 g, 52.29 mmol) and sodium 2-chloro-2,2-difluoroacetate (10.0 g, 65.37 mmol). The mixture was stirred at room temperature for 15 minutes and then heated to 100°C for 16 hours. The reaction mixture was cooled to room temperature and partitioned between water (500 mL) and EA (200 mL). The aqueous phase was extracted with EA (2×200 mL). The combined organic extracts were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column (0-20% EA in PE) to give 4-bromo-2-(difluoromethoxy)-1-nitrobenzene (4.0 g, 57% yield) as a yellow oil. 1H NMR (400MHz, DMSO-d6) δ 8.05 (d, J=8.8Hz, 1H), 7.83 (d, J=1.6Hz, 1H), 7.74 (dd, J=8.8, J=1.6Hz, 1H), 7.43 (t, J=72.4Hz, 1H).

[0315] Step 2. 4-Bromo-2-(difluoromethoxy)aniline

[0316] To a solution of 4-bromo-2-(difluoromethoxy)-1-nitrobenzene (1.0 g, 3.73 mmol) in THF (10 mL) / H2O (5 mL) was added a solution of NH4Cl (1.6 g, 29.85 mmol) in H2O (10 mL) and Zn (1.95 g, 29.85 mmol). The mixture was stirred at room temperature for 2 h, then filtered through a celite pad and rinsed with EA. The filtrate was extracted with EA (30 mL×2). The organic extract was dried over Na2SO4, decanted, concentrated, and purified on a silica gel column (0-10% EA in PE) to give 4-bromo-2-(difluoromethoxy)aniline (750 mg, 84% yield) as a yellow oil. MS m / z 236.0[M-1] - ; 1H NMR (400MHz, DMSO-d6) δ7.15 (s, 1H), 7.11 (dd, J=8.8, 2.0Hz, 1H), 7.09 (t, J=74.0Hz, 1H), 6.73 (d, J=8.8Hz, 1H), 5.28 (s, 2H).

[0317] Step 3. 2-(4-Bromo-2-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0318] To a solution of 4-bromo-2-(difluoromethoxy)aniline (13.0 g, 54.61 mmol) in MeCN (300 mL) was added t-BuNO2 (8.45 g, 81.92 mmol), (BPin)2 (15.26 g, 60.08 mmol) at 0° C. The mixture was stirred at 80° C. for 4 h, then concentrated and purified by silica gel column (0-5% EA in PE) to give 2-(4-bromo-2-(difluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.4 g, 28% yield) as a yellow oil. 1H NMR (400MHz, DMSO-d6) δ7.60 (d, J=8.0Hz, 1H), 7.51 (dd, J=8.0, J=1.6Hz, 1H), 7.42 (s, 1H), 7.15 (t, J=74.2Hz, 1H), 1.29 (s, 12H).

[0319] The following intermediates were prepared according to the procedures of intermediates 2e-2g by substituting appropriate starting materials, reagents and reaction conditions.

[0320]

[0321] Intermediate 2h: 2-(2-cyclopropyl-6-fluoro-4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0322]

[0323] Step 1. 3-Cyclopropyl-5-fluorophenol

[0324] To a solution of 3-bromo-5-fluorophenol (5.0 g, 26.3 mmol, 1.0 equiv) in dioxane (40 mL) and water (10 mL) was added cyclopropylboronic acid (4.5 g, 52.6 mmol, 2.0 equiv), Pd(dppf)Cl2 (377 mg, 0.52 mmol, 0.2 equiv) and cesium carbonate (25.7 g, 78.9 mmol, 3.0 equiv). The resulting mixture was stirred under nitrogen at 100°C for 16 hours, then diluted with ethyl acetate (200 mL) and washed with water (200 mL). The aqueous phase was adjusted to pH ~4 by the addition of 6M HCl and then extracted with ethyl acetate (3×100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (10% ethyl acetate in petroleum ether) to give 4-bromo-3-cyclopropyl-5-fluorophenol (3.1 g, yield 77.5%) as a yellow oil. 1H NMR (400 MHz, methanol-d4) δ 6.31 (t, J = 1.8 Hz, 1H), 6.28-6.25 (m, 1H), 6.25-6.22 (m, 1H), 1.85-1.76 (m, 1H), 0.97-0.88 (m, 2H), 0.66-0.58 (m, 2H).

[0325] Step 2. 4-Bromo-3-cyclopropyl-5-fluorophenol

[0326] To a solution of 4-bromo-3-cyclopropyl-5-fluorophenol (3 g, 19.7 mmol, 1.0 equiv) in dichloromethane (15 mL) and methanol (15 mL) was added tetrabutylammonium tribromide (10.4 g, 21.7 mmol, 1.1 equiv). The resulting mixture was stirred at room temperature for 2 h. Water (100 mL) was then added. The mixture was extracted with dichloromethane (3×50 mL). The organic phase was washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (10% ethyl acetate in petroleum ether) to give 4-bromo-3-cyclopropyl-5-fluorophenol (3.9 g, 85.7% yield) as a yellow oil. 1H NMR (400 MHz, methanol-d4) δ 6.50-6.42 (m, 1H), 6.27-6.20 (m, 1H), 2.23-2.09 (m, 1H), 1.03-0.97 (m, 2H), 0.66-0.59 (m, 2H).

[0327] Step 3. 2-Bromo-1-cyclopropyl-3-fluoro-5-methoxybenzene

[0328] To a solution of 4-bromo-3-cyclopropyl-5-fluorophenol (3.9 g, 16.9 mmol, 1.0 equiv) in N,N-dimethylformamide (40 mL) was added iodomethane (1.58 mL, 25.4 mmol, 1.5 equiv) and potassium carbonate (4.67 g, 33.8 mmol, 2.0 equiv). The resulting mixture was stirred at 70 °C for 2 h. The reaction mixture was diluted with ethyl acetate (150 mL) and washed with water (3×100 mL) and brine (100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (3% ethyl acetate in petroleum ether) to give 2-bromo-1-cyclopropyl-3-fluoro-5-methoxybenzene (3.1 g, 74.8% yield) as a pink oil. 1H NMR (400 MHz, methanol-d4) δ 6.69-6.55 (m, 1H), 6.48-6.28 (m, 1H), 3.87-3.72 (m, 3H), 2.26-2.08 (m, 1H), 1.03-0.93 (m, 2H), 0.77-0.59 (m, 2H).

[0329] Step 4. 2-(2-Cyclopropyl-6-fluoro-4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0330] At -78 ° C, 2-bromo-1-cyclopropyl-3-fluoro-5-methoxybenzene (3.0 g, 12.2 mmol, 1.0 equiv) was added to a solution of tetrahydrofuran (30 mL) with n-butyl lithium (6.3 mL, 15.9 mmol, 1.3 equiv, 2.5 M hexane solution). The resulting mixture was stirred at -78 ° C for 30 min, and then 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborane (2.97 mL, 14.6 mmol, 1.2 equiv) was added dropwise. The resulting mixture was warmed to room temperature and stirred for another 2 h. The reaction was quenched with water (100 mL), and the mixture was extracted with ethyl acetate (4×50 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (3% ethyl acetate in petroleum ether) to give 2-(2-cyclopropyl-6-fluoro-4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.9 g, yield 53.3%) as a light yellow oil. MS m / z 293.2 [M+H] +; 1H NMR (400 MHz, methanol-d4) δ 6.52-6.37 (m, 1H), 6.31-6.17 (m, 1H), 3.79-3.69 (m, 3H), 2.34-2.19 (m, 0.8H), 2.00-1.90 (m, 0.16H), 1.39-1.33 (m, 12H), 0.95-0.84 (m, 2H), 0.69-0.60 (m, 2H).

[0331] Intermediate 2i. 2-(4-methoxy-2-(2,2,2-trifluoroethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0332]

[0333] Step 1. 1-Bromo-4-methoxy-2-(2,2,2-trifluoroethoxy)benzene

[0334] To a solution of NaH (2.2 g, 53.6 mmol, 60% in oil, 2.2 equiv) in DMF (20 mL) at 0°C under N2 was added 2,2,2-trifluoroethane-1-ol (4.9 g, 48.8 mmol, 2.0 equiv) dropwise. The mixture was stirred at 0°C under N2 for 0.5 h, then 1-bromo-2-fluoro-4-methoxybenzene (5.0 g, 24.4 mmol, 1.0 equiv) was added. The mixture was warmed to 100°C, stirred under N2 for 16 h, then poured into water (100 mL) and extracted with dichloromethane (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (0-7% ethyl acetate in petroleum ether) to give 1-bromo-4-methoxy-2-(2,2,2-trifluoroethoxy)benzene (3.5 g, 12.3 mmol, 50.4% yield) as a colorless oil. 1H NMR (400 MHz, DMSO-d6) δ 7.49 (d, J = 8.8 Hz, 1H), 6.85 (d, J = 2.8 Hz, 1H), 6.61 (dd, J = 8.8, 2.8 Hz, 1H), 4.86 (q, J = 8.8 Hz, 2H), 3.78 (s, 3H).

[0335] Step 2. 2-(4-methoxy-2-(2,2,2-trifluoroethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,,2-dioxaborolane

[0336] To a solution of 1-bromo-4-methoxy-2-(2,2,2-trifluoroethoxy)benzene (3.50 g, 12.3 mmol, 1.0 equiv) in dioxane (40 mL) was added 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (3.7 g, 14.7 mmol, 1.2 equiv), Pd(dppf)Cl2 (898.4 mg, 1.2 mmol, 0.1 equiv) and KOAc (3.6 g, 36.8 mmol, 3.0 equiv). The resulting mixture was stirred at 100 °C under nitrogen for 16 hours. It was then cooled, diluted with ethyl acetate (200 mL), washed with water (200 mL), and extracted with ethyl acetate (3 x 100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography (0-10% ethyl acetate in petroleum ether) to give 2-(4-methoxy-2-(2,2,2-trifluoroethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.5 g, 4.5 mmol, yield 36.8%) as a white solid. MS m / z 333.1 [M+H] + ; 1H NMR (400 MHz, methanol-d4) δ 7.56 (d, J = 8.4 Hz, 1H), 6.63 (dd, J = 8.0, 2.0 Hz, 1H), 6.54 (d, J = 2.0 Hz, 1H), 4.47 (q, J = 8.4 Hz, 2H), 3.81 (s, 3H), 1.32 (s, 12H).

[0337] Intermediate 2j. 2-(4-methoxy-2-(trifluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0338]

[0339] Step 1. 4-iodo-2-(trifluoromethoxy)aniline

[0340] To a solution of 2-(trifluoromethoxy)aniline (29 g, 163.73 mmol, 1.0 equiv) in acetic acid (80 mL) was slowly added NIS (40.52 g, 180.10 mmol, 1.1 equiv) at 0°C. The mixture was stirred at room temperature for 3 h. The reaction solution was concentrated in vacuo to remove acetic acid, and then the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (EA in PE = 0%-20) to give 4-iodo-2-(trifluoromethoxy)aniline (40.6 g, yield 81.8%) as a red solid. MS m / z 303.9 [M+1]+ ; 1 H NMR (400MHz, DMSO-d6) δ7.35-7.32 (m, 2H), 6.66 (d, J=8.4Hz, 1H), 5.59 (s, 2H).

[0341] Step 2. 4-Methoxy-2-(trifluoromethoxy)aniline

[0342] To a solution of 4-iodo-2-(trifluoromethoxy)aniline (23.8 g, 78.54 mmol, 1.0 equiv) in MeOH (200 mL) was added 1,10-phenanthroline (1.42 g, 7.85 mmol, 0.1 equiv), Cs2CO3 (51.18 g, 157.08 mmol, 2.0 equiv) and CuI (0.748 g, 3.93 mmol, 0.05 equiv) at room temperature under N2. The reaction mixture was stirred at 110°C for 16 h in an autoclave reactor. The reaction solution was concentrated under vacuum to remove MeOH, and then the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (250 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (EA in PE = 0% to 10%) to give 4-methoxy-2-(trifluoromethoxy)aniline (11 g, yield 67.61%) as a yellow oil. MS m / z 208.2 [M+1] + ; 1 H NMR (400 MHz, methanol-d4) δ 6.91-6.67 (m, 3H), 3.71 (s, 3H).

[0343] Step 3. 2-(4-methoxy-2-(trifluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0344] At 0°C, tBuNO2 (8.21 g, 79.65 mmol, 1.5 eq.) and (BPin)2 (16.18 g, 63.72 mmol, 1.2 eq.) were added to a solution of 4-methoxy-2-(trifluoromethoxy)aniline (11 g, 53.10 mmol, 1.0 eq.) in ACN (25 mL). The reaction mixture was stirred at 80°C under N2 atmosphere for 4 h, then cooled and concentrated in vacuo, diluted with water (200 mL) and extracted with ethyl acetate (150 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (EA in PE = 0% to 2%) to give 2-(4-methoxy-2-(trifluoromethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.6 g, yield 33.15%) as a yellow oil. MS m / z 319.0 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δ7.71 (d, J = 8.4Hz, 1H), 7.08-7.03 (m, 1H), 6.92 (s, 1H), 3.87 (s, 3H), 1.32 (s, 12H).

[0345] Intermediate 3a: (3R)-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]piperidin-3-amine

[0346]

[0347] Step 1: (3R)-tert-butyl 3-(benzyloxycarbonylamino)piperidine-1-carboxylate

[0348] To a solution of (R)-tert-butyl 3-aminopiperidine-1-carboxylate (30 g, 0.15 mol) in THF (330 mL) and water (83 mL) was added Na2CO3 (43 g, 0.31 mol). Cbz-Cl (34, 0.199 mol) was then added dropwise to the mixture under ice bath conditions and monitored by TLC. After 5 hours, the reaction mixture was extracted with EA, dried over Na2SO4, and evaporated in vacuo. The crude product was purified by flash chromatography (PE:EA=4:1) to give (3R)-tert-butyl 3-(benzyloxycarbonylamino)piperidine-1-carboxylate (35 g, yield 84%). MS m / z 235.1[M-Boc+H] + .

[0349] Step 2. Benzyl N-[(3R)-3-piperidinyl]carbamate

[0350] To a solution of tert-butyl (3R)-3-(benzyloxycarbonylamino)piperidine-1-carboxylate (35 g, 0.104 mol) in 450 mL of DCM was added HCl (350 mL, 4 M in 1,4-dioxane) and monitored by TLC. After 2 h, the solvent was removed under vacuum to give crude N-[(3R)-3-piperidinyl]carbamic acid benzyl ester (25 g, 100% yield), which was used in the next step without purification. MS m / z 234.1 [M+H] + .

[0351] Step 3. Benzyl N-[(3R)-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-piperidinyl]carbamate

[0352] To a solution of benzyl N-[(3R)-3-piperidinyl]carbamate (25 g, 0.106 mol) in CH3CN (680 mL) was added Cs2CO3 (175 g, 0.537 mol) and (2-bromoethoxy)(tert-butyl)dimethylsilane (40 g, 0.167 mol). The reaction mixture was stirred at 90°C for 16 h, then filtered and the solvent removed in vacuo. The crude product was purified by flash chromatography to give the product benzyl N-[(3R)-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-piperidinyl]carbamate (22.5 g, 53% yield). MS m / z 393.2 [M+H] + .

[0353] Step 4. (3R)-1-[2-[tert-Butyl(dimethyl)silyl]oxyethyl]-piperidin-3-amine

[0354] Pd / C (4.5 g, 20%) was added to a solution of benzyl N-[(3R)-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-3-piperidinyl]carbamate (22.5 g, 0.057 mol) in MeOH (60 mL). The system was evacuated and hydrogen was re-injected. The mixture was then stirred at room temperature overnight. After the reaction was completed, the mixture was filtered, the solvent was removed in vacuo, and the crude product was purified by flash chromatography to give (3R)-1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]piperidin-3-amine (12 g, 81% yield). MS m / z 259.2[M+H] + . 1H NMR (400MHz, CDCl3) δ3.68 (t, J=6.6Hz, 2H), 2.85-2.74 (m, 2H), 2.67-2.58 (m, 1H), 2.46 (t, J=6.4Hz, 2H), 2.11-2.01 (m, 1H), 1.94- 1.83(m,1H),1.83-1.71(m,4H),1.68-1.58(m,1H),1.56-1.43(m,1H),1.09-0.96(m,1H),0.88-0.79(m,9H),0.04-(-)0.04(m,6H).

[0355] Intermediate 3b: (R)-1-ethylpiperidin-3-amine hydrochloride

[0356]

[0357] Step 1: (R)-tert-butyl (1-ethylpiperidin-3-yl)carbamate

[0358] At 20 ℃, N2, K2CO3 (2.76g, 20mmol) and MeI (3.12g, 20mmol) were added to a solution of (R)-piperidin-3-ylcarbamic acid tert-butyl ester (4g, 20mmol) in MeCN (40mL). The mixture was stirred at 20 ℃ for 16 hours. TLC showed that the reaction was complete. The reaction mixture was poured into water (100mL), and extracted with EA (100mL×2). The combined organic layer was washed with brine (100mL×2), dried over Na2SO4, filtered and concentrated. Purified by column (DCM:MeOH=0-10%), yellow oily (R)-(1-ethylpiperidin-3-yl)carbamic acid tert-butyl ester (3.6g, 15.8mmol, yield 78.9%) was obtained. 1 H NMR (400MHz, DMSO-d6) δ6.65 (d, J=7.8Hz, 1H), 3.32 (s, 1H), 2.70 (dd, J=41.0, 9.7Hz, 2H), 2.29 (q, J=7.1Hz, 2H), 1.83-1.53 ​​(m, 4H), 1.48-1.40 (m, 1H), 1.37 (s, 9H), 1.11 (qd, J=11.8, 3.7Hz, 1H), 0.96 (t, J=7.2Hz, 3H).

[0359] Step 2: (R)-1-Ethylpiperidin-3-amine hydrochloride

[0360] To a solution of (R)-tert-butyl (1-ethylpiperidin-3-yl)carbamate (3.6 g, 15.8 mmol) in MeOH (10 mL) was added HCl (30 mL, 3 M in EA) at room temperature. The mixture was stirred at 20 °C for 16 hours. TLC showed that the reaction was complete. The reaction mixture was concentrated to give (R)-1-ethylpiperidin-3-amine hydrochloride (3.03 g, 15.0 mmol, 95.0% yield) as a yellow oil. MS m / z 129.2 [M+H] + ; 1 H NMR (400 MHz, methanol-d4) δ 3.83-3.59 (m, 3H), 3.36-3.30 (m, 2H), 3.18-2.96 (m, 2H), 2.25 (d, J = 12.7 Hz, 1H), , 2.15 (d, J = 14.9 Hz, 1H), 2.07-1.92 (m, 1H), 1.76-1.72 (m, 1H), 1.43 (t, J = 7.3 Hz, 3H).

[0361] Intermediate 9c: (R)-1-(3-aminopiperidin-1-yl)-2-methyl-2-propanol

[0362]

[0363] Step-1: (R)-tert-butyl (1-(2-hydroxy-2-methylpropyl)piperidin-3-yl)carbamate

[0364] A mixture of (R)-tert-butyl piperidin-3-ylcarbamate (700 mg, 3.5 mmol, 1.0 equiv) and 2,2-dimethyloxirane (800 mg, 11.1 mmol, 3.2 equiv) was stirred at 90 °C for 18 h. The crude reaction mixture was cooled to room temperature and diluted with DCM (20 mL). The organic phase was washed with saturated aqueous NaHCO3 (4 mL x 2), then with water and brine. The organic phase was dried over MgSO4, filtered, and concentrated in vacuo. The crude product was then used in the following step. MS m / z 273.3 [M+H] + .

[0365] Step-2: (R)-1-(3-aminopiperidin-1-yl)-2-methyl-2-propanol

[0366] The crude compound obtained in the previous step was dissolved in DCM (5 mL), and then 2M HCl in ether (10 mL) was slowly added at room temperature while the mixture was vigorously stirred. The reaction mixture was stirred at room temperature overnight and concentrated in vacuo to give the hydrochloride salt as a light brown solid (520 mg). The amine was used without further purification. MS m / z 173.2 [M+H] + .

[0367] The following intermediates were prepared according to the procedures of intermediate 3b-c or c by substituting appropriate starting materials, reagents and reaction conditions.

[0368]

[0369] Intermediate 3d: (R)-1-(2-(difluoromethoxy)ethyl)piperidin-3-amine hydrochloride

[0370]

[0371] Step 1: 2-Hydroxyethyl 4-methylbenzenesulfonate

[0372] A mixture of ethylene glycol (2.5 g, 40 mmol), TsCl (1.90 g, 10 mmol), pyridine (1.1 g, 12 mmol) and DMAP (12 mg, 0.1 mmol) was stirred at room temperature for 18 h. The reaction mixture was partitioned between DCM (20 mL) and 0.5 M hydrochloric acid. The organic layer was dried (Na2SO4), filtered and concentrated. Chromatographic purification (EtOAc: hexane, 0-60%) was performed on SiO2 to give a colorless oil (1.32 g, 78%). 1 H NMR (400MHz, CDCl3) δ7.82 (d, J=8.25Hz, 2H), 7.37 (d, J=8.13Hz, 2H), 4.15 (t, J=4.50Hz, 2H), 3.83 (t, J=4.63Hz, 2H), 2.46 (s, 3H), 2.00 (br s, 1H).

[0373] Step 2. 2-(Difluoromethoxy)ethyl 4-methylbenzenesulfonate

[0374] Copper (I) iodide (0.300 g, 1.57 mmol) was added to a stirred solution of 2-hydroxyethyl 4-methylbenzenesulfonate (1.70 g, 7.86 mmol) in MeCN (13 mL). The resulting mixture was stirred at 70 ° C, and a solution of 2,2-difluoro-2-fluorosulfonylacetic acid (2.80 g, 15.7 mmol) in MeCN (10 mL) was added dropwise over a 25 min period. The reaction mixture was treated with anhydrous Na2SO4 (small spoon) and continued to stir for 1.5 h. The mixture was then cooled to room temperature, diluted with Et2O, and washed with brine, a 1:1 mixture of brine: water (2×) and brine. The organic phase was dried (Na2SO4), filtered and concentrated. Purification by chromatography on SiO2 (EtOAc: hexane, 0-25%) gave a pale yellow oil (0.759 g, 36%). 1H NMR (400MHz, CDCl3) δ7.79 (d, J=8.25Hz, 2H), 7.49 (d, J=8.13Hz, 2H), 6.65 (t, J=75.04Hz, 1H), 4.22-4.16 (m, 2H), 4.04-3.98 (m, 2H), 2.43 (s, 3H)

[0375] Steps 3 and 4: (R)-1-(2-(difluoromethoxy)ethyl)piperidin-3-amine hydrochloride

[0376] 4-Methylbenzenesulfonic acid-2-(difluoromethoxy)ethyl ester (0.764 g, 2.87 mmol) and K2CO3 (0.690 g, 4.99 mmol) were added to a solution of tert-butyl N-[(3R)-3-piperidinyl]carbamate (0.500 g, 2.50 mmol) in DMF (10 mL). The mixture was stirred at 100 ° C for 2 h, then diluted with DCM / iPrOH (9:1), washed with water, brine, dried over Na2SO4, filtered and concentrated. Purification by chromatography on SiO2 (EtOAc: hexane, 5-60%) gave a light yellow oil (0.709 g). The oil was dissolved in MeOH (2.0 mL), treated with HCl / dioxane (4.0 M, 3 mL) and stirred for 4 h. The mixture was concentrated, resuspended in ether, and filtered to give a white solid (0.371 g, 67%). 1 H NMR (400MHz, D2O) δ6.45 (t, J=73.6Hz, 1H), 4.26 (t, J=5.2Hz, 2H), 3.81 (d, J=11.2Hz, 1H), 3.67 -3.58 (m, 2H), 3.55 (t, J=4.8Hz, 2H), 3.15-3.02 (m, 2H), 2.21 (d, J=12.4Hz, 1H), 2.11 (d, J=15.2Hz, 1H), 1.90-1.79 (m, 1H), 1.70-1.60 (m, 1H).

[0377] Intermediate 3f: (3R,5R)-5-Fluoro-1-methylpiperidin-3-amine hydrochloride

[0378]

[0379] Step 1. Methyl (2S,4S)-4-hydroxypyrrolidine-2-carboxylate

[0380] 1-(tert-Butyl)-2-methyl (2S,4S)-4-hydroxypyrrolidine-1,2-dicarboxylate (10.0 g, 40.8 mmol) was dissolved in DCM (50 mL) and cooled to 0°C. TFA (10 mL, 132.3 mmol) was slowly added, the mixture was warmed to room temperature and stirred for 1 h. The mixture was concentrated under vacuum to give crude (2S,4S)-4-hydroxypyrrolidine-2-carboxylic acid methyl ester as a yellow oil, which was used in the next step without further purification. MS m / z 146.1 [M+H] + .

[0381] Step 2. Methyl (2S,4S)-4-hydroxy-1-tritylpyrrolidine-2-carboxylate

[0382] To the solution of crude (2S,4S)-4-hydroxypyrrolidine-2-carboxylic acid methyl ester in DCM (50 mL) in the above step, TEA (12.4 g, 123 mmol) and Ph3CCl (11.4 g, 40.9 mmol) were added. The reaction mixture was stirred at room temperature for 2 h. The mixture was poured into water and extracted with DCM (30 mL×2). The organic layer was washed with brine, dried over anhydrous Na2SO4, and evaporated under vacuum. The residue was purified by flash chromatography on silica gel (PE:EA=10:1 to 5:1) to give (2S,4S)-4-hydroxy-1-tritylpyrrolidine-2-carboxylic acid methyl ester (7.60 g, two-step yield 48%) as a white solid. 1 H NMR (400MHz, CDCl3) δ7.60-7.50(m, 6H), 7.32-7.23(m, 6H), 7.21-7.13(m, 3H), 3.92-3.80(m, 3H), 3.65( s, 3H), 3.46 (d, J=11.5Hz, 1H), 2.80 (dd, J=11.5, 3.8Hz, 1H), 1.62 (d, J=13.8Hz, 1H), 1.38-1.11 (m, 1H).

[0383] Step 3. Methyl (2S,4S)-4-((methylsulfonyl)oxy)-1-tritylpyrrolidine-2-carboxylate

[0384] To a solution of (2S,4S)-4-hydroxy-1-tritylpyrrolidine-2-carboxylic acid methyl ester (10.0 g, 25.8 mmol) in DCM (100 mL) was added TEA (7.90 g, 78.0 mmol) and MsCl (4.50 g, 39.0 mmol). The reaction mixture was stirred at room temperature for 2 h. The mixture was diluted with water and extracted with DCM (80 mL×2). The organic layer was washed with brine, dried over anhydrous Na2SO4, and evaporated under vacuum to give a crude product of (2S,4S)-4-methanesulfonyloxy-1-tritylpyrrolidine-2-carboxylic acid methyl ester as a yellow oil, which was used directly in the next step.

[0385] Step 4. Methyl (2S,4R)-4-azido-1-tritylpyrrolidine-2-carboxylate

[0386] Sodium azide (7.80 g, 120 mmol) was added to a solution of (2S,4S)-4-methanesulfonyloxy-1-tritylpyrrolidine-2-carboxylic acid methyl ester in DMF (140 mL) in the above step. The reaction mixture was stirred at 80°C for 16 h, then diluted with water and extracted with EA (3×100 mL). The combined organic layers were washed with water, brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 10%) to give (2S,4R)-4-azido-1-tritylpyrrolidine-2-carboxylic acid methyl ester (8.00 g, two-step yield 64.5%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ7.58-7.51 (m, 6H), 7.33-7.24 (m, 6H), 7.18 (t, J=7.3Hz, 3H), 4.20-4.06 (m, 1H), 3.91 (d, J=8.9H z, 1H), 3.72 (dd, J=10.3, 7.6Hz, 1H), 3.63 (s, 3H), 2.63 (dd, J=10.3, 7.8Hz, 1H), 1.90-1.79 (m, 1H), 0.94-0.88 (m, 1H).

[0387] Step 5: ((2S,4R)-4-amino-1-tritylpyrrolidin-2-yl)methanol

[0388] To a stirred suspension of LiAlH4 (2.20 g, 58.0 mmol) in THF (80 mL) was added a solution of (2S, 4R)-4-azido-1-tritylpyrrolidine-2-carboxylic acid methyl ester (8.00 g, 19.4 mmol) in THF (10 mL) at 0°C. After the addition was complete, the reaction mixture was stirred at 0°C for 1 h and then quenched with water (2.5 mL), 15% NaOH solution (2.5 mL) and water (6 mL). After stirring for 0.5 h, the mixture was dried over Na2SO4, filtered and concentrated to give the crude product [(2S, 4R)-4-amino-1-trityl-pyrrolidin-2-yl]methanol (7.00 g) as a yellow oil, which was used in the next step without purification.

[0389] Step 6: tert-Butyl ((3R,5S)-5-(hydroxymethyl)-1-tritylpyrrolidin-3-yl)carbamate

[0390] To a solution of [(2S,4R)-4-amino-1-tritylpyrrolidin-2-yl]methanol (7.00 g, 19.5 mmol) in 1,4-dioxane (70 mL) was added TEA (3.90 g, 39.0 mmol) and (Boc)2O (5.10 g, 23.0 mmol). The mixture was stirred at room temperature for 2 h, then diluted with water and extracted with EA (3×100 mL). The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (PE:EA=20:1) to give tert-butyl N-[(3R,5S)-5-(hydroxymethyl)-1-tritylpyrrolidin-3-yl]carbamate (8.00 g, yield 89.3%) as a colorless oil. MS m / z 459.3 [M+H] + , 1 H NMR (400MHz, CDCl3) δ7.67-7.50(m, 6H), 7.33-7.23(m, 7H), 7.18(t, J=7.3Hz, 3H), 4.27-4.18(m, 1H), 3.77-3.64(m ,2H),3.58-3.51(m,1H),3.50-3.32(m,2H),2.60-2.48(m,1H),2.03-1.94(m,1H),1.90-1.81(m,1H),1.36(s,9H).

[0391] Step 7: tert-Butyl ((3R,5R)-5-fluoro-1-tritylpiperidin-3-yl)carbamate

[0392] DAST (2.80 mL, 21.0 mmol) was added dropwise to a stirred solution of tert-butyl N-[(3R, 5S)-5-(hydroxymethyl)-1-tritylpyrrolidin-3-yl]carbamate (6.80 g, 15.0 mmol) in THF (70 mL) at 0°C. The mixture was stirred at 0°C for 1 h, at room temperature for 1 h, and then cooled to 0°C again. A saturated aqueous solution of Na2CO3 was added to adjust the pH to 12. The phases were separated and the aqueous phase was extracted with EtOAc (2×40 mL). The organic extracts were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography (silica gel, PE / EtOAc=20 / 1) to give tert-butyl N-[(3R, 5R)-5-fluoro-1-trityl-3-piperidinyl]carbamate (3.90 g, 57% yield) as a white solid. MS m / z 483.3[M+Na] + .

[0393] Step 8: tert-Butyl ((3R,5R)-5-fluoropiperidin-3-yl)carbamate

[0394] AcOH (8 mL, 139.6 mmol) was added to a solution of tert-butyl N-[(3R,5R)-5-fluoro-1-trityl-3-piperidinyl]carbamate (3.90 g, 8.50 mmol) in MeOH (80 mL). The mixture was stirred at 80 °C for 2 h and then concentrated under reduced pressure. The residue was diluted with EA (20 mL) and H2O (20 mL). The mixture was acidified to pH = 2-3 with aqueous HCl solution and extracted with EA (10 mL×2). The aqueous layer was basified to pH = 9-10 with aqueous K2CO3 solution and extracted with DCM (50 mL×3). The DCM layers were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to give tert-butyl N-[(3R,5R)-5-fluoro-3-piperidinyl]carbamate (1.60 g, yield 87%) as a white solid. MS m / z 219.1[M+H] + , 1 H NMR (400MHz, DMSO-d6) δ6.61 (s, 1H), 4.77 (d, J=48.5Hz, 1H), 3.60 (br s, 1H), 2.96-2.82 (m, 2H), 2.77-2.62 (m, 1H), 2.43-2.33 (m, 1H), 2.09-1.97 (m, 1H), 1.78-1.55 (m, 1H), 1.39 (s, 9H).

[0395] Step 9: tert-Butyl ((3R,5R)-5-fluoro-1-methylpiperidin-3-yl)carbamate

[0396] To a solution of tert-butyl N-[(3R,5R)-5-fluoro-3-piperidinyl]carbamate (1.60 g, 7.30 mmol) in DCE (20 mL) was added formic acid (1.20 g, 26.0 mmol) and (HCHO)n (0.75 g, 25 mmol), and stirred at room temperature for 1 h. NaBH(OAc)3 (5.4 g, 25.0 mmol) was added to the mixture, and stirred at room temperature for 16 h. The reaction mixture was quenched with water, basified with aqueous K2CO3 solution, and extracted with DCM. The organic phases were combined and washed with brine. The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by silica gel chromatography (PE / EA=3 / 1) to give tert-butyl N-[(3R,5R)-5-fluoro-1-methyl-3-piperidinyl]carbamate (1.00 g, yield 59.0%) as a white solid. MS m / z 233.1[M+H] + .

[0397] Step 10. (3R,5R)-5-Fluoro-1-methylpiperidin-3-amine hydrochloride

[0398] To a solution of tert-butyl N-[(3R,5R)-5-fluoro-1-methyl-3-piperidinyl]carbamate (1.00 g, 4.30 mmol) in EA (10 mL) was added a solution of hydrochloric acid in 1,4-dioxane (14 mL, 56 mmol, 4 mol / L). The resulting mixture was stirred at room temperature for 2 h, then filtered, and the solid was washed with EA (2 mL) and dried in vacuo to give a white solid (3R,5R)-5-fluoro-1-methylpiperidin-3-amine hydrochloric acid (750 mg, yield 75%). MS m / z 133.1 [M+H] + , 1 H NMR (400MHz, DMSO-d6) δ11.18 (d, J=3.6Hz, 1H), 8.94 (s, 3H), 5.25 (d, J=44.4Hz, 1H), 3.72-3.66 (m, 2H), 3.56 -3.50 (m, 1H), 3.41-3.27 (m, 1H), 3.13 (t, J=11.2Hz, 1H), 2.85 (s, 3H), 2.47-2.41 (m, 1H), 2.01-1.83 (m, 1H).

[0399] Intermediate 3g: (3R,5R)-5-fluoro-1-ethylpiperidin-3-amine hydrochloride

[0400]

[0401] The starting material, tert-butyl ((3R,5R)-5-fluoropiperidin-3-yl)carbamate, was prepared according to steps 1 to 8 of intermediate 3f.

[0402] The title compound was prepared in analogy to the procedure for Intermediate 3b, using tert-butyl ((3R,5R)-5-fluoropiperidin-3-yl)carbamate in place of tert-butyl (R)-piperidin-3-ylcarbamate in step 1. MS m / z 147.2 [M+H] + ; 1 HNMR (400MHz, DMSO-d6) δ11.02 (s, 1H), 8.94 (s, 3H), 5.27 (d, J=44..8Hz, 1H), 3.74-3.62 (m, 3H), 3.32 -3.22 (m, 3H), 3.09-3.05 (m, 1H), 2.48-2.43 (m,, 1H), 2.06-1.89 (m, 1H), 1.28-1.25 (t, J=14.0Hz, 3H).

[0403] Example 1

[0404] Preparation of Compound 1 (1s,3s)-3-((1-(4-methoxy-2-(2,2,2-trifluoroethyl)phenyl)pyrrolo[1,2-d][1,2,4]triazin-4-yl)amino)-1-methylcyclobutane-1-ol

[0405]

[0406] Step 1 :1-[4-methoxy-2-(2,2,2-trifluoroethyl)phenyl]-4-methanesulfonyl-pyrrolo[1,2-d][1,2,4]triazine

[0407] 1-Chloro-4-methanesulfonyl-pyrrolo[1,2-d][1,2,4]triazine (Intermediate 1c, 0.380 g, 1.90 mmol), 2-[4-methoxy-2-(2,2,2-trifluoroethyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Intermediate 2a, 0.842 g, 2.66 mmol) and XPhos Pd G3 (0.164 g, 0.190 mmol) were added to a vial, evacuated and refilled with argon, and then dioxane (9.5 mL) and K2CO3 (2M, 2.9 mL, 5.71 mmol) were added. The mixture was sparged with argon for 5 min and then heated to 95 °C for 3 h. After cooling to room temperature, the mixture was diluted with EtOAc and filtered through Celite. The filtrate was washed with brine, dried (MgSO4), filtered and concentrated. Purification by chromatography on SiO2 (EtOAc:hexanes, 0-30%) afforded a pink solid (0.329 g, 49%). MS m / z 354.4 [M+H] + .

[0408] Step 2 :(1s,3s)-3-((1-(4-methoxy-2-(2,2,2-trifluoroethyl)phenyl)pyrrolo[1,2-d][1,2,4]triazin-4-yl)amino)-1-methylcyclobutane-1-ol

[0409] A solution of 1-[4-methoxy-2-(2,2,2-trifluoroethyl)phenyl]-4-methanesulfonyl-pyrrolo[1,2-d][1,2,4]triazine (0.070 g, 0.18 mmol) and 3-amino-1-methylcyclobutanol hydrochloride (0.030 g, 0.22 mmol) in DMA (0.10 mL) and iPr2Net (0.10 mL, 0.57 mmol) was heated to 135°C for 4 h. The reaction was cooled to room temperature and diluted with DCM / iPrOH (9:1). The solution was washed with brine, dried (Na2SO4), filtered and concentrated. Chromatographic purification on SiO2 (MeOH:DCM, 0-10%) followed by reverse phase chromatography (0.1% formic acid in MeCN: 0.1% formic acid in water, 5-100%) afforded a white solid (0.010 g, 28%). MS m / z 407.3[M+H] + ; 1H NMR (500MHz, methanol-d4) δ 7.83 (d, J = 1.8 Hz, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.10 (s, 1H), 7.09-7.04 (m, 1H), 6.96 (t, J = 3.4 Hz, 1H), 6.53 (d, J = 3.8 Hz, 1H), 4.31-4.21 (m, 1H), 3.89 (s, 3H), 3.69 (q, J = 11.14 Hz, 2H), 2.69-2.62 (m, 2H), 2.31-2.22 (m, 2H), 1.44 (s, 3H). 2H (NH and OH) were not observed.

[0410] The following compounds were prepared according to the procedure of Example 1 by substituting appropriate starting materials, reagents and reaction conditions.

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430]

[0431] Example 2

[0432] Preparation of Compound 2 (R)-N-(1-methylpiperidin-3-yl)-1-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)imidazo[1,5-d][1,2,4]triazine-4-yl

[0433] amine

[0434]

[0435] Step 1. 3-(Methylthio)-6-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,2,4-triazine

[0436] To a solution of 4,4,5,5-tetramethyl-2-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,3,2-dioxaborolane (Intermediate 2b, 2 g, 5.7 mmol, 1.0 eq.) in 1,4-dioxane / H2O (5:1, 24 mL) was added 6-bromo-3-methylthio-1,2,4-triazine (1.1 g, 5.1 mmol, 1 eq.), Pd(dppf)Cl2 (372.6 mg, 0.1 eq.) and K3PO4 (2.16 g, 2 eq.) under N2 atmosphere at room temperature. The reaction mixture was stirred at 90°C under N2 for 2 h, then diluted with water (20 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 10%) to give 3-(methylthio)-6-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,2,4-triazine (1.6 g, yield 81.1%) as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ 8.95 (s, 1H), 8.03-7.97 (m, 2H), 7.89 (d, J=8.0Hz, 1H), 4.10 (q, J=11.2Hz, 2H), 2.70 (s, 3H).

[0437] Step 2. 4-(Methylthio)-1-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)imidazo[1,5-d][1,2,4]triazine

[0438] To a solution of TosMIC (1.1 g, 5.4 mmol, 1.2 equiv) and DBU (827.4 mg, 5.4 mmol, 1.2 equiv) in DCE (20 mL) was added 3-(methylthio)-6-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-1,2,4-triazine (1.6 g, 4.5 mmol, 1.0 equiv) under N2 at room temperature. The reaction mixture was stirred at room temperature for 2 h, then diluted with water (50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by flash chromatography on silica gel (PE:EA=0% to 25%) to give the title product (1.29 g, 44.9% yield) as a yellow oil. MS m / z 392.9[M+1] + ; 1H NMR (400MHz, DMSO-d6) δ 8.84 (s, 1H), 8.04 (s, 1H), 7.95 (dd, J=19.0, 8.2Hz, 2H), 7.71 (s, 1H), 4.13-4.04 (m, 2H), 2.89 (s, 3H).

[0439] Step 3. (R)-N-(1-methylpiperidin-3-yl)-1-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)imidazo[1,5-d][1,2,4]triazin-4-amine

[0440] To a solution of 4-(methylthio)-1-(2-(2,2,2-trifluoroethyl)-4-(trifluoromethyl)phenyl)-imidazo[1,5-d][1,2,4]triazine (100 mg, 0.25 mmol, 1.0 equiv) in DMA (1 mL) was slowly added (R)-1-methylpiperidin-3-amine dihydrochloride (119 mg, 0.64 mmol, 2.5 equiv) and DIEA (329.4 mg, 10.0 equiv) under N2 at room temperature. The reaction mixture was stirred at 140 °C under N2 for 12 h, then filtered and concentrated under reduced pressure. The crude product was purified by reverse phase chromatography (0.1% formic acid in MeCN: 0.1% formic acid in water, 5-100%) to afford the title product (16.4 mg, 13.8% yield) as a pale yellow solid formate salt. MS m / z 459.0 [M+1] + ; 1 H NMR (400MHz, methanol-d4) δ8.77 (s, 1H), 8.49 (s, 1H), 7.92 (s, 1H), 7.85 (dd, J=19.2, 8.2Hz, 2H), 7.50 (s, 1H), 4.63-4.36 (m, 1H), 3.92 (q, J =10.9Hz, 2H), 3.64-3.47(m, 1H), 3.29-3.08(m, 2H), 3.02-2.79(m, 2H), 2.72-2.57(m, 1H), 2.31-1.70(m, 4H), 1.28(t, J=6.8Hz, 3H).

[0441] The following compounds were prepared according to the procedure of Example 2 by substituting appropriate starting materials, reagents and reaction conditions.

[0442]

[0443]

[0444]

[0445]

[0446]

[0447]

[0448]

[0449]

[0450]

[0451]

[0452]

[0453]

[0454]

[0455]

[0456]

[0457]

[0458]

[0459]

[0460] Example 3

[0461] Preparation of Compound 3 (1s,3s)-1-methyl-3-((2-methyl-4-(2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7-yl)

[0462] Amino)cyclobutane-1-ol

[0463]

[0464] Step 1 .2-Methyl-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine-4-ol

[0465] A cold solution of 7-bromo-2-methyl-pyrazolo[1,5-d][1,2,4]triazine-4-ol (Intermediate 1f, 600.0 mg, 2.620 mmol) in dry DMF (2.6 mL) was treated with sodium thiomethoxide (386.6 mg, 5.239 mmol) at 0°C. The mixture was then stirred at room temperature for 30 minutes. Upon completion, DMF was evaporated and the resulting oily residue was precipitated with water. The mixture was then cooled at 0°C and the precipitate was filtered while cold. The collected precipitate was washed with water and dried overnight to give 2-methyl-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine-4-ol (476.0 mg, 93% yield) as a white solid. MS m / z 197.1[M+H] + .

[0466] Step 2. 4-Chloro-2-methyl-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine

[0467] A mixture of 2-methyl-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine-4-ol (476.0 mg, 2.426 mmol) and phosphorus oxychloride (7.44 g, 4.51 mL, 48.51 mmol) was heated at 75°C for 6 hours and the solution became clear. The excess POCl3 was then evaporated and the resulting residue was precipitated with water. The pH of the aqueous solution was adjusted to neutral (pH ~ 7) using saturated NaHCO3 solution. The precipitate was filtered, washed with water, and dried overnight to give 4-chloro-2-methyl-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine (383 mg, 74% yield) as a white solid. MS m / z 215.1[M+H] + .

[0468] Step 3. 2-Methyl-7-(methylthio)-4-(2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl)-pyrazolo[1,5-d][1,2,4]triazine

[0469] A mixture of 4-chloro-2-methyl-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine (38.8 mg, 0.181 mmol), [2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl]boronic acid (64.4 mg, 0.235 mmol), XPhos Pd G4 (16.4 mg, 0.0181 mmol) and potassium carbonate (74.9 mg, 0.542 mmol) was dissolved in dioxane (2.1 mL) and water (0.4 mL). The reaction vessel was evacuated and backflushed with nitrogen (3×), and the mixture was stirred at 80°C under nitrogen for 4 hours. After the reaction was complete, the mixture was diluted with EtOAc, washed with water and brine, dried over Na2SO4, and then evaporated under vacuum. The crude product was purified by flash column chromatography using 0-80% EtOAc in hexane to give 2-methyl-7-(methylthio)-4-(2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl)-pyrazolo[1,5-d][1,2,4]triazine (25.5 mg, 35% yield) as a brown solid. MS m / z 409.1 [M+H] + .

[0470] Step 4. (1s,3s)-1-Methyl-3-((2-methyl-4-(2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazin-7-yl)amino)cyclobutane-1-ol

[0471] To a solution of 2-ethyl-7-(methylthio)-4-(2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine (25.5 mg, 0.0625 mmol) in NMP (0.50 mL) was added (1s,3s)-3-amino-1-methylcyclobutane-1-ol hydrochloride (25.8 mg, 0.187 mmol) and N,N-diisopropylethylamine (48.4 mg, 65.4 μL, 0.375 mmol). The mixture was heated at 130°C under nitrogen for 24 hours to achieve partial conversion. Additional (1s,3s)-3-amino-1-methylcyclobutane-1-ol hydrochloride (25.8 mg, 0.187 mmol) was then added and the reaction was further stirred and heated at 140°C for 24 hours. After complete conversion to product, the solution was diluted with a small amount of water (0.50 mL). The black solution was purified by reverse phase chromatography (0.1% formic acid in MeCN: 0.1% formic acid in water, 5-100%) to give a white solid (1s, 3s)-1-methyl-3-((2-methyl-4-(2-(trifluoromethoxy)-4-(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazin-7-yl)amino)cyclobutane-1-ol (18.2 mg, 63% yield). MS m / z 462.1[M+H] + ; 1 H NMR (500 MHz, methanol-d4) δ 7.95 (d, J = 8.1 Hz, 1H), 7.89 (d, J = 8.1 Hz, 1H), 7.81 (s, 1H), 6.51 (s, 1H), 4.39-4.31 (m, 1H), 2.71-2.61 (m, 2H), 2.54 (s, 3H), 2.36-2.26 (m, 2H), 1.44 (s, 3H).

[0472] The following compounds were prepared according to the procedure of Example 3 by substituting appropriate starting materials, reagents and reaction conditions.

[0473]

[0474]

[0475]

[0476]

[0477]

[0478] Example 4

[0479] Preparation of Compound 4 (1s,3s)-3-((4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazin-7-yl)amino)-1-methylcyclobutane-1-ol

[0480]

[0481] Step 1: (2,4-bis(trifluoromethyl)phenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methanol

[0482] At -78 ° C, n-butyl lithium (16.8 mL, 1.2 eq., 2.5 M hexane solution) was added dropwise to a solution of 1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole (5.3 g, 35.0 mmol, 1.0 eq.) in tetrahydrofuran (170 mL). The reaction mixture was stirred at -78 ° C for 30 min, and then 2,4-bis(trifluoromethyl)benzaldehyde (11.02 g, 1.3 eq.) was added. The mixture was then warmed to room temperature and stirred for 2 hours, and then quenched with water (200 mL). The resulting mixture was extracted with ethyl acetate (3×100 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure to give crude (2,4-bis(trifluoromethyl)phenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methanol (15.0 g, crude) as a dark yellow oil, which was used in the next step without further purification.

[0483] Step 2: (2,4-bis(trifluoromethyl)phenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methanone

[0484] To a solution of (2,4-bis(trifluoromethyl)phenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methanol (15.0 g, crude) in dichloromethane (170 mL) was added Dess-Martin periodinane (19.3 g, mmol, 1.3 equiv) portionwise at 0°C. The reaction mixture was stirred at room temperature for 2 h and then washed with saturated Na2S2O3 solution (2×100 mL), saturated NaHCO3 solution (100 mL) and brine (100 mL). The organic phase was dried over sodium sulfate, filtered and concentrated under reduced pressure to give (2,4-bis(trifluoromethyl)phenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methanone (15 g, crude) as a dark yellow oil which was used in the next step without further purification. 1H NMR (400MHz, DMSO-d6) δ8.26 (s, 1H), 8.23 ​​(d, J = 8.4Hz, 1H), 7.93 (d, J = 8.0Hz, 1H), 7.68 (d, J = 2.0Hz, 1H),, 6.66 (d, J = 2.0Hz, 1H), 6.17 (dd, J=10.0, 2.0Hz, 1H), 3.95 (d, J=11.6Hz, 1H), 3.71-3.57 (m, 1H), 2.38-2.27 (m, 1H), 2.07-1.93 (m, 2H), 1.78-1.64 (m, 1H), 1.61-1.51 (m, 2H).

[0485] Step 3. (2,4-Bis(trifluoromethyl)phenyl)(1H-pyrazol-3-yl)methanone

[0486] (2,4-bis(trifluoromethyl)phenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methanone (15 g, crude product) was dissolved in a solution of dichloromethane (300 mL) and trifluoroacetic acid (30 mL). The resulting mixture was stirred at room temperature for 16 h. After the reaction was completed, saturated NaHCO3 solution was added to adjust the pH of the reaction mixture to 7. The organic phase was separated, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography and eluted with 0-50% EtOAc hexane solution to give (2,4-bis(trifluoromethyl)phenyl)(1H-pyrazol-3-yl)methanone (6.50 g, 3-step yield 60.2%) as a yellow solid. MS m / z 309.0 [M+H] + ; 1 HNMR (400MHz, DMSO-d6) δ13.68 (s, 1H), 8.21 (s, 1H), 8.18 (d, J=8.4Hz, 1H), 7.99 (s, 1H), 7.88 (d, J=8.0Hz, 1H), 6.98 (s, 1H).

[0487] Step 4. 4-(2,4-Bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7(6H)-one

[0488] A mixture of (2,4-bis(trifluoromethyl)phenyl)(1H-pyrazol-3-yl)methanone (3.0 g, 9.7 mmol, 1.0 equiv) and ethyl carbazate (1.01 g, 1.0 equiv) in mesitylene (100 mL) was heated at 165°C for 4 h. After the reaction was complete, the reaction mixture was cooled to room temperature and then directly purified by silica gel column chromatography, eluting with 0-40% EtOAc hexane solution to give a pale yellow solid 4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7(6H)-one (1.50 g, 44.4% yield). MS m / z 349.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ13.68 (s, 1H), 8.21 (s, 1H), 8.18 (d, J=8.0Hz, 1H), 7.99 (dd, J=2.8, 1.6Hz, 1H), 7.88 (d, J=8.0Hz, 1H), 6.98 (dd, J=2.4, 2.0Hz, 1H).

[0489] Step 5. 4-(2,4-Bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7(6H)-thione

[0490] A mixture of 4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7(6H)-one (1.50 g, 4.31 mmol, 1.0 equiv) and Lawesson's reagent (3.49 g, 2.0 equiv) in toluene (40 mL) was heated at 120 °C for 16 h. After the reaction was complete, the reaction mixture was cooled to room temperature and then filtered through a Celite pad. The filtrate was washed with water (80 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure to give 4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7(6H)-thione (1.30 g, crude) as a pale yellow solid, which was used in the next step without further purification.

[0491] Step 6. 4-(2,4-bis(trifluoromethyl)phenyl)-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine

[0492] To a mixture of 4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazine-7(6H)-thione (1.30 g, 1.0 eq.) and K2CO3 (1.49 g, 2.5 eq.) in THF (12 mL) and water (6 mL) was added iodomethane (0.4 mL, 1.5 eq.) dropwise. The mixture was stirred at room temperature for 1 hour, then diluted with water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with 0-30% EtOAc in hexanes to give 4-(2,4-bis(trifluoromethyl)phenyl)-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine (1.08 g, 66.2% yield over 2 steps) as a pale yellow solid. MS m / z 379.3[M+H] + .

[0493] Step 7. (1s,3s)-3-((4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazin-7-yl)amino)-1-methylcyclobutane-1-ol

[0494] A mixture of 4-(2,4-bis(trifluoromethyl)phenyl)-7-(methylthio)pyrazolo[1,5-d][1,2,4]triazine (200 mg, 0.53 mmol, 1.0 equiv), (1s,3s)-3-amino-1-methylcyclobutane-1-ol hydrochloride (180.9 mg, 2.5 equiv) and DIEA (1.20 mL, 13.0 equiv) in DMAc (0.75 mL) was heated at 140 °C for 4 h. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with water (50 mL), and extracted with EtOAc (3×50 mL). The organic phase was washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography to give (1s,3s)-3-((4-(2,4-bis(trifluoromethyl)phenyl)pyrazolo[1,5-d][1,2,4]triazin-7-yl)amino)-1-methylcyclobutane-1-ol (68.9 mg, yield 30.1%) as a white solid. MS m / z 432.1 [M+H] + . 1 H NMR (400 MHz, methanol-d4) δ 8.21-8.18 (m, 2H), 8.14 (d, J = 8.4 Hz, 1H), 7.88 (d, J = 8.0 Hz, 1H), 6.56 (d, J = 2.0 Hz, 1H), 4.36 (quin, J = 8.0 Hz, 1H), 2.71-2.62 (m, 2H), 2.37-2.29 (m, 2H), 1.44 (s, 3H).

[0495] The following compounds were prepared according to the procedure of Example 4 by substituting appropriate starting materials, reagents and reaction conditions.

[0496]

[0497] Biological detection

[0498] IL-1β secretion detection:

[0499] According to the supplier's instructions, monocytic THP-1 cells (ATCC: TIB-202) were cultured in a growth medium consisting of RPMI 1640 medium (ThermoFisher, Cat#11875-085), 10% FBS (ThermoFisher) and 0.05 mM β-mercaptoethanol (ThermoFisher, Cat#21985-023). The cell concentration was adjusted to 7.5 × 10 5 Cells / mL were cultured in complete growth medium containing a final concentration of 100ng / mL phorbol-12-myristate-13-acetate (PMA, Sigma#P8139). Cells were seeded into 96-well cell culture plates (ThermoFisher, Cat#165305) at 100μL / well and differentiated for 24h in a cell culture incubator at 37°C and 5% CO2. The cells were rinsed once with 100μl PBS and then replaced with fresh RPMI+5% FBS. The compounds were diluted 3-fold with DMSO to obtain a total of 7 concentrations. The diluted compounds were added to the cells at a ratio of 1:200 and incubated for 20h. 2.5μM Nigericin (Sigma: Cat#SML1779-1ml) was added to activate the NLRP3 inflammasome for 3h. After the incubation, 15 μL of conditioned medium was removed and the IL-1β level was detected using the HTRF IL-1β detection kit (Cisbio: Cat#62HIL1BPEH) according to the manufacturer's instructions.

[0500] The compounds produced and prepared according to the present invention can be evaluated by various tests to characterize their activity. For example, NLRP3-dependent IL 1β secretion was evaluated in THP1 cells. The IC50 value for inhibiting IL 1β was calculated by logistic fitting through a plot of the percentage of inhibition versus the concentration of the inhibitor. Table I describes examples of compounds according to Formula I. Data <1nM are represented as *****; data 1-10nM are represented as ****; data 10-100nM are represented as ***, data 100-300nM are represented as **, and data ≥300nM are represented as *. The data obtained from the THP1 NLRP3-dependent IL-1β secretion assay indicate that the compounds of the present invention can be used to treat diseases mediated by NLRP3 activation.

[0501] Table I

[0502]

[0503]

[0504]

[0505]

[0506] All documents cited herein are incorporated by reference into this application for any and all purposes to the same extent as if each individual reference was fully set forth herein, regardless of whether or not such document is specifically and individually indicated as being incorporated by reference.

[0507] Although certain embodiments have been described in detail above, those skilled in the art will readily appreciate that many modifications may be made to the embodiments without departing from the teachings thereof. All such modifications are intended to be included within the scope of the claims presented herein.

Claims

1. A compound selected from: in, The form of the compound is selected from the group consisting of pharmaceutically acceptable salts, hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers, tautomers, and isotopically enriched forms thereof.

2. A compound selected from: in, The form of the compound is selected from the group consisting of pharmaceutically acceptable salts, hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers, tautomers, and isotopically enriched forms thereof.

3. A compound selected from: in, The form of the compound is selected from the group consisting of pharmaceutically acceptable salts, hydrates, solvates, racemates, enantiomers, diastereomers, stereoisomers, tautomers, free bases and isotopically enriched forms thereof.

4. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

5. A method for treating or ameliorating a disease regulated by NLRP3 in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 3.

6. The method of claim 5 for treating or ameliorating a disease regulated by NLRP3, wherein the disease is selected from the group consisting of Alzheimer's disease, frontotemporal dementia (FTD), Huntington's disease, Parkinson's disease, perioperative neurocognitive disorder, cognitive disorder after cardiac arrest, cognitive disorder after stroke, sepsis, sepsis-related encephalopathy, subarachnoid hemorrhage, macular degeneration, retinal neovascularization, uveitis, colitis, endothelial dysfunction, gout, pseudogout, graft-versus-host disease (GvHD), systemic lupus erythematosus-lupus nephritis, cryopyrin-associated periodic syndrome (CAPS), cystic fibrosis, sickle cell disease, VCP Related diseases, liver fibrosis, non-alcoholic fatty liver disease (NASH), muscle atrophy, inherited and acquired myopathies such as Duchenne muscular dystrophy (DMD), hyperalgesia, multiple sclerosis-related neuropathic pain, acute kidney injury, chronic crystalline nephropathy, chronic kidney disease, asthma and allergic airway inflammation, diabetes-related atherosclerosis, diabetic encephalopathy, diabetic nephropathy, islet transplant rejection, obesity-related nephropathy, oxalate nephropathy, renal fibrosis, renal hypertension, type I diabetes, type II diabetes, psoriasis, hidradenitis suppurativa, atherosclerosis, and cytokine release syndrome (CRS).

7. The method according to any one of claims 5 to 6, wherein the effective amount of the compound ranges from about 0.001 mg / kg / day to about 500 mg / kg / day.

8. A compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof for use in treating or ameliorating a disease regulated by NLRP3, the disease being selected from Alzheimer's disease, frontotemporal dementia (FTD), Huntington's disease, Parkinson's disease, perioperative neurocognitive disorder, cognitive disorder after cardiac arrest, cognitive disorder after stroke, sepsis, sepsis-related encephalopathy, subarachnoid hemorrhage, macular degeneration, retinal neovascularization, uveitis, colitis, endothelial dysfunction, gout, pseudogout, graft-versus-host disease (GvHD), systemic lupus erythematosus-lupus nephritis, cryopyrin-associated periodic syndrome (CAPS ), cystic fibrosis, sickle cell disease, VCP-related diseases, liver fibrosis, nonalcoholic fatty liver disease (NASH), muscular dystrophy, inherited and acquired myopathies, hyperalgesia, neuropathic pain in multiple sclerosis, acute kidney injury, chronic crystalline nephropathy, chronic kidney disease, asthma and allergic airway inflammation, diabetes-related atherosclerosis, diabetic encephalopathy, diabetic nephropathy, islet transplant rejection, obesity-related kidney disease, oxalate nephropathy, renal fibrosis, renal hypertension, type I diabetes, type II diabetes, psoriasis, hidradenitis suppurativa, atherosclerosis, and cytokine release syndrome (CRS).

9. The use of a compound according to claim 8, wherein the effective amount of the compound ranges from about 0.001 mg / kg / day to about 500 mg / kg / day.

10. Use of a compound according to any one of claims 1 to 3 in the preparation of a pharmaceutical composition for treating or ameliorating a disease regulated by NLRP3 in a subject in need thereof, comprising administering to the subject an effective amount of the compound or a form thereof mixed with one or more pharmaceutically acceptable excipients.

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