Novel compounds as modulators of NLRP3 inhibition

Abstract

The present invention relates to novel compounds having the general formula (I) # imgabs0 # wherein R1, R2, R3, R4, R5 and R6 are as described herein; comprising the compound; and methods of using the compounds.

Description

Technical Field

[0001] The present invention relates to organic compounds useful for the treatment and / or prevention in mammals, and in particular to compounds that modulate NLRP3 inhibition.

[0002] The present invention provides novel compounds of formula I,

[0003]

[0004] wherein

[0005] R 1 and R 5 together with the atoms to which they are attached form

[0006] i. a 4- to 6-membered heterocyclic ring containing a single O heteroatom, or

[0007] ii. a 4- to 5-membered cycloalkyl ring;

[0008] R 2 and R 3 are independently selected from H and alkyl, provided that only one of R 2 or R 3 can be H or alkyl;

[0009] R 4 is selected from

[0010] i. alkyl,

[0011] ii. hydroxyalkyl,

[0012] iii. alkoxyalkyl,

[0013] iv. cycloalkyl substituted with hydroxy or alkoxy,

[0014] v. cycloalkylalkyl substituted with hydroxy, alkoxy or –COOH,

[0015] vi. a 4- to 6-membered heterocyclic ring containing a single O heteroatom,

[0016] vii. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 2 or 3,

[0017] viii. heteroarylalkyl, where the heteroaryl in the heteroarylalkyl is a tetrazole, oxadiazole or oxazole heteroaryl,

[0018] ix. heteroarylalkyl, where the heteroaryl in the heteroarylalkyl is pyrazole,

[0019] x. -(CH2)n S(O)2CH3 or (CH2) n CN, where n is 3,

[0020] xi.-(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3, R'

[0021] is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form

[0022] a. a 5-membered heterocycle, where the heterocycle is optionally substituted with OH; or

[0023] b. a 6-membered heterocycle additionally containing 1 O heteroatom;

[0024] R 6 is H or –OH;

[0025] where when R 6 is –OH, R 4 can only be alkyl;

[0026] and pharmaceutically acceptable salts.

[0027] In addition, the present invention includes all racemic mixtures, all their corresponding enantiomers and / or optical isomers. Background Art

[0028] The NOD-like receptor (NLR) family (the NLRP3 inflammasome, a protein containing a pyrin domain) is a component of the inflammatory process, and its abnormal activity is pathogenic in genetic disorders such as cryopyrin-associated periodic syndromes (CAPS) and complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer's disease, and atherosclerosis.

[0029] NLRP3 is an intracellular signaling molecule that can sense many pathogen-derived, environmental, and host-derived factors. Upon activation, NLRP3 binds to apoptosis-associated speck-like protein, which contains a caspase activation and recruitment domain (ASC). Then, ASC polymerizes to form large aggregates called ASC specks. The polymeric ASC then interacts with the cysteine protease caspase-1 to form a complex called the inflammasome. This leads to the activation of caspase-1, which cleaves the precursor forms of the pro-inflammatory cytokines IL-1β and IL-18 (called pro-IL-1β and pro-IL-18, respectively), thus activating these cytokines. Caspase-1 also mediates a type of inflammatory cell death called pyroptosis. ASC specks can also recruit and activate caspase-8, which can process pro-IL-1β and pro-IL-18 and trigger apoptotic cell death.

[0030] Caspase-1 cleaves pro-IL-1β and pro-IL-18 into their active forms, which are secreted by cells. Active Caspase-1 also cleaves gasdermin-D to trigger pyroptosis. Caspase-1 can also mediate the release of alarm protein molecules such as IL-33 and high mobility group box 1 (HMGB1) by controlling the pyroptotic cell death pathway. Caspase-1 also cleaves intracellular IL-1R2, leading to its degradation and the release of IL-1α. In human cells, Caspase-1 can also control the processing and secretion of IL-37. Many other substrates of Caspase-1, such as components of the cytoskeleton and glycolytic pathways, may contribute to Caspase-1-dependent inflammation.

[0031] NLRP3-dependent ASC specks are released into the extracellular environment, where the specks can activate Caspase-1, induce the processing of Caspase-1 substrates, and propagate inflammation.

[0032] The active cytokines derived from NLRP3 inflammasome activation are important drivers of inflammation and interact with other cytokine pathways to form an immune response to infection and injury. For example, IL-1β signaling induces the secretion of the pro-inflammatory cytokines IL-6 and TNF. IL-1β and IL-18 cooperate with IL-23 to induce memory CD4 Th17 cells and γδ T cells to produce IL-17 in the absence of T cell receptor engagement. IL-18 and IL-12 also cooperate to induce IFN-γ production from memory T cells and NK cells, driving a Th1 response.

[0033] The inherited CAPS diseases Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), and neonatal-onset multisystem inflammatory disease (NOMID) are caused by gain-of-function mutations in NLRP3, thus defining NLRP3 as a key component of the inflammatory process. NLRP3 is also associated with the pathogenesis of many complex diseases, which particularly include metabolic disorders such as type 2 diabetes, atherosclerosis, obesity, and gout.

[0034] The role of NLRP3 in central nervous system diseases is emerging, and lung diseases have also been shown to be affected by NLRP3. NLRP3 has also been implicated in many central nervous system conditions, including Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, and brain injury caused by pneumococcal meningitis (Walsh et al., Nature Reviews, 15:84-97, 2014, and Dempsey et al. Brain. Behav. Immun. 2017 61:306-316). NLRP3 has also been shown to play a role in many lung diseases, including chronic obstructive pulmonary disease (COPD), asthma (including steroid-resistant asthma), asbestosis, and silicosis (DeNardo et al., Am. J. Pathol., 184:42-54, 2014 and Kim et al. Am J Respir Crit Care Med. 2017 196(3):283-97). In addition, NLRP3 has a role in the development of liver disease, kidney disease, and aging. Many of these associations have been defined using Nlrp3 - / - mice, but there are also insights into the specific activation of NLRP3 in these diseases. In type 2 diabetes (T2D), the deposition of islet amyloid polypeptide in the pancreas activates NLRP3 and IL-1β signaling, leading to cell death and inflammation.

[0035] Several small molecules have been shown to inhibit the NLRP3 inflammasome. Glibenclamide inhibits IL-1β production at micromolar concentrations in response to the activation of NLRP3 but not NLRC4 or NLRP1. Other previously characterized weak NLRP3 inhibitors include parthenolide, 3,4-methylenedioxy-β-nitrostyrene, and dimethyl sulfoxide (DMSO), although these agents have limited potency and are non-specific.

[0036] Current treatments for NLRP3-related diseases include biologic agents that target IL-1. These are the recombinant IL-1 receptor antagonist anakinra, the neutralizing IL-1β antibody canakinumab, and the soluble decoy IL-1 receptor rilonacept. These approaches have been shown to successfully treat CAPS, and these biologic agents have been used in clinical trials for other IL-1β-related diseases.

[0037] There is a need to provide compounds having improved pharmacological and / or physiological and / or physicochemical properties, and / or compounds that provide useful alternatives to known compounds. Further, when developing NLRP3 inhibitors for treating peripheral indications, it is advantageous to minimize the exposure of the NLRP3 inhibitory compound in the brain relative to systemic exposure to minimize the risk of potential side effects in the central nervous system (CNS). The compounds of formula I achieve this by showing increased efflux and / or decreased permeability in a transcellular assay expressing active P-gp protein. P-gp (P-glycoprotein) is an important transport protein that is expressed in capillary endothelial cells that form the blood-brain barrier and the blood-testis barrier (where it pumps xenobiotics back into the capillaries) and limits brain exposure. SUMMARY OF THE INVENTION

[0038] The present invention provides novel compounds of formula I,

[0039]

[0040] wherein

[0041] R 1 and R 5 together with the atoms to which they are attached form

[0042] i. a 4- to 6-membered heterocyclic ring containing a single O heteroatom, or

[0043] ii. a 4- to 5-membered cycloalkyl ring;

[0044] R 2 and R 3 are selected from H and alkyl, where only one of R 2 or R 3 can be H or alkyl;

[0045] R 4 is selected from

[0046] i. alkyl,

[0047] ii. hydroxyalkyl,

[0048] iii. alkoxyalkyl,

[0049] iv. cycloalkyl substituted with hydroxy or alkoxy,

[0050] v. cycloalkylalkyl substituted with hydroxy, alkoxy or –COOH,

[0051] vi. a 4- to 6-membered heterocyclic ring containing a single O heteroatom,

[0052] vii. (CH2) n C(O)OH, (CH2) nC(O)OCH3 or (CH2) n C(O)NHCH3, where n is 2 or 3,

[0053] viii. heteroarylalkyl, where the heteroaryl in the heteroarylalkyl is a tetrazole, oxadiazole or oxazole heteroaryl,

[0054] ix. heteroarylalkyl, where the heteroaryl in the heteroarylalkyl is pyrazole;

[0055] x. -(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3,

[0056] xi. -(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3,

[0057] R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form

[0058] a. a 5-membered heterocycle, where the heterocycle is optionally substituted with OH; or

[0059] b. a 6-membered heterocycle additionally containing 1 O heteroatom;

[0060] R 6 is H or –OH,

[0061] where when R 6 is –OH, R 4 can only be alkyl;

[0062] and pharmaceutically acceptable salts.

[0063] The term "alkyl" refers to a monovalent straight-chain or branched-chain saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, unless otherwise specified, alkyl contains 1 to 6 carbon atoms (C 1-6 -alkyl) or 1 to 4 carbon atoms (C 1-4 -alkyl). Examples of C 1-6 -alkyl include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and pentyl. Particular alkyls include methyl and ethyl. Other specific alkyl groups are propyl.

[0064] The term "alkoxy" refers to a group of the formula -O-R', where R' is a C 1-6 -alkyl group. Examples of C 1-6 -alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy.

[0065] The term "alkoxyalkyl" means an alkyl group in which one of the hydrogen atoms of the alkyl group has been replaced by an alkoxy group. Examples of alkoxyalkyl are methoxymethyl and methoxyethyl. Specific examples of alkoxyalkyl are methoxyethyl and methoxypropyl.

[0066] The term "cycloalkyl" means a monocyclic or polycyclic saturated or partially unsaturated non-aromatic hydrocarbon. In some embodiments, unless otherwise described, cycloalkyl contains 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. In some embodiments, cycloalkyl is a saturated monocyclic or polycyclic hydrocarbon. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0067] The term "cycloalkylalkyl" means an alkyl group in which at least one hydrogen atom of the alkyl group has been replaced by a cycloalkyl group. Examples of cycloalkylalkyl include cyclopropylmethyl and cyclobutylmethyl. A specific cycloalkylalkyl is cyclopropylmethyl.

[0068] The term "cyano" means the -C≡N group.

[0069] The term "heterocycle" means a monocyclic or bicyclic system of a monovalent saturated or partially unsaturated 4- to 9-membered ring, which contains 1, 2, or 3 heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. Examples of monocyclic saturated heterocycles are azetidinyl, diazepanyl, pyrrolidinyl, tetrahydrofuryl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, and piperazinyl. Examples of polycyclic saturated heterocycles are azaspiroheptyl, diazaspiroheptyl, azaspirooctyl, diazospirooctyl, diazaspirononyl, oxaazaspirooctyl, and oxadiazaspirononyl. A specific example of a heterocycle is piperidinyl. Another specific example of a heterocycle is oxetanyl.

[0070] The term "heteroaryl", alone or in combination, means a monocyclic or bicyclic system of a monovalent aromatic heterocycle of 5 to 12 ring atoms, which contains 1, 2, 3, or 4 heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. Examples of heteroaryl groups include pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrazolyl, pyridinyl, pyridazinyl, pyrimidinyl, triazinyl, azaindolyl, diazaindolyl, isoxazolyl, benzofuryl, isothiazolyl, benzothienyl, indolyl, isoindolyl, isobenzofuryl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, and benzothienyl. A specific example of a heteroaryl group is oxazole.

[0071] The term "heteroarylalkyl" denotes an alkyl group in which one of the hydrogen atoms of the alkyl group is replaced by a heteroaryl group. Examples are oxazolylethyl, tetrazolylethyl, and oxadiazolylethyl. A specific example is oxazolylethyl. Other examples are pyrazolylethyl, tetrazolylpropyl, oxazolylpropyl, and tetrazolylethyl.

[0072] The term "hydroxy" or "hydroxyl" denotes the -OH group.

[0073] The term "hydroxyalkyl" denotes an alkyl group in which at least one hydrogen atom of the alkyl group has been replaced by a hydroxy group. Examples of hydroxyalkyl include hydroxymethyl, hydroxyethyl, hydroxypropyl, 2-hydroxy-1-propyl, 2-hydroxy-2-methyl-1-propyl, 3-hydroxy-1-propyl, and the like. Specific examples of hydroxyalkyl are hydroxyethyl and hydroxypropyl.

[0074] The term "pharmaceutically acceptable salt" refers to those salts that retain the biological effects and properties of the free base or free acid and are not otherwise undesirable biologically. These salts are formed with inorganic acids such as trifluoroacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid (especially hydrochloric acid) and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine. Additionally, these salts can be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, including substituted amines of naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins. The compounds of formula I can also exist in zwitterionic form. Particularly preferred pharmaceutically acceptable salts of the compounds of formula I are salts formed with formic acid and salts formed with hydrochloric acid, yielding hydrochloride, dihydrochloride or trihydrochloride.

[0075] The abbreviation uM means micromole and is equivalent to the symbol μM.

[0076] The abbreviation uL means microliter and is equivalent to the symbol μL.

[0077] The abbreviation ug means microgram and is equivalent to the symbol μg.

[0078] The compounds of formula I can contain several asymmetric centers and can exist as optically pure enantiomers, mixtures of enantiomers (e.g., racemates), optically pure diastereomers, mixtures of diastereomers, diastereomeric racemates or mixtures of diastereomeric racemates.

[0079] According to the Cahn-Ingold-Prelog convention, an asymmetric carbon atom can be of the "R" or "S" configuration.

[0080] Another embodiment of the present invention provides a compound of formula I as described herein and its pharmaceutically acceptable salts or esters, particularly a compound of formula I as described herein and its pharmaceutically acceptable salts, and more particularly a compound of formula I as described herein.

[0081] One embodiment of the present invention provides a compound of formula I as described herein, wherein R 1 and R 5 form, with the atoms to which they are bonded, a 4- to 6-membered heterocycle containing a single O heteroatom.

[0082] One embodiment of the present invention provides a compound of formula I as described herein, wherein R 1 and R 5 form, with the atoms to which they are bonded, a 4- to 5-membered heterocycle containing a single O heteroatom.

[0083] One embodiment of the present invention provides a compound of formula I as described herein, wherein R 1 and R 5 form, with the atoms to which they are bonded, a 5-membered heterocycle containing a single O heteroatom.

[0084] One embodiment of the present invention provides a compound of formula I as described herein, wherein R 2 is H and R 3 is alkyl.

[0085] One embodiment of the present invention provides a compound of formula I as described herein, wherein R 2 is H and R 3 is methyl.

[0086] One embodiment of the present invention provides a compound of formula I as described herein, wherein R 4 is selected from

[0087] i. alkyl,

[0088] ii. hydroxyalkyl,

[0089] iii. alkoxyalkyl,

[0090] iv. cycloalkyl substituted with hydroxy or alkoxy,

[0091] v. cycloalkylalkyl substituted with hydroxy, alkoxy or –COOH,

[0092] vi. a 4- to 6-membered heterocycle containing a single O heteroatom,

[0093] vii. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 2 or 3,

[0094] viii. Heteroarylalkyl, where the heteroaryl in the heteroarylalkyl is oxazole heteroaryl,

[0095] ix. Heteroarylalkyl, where the heteroaryl in the heteroarylalkyl is pyrazole,

[0096] x. -(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3,

[0097] xi. -(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3,

[0098] R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form

[0099] a. A 5-membered heterocycle, where the heterocycle is optionally substituted with OH; or

[0100] b. A 6-membered heterocycle additionally containing 1 O heteroatom;

[0101] where when R 6 is –OH, R 4 can only be alkyl.

[0102] One embodiment of the present invention provides a compound according to formula (I) as described herein, where R 4 is selected from

[0103] i. Alkyl,

[0104] ii. Hydroxyalkyl,

[0105] iii. Alkoxyalkyl,

[0106] iv. Cycloalkylalkyl substituted with hydroxy,

[0107] v. A 4-membered heterocycle containing a single O heteroatom,

[0108] vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 3,

[0109] vii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazole heteroaryl,

[0110] viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole,

[0111] ix. -(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3,

[0112] x. -(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3,

[0113] R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form

[0114] a. A 5-membered heterocycle, wherein the heterocycle is optionally substituted with OH; or

[0115] b. A 6-membered heterocycle additionally containing 1 O heteroatom;

[0116] where when R 6 is -OH, R 4 can only be alkyl.

[0117] An embodiment of the present invention provides a compound of formula I as described herein, wherein R 4 is selected from

[0118] i. Alkyl,

[0119] ii. Hydroxyalkyl,

[0120] iii. Alkoxyalkyl,

[0121] iv. Cycloalkyl substituted with hydroxy or alkoxy,

[0122] v. Cycloalkylalkyl substituted with hydroxy, alkoxy or -COOH,

[0123] vi. A 4- to 6-membered heterocycle containing a single O heteroatom,

[0124] vii. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 2 or 3,

[0125] viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazole heteroaryl.

[0126] where when R 6 is -OH, R4 is limited to an alkyl group.

[0127] One embodiment of the present invention provides a compound according to formula (I) as described herein, wherein R 4 is selected from

[0128] i. an alkyl group,

[0129] ii. a hydroxyalkyl group,

[0130] iii. an alkoxyalkyl group,

[0131] iv. a cycloalkylalkyl group substituted with a hydroxy group,

[0132] v. a 4-membered heterocycle containing a single O heteroatom,

[0133] vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 3,

[0134] vii. a heteroarylalkyl group, wherein the heteroaryl in the heteroarylalkyl group is an oxazole heteroaryl;

[0135] wherein when R 6 is –OH, R 4 is limited to an alkyl group.

[0136] One embodiment of the present invention provides a compound according to formula I as described herein, wherein R 6 is H.

[0137] One embodiment of the present invention provides a compound according to formula I, wherein

[0138] R 1 and R 5 form with the atoms to which they are bonded

[0139] i. a 4- to 6-membered heterocycle containing a single O heteroatom, or

[0140] ii. a 4- to 5-membered cycloalkyl ring;

[0141] R 2 and R 3 are selected from H and alkyl groups, where only one of R 2 or R 3 can be H or an alkyl group;

[0142] R 4 is selected from

[0143] i. an alkoxyalkyl group,

[0144] ii. Cycloalkyl substituted by hydroxy or alkoxy,

[0145] iii. Cycloalkylalkyl substituted by hydroxy or alkoxy, and

[0146] iv. A 4- to 6-membered heterocycle containing a single O heteroatom;

[0147] R 6 is H or –OH;

[0148] and pharmaceutically acceptable salts.

[0149] One embodiment of the present invention provides a compound according to formula I, wherein

[0150] R 1 and R 5 together with the atoms to which they are bonded form

[0151] i. A 4- to 6-membered heterocycle containing a single O heteroatom, or

[0152] ii. A 4- to 5-membered cycloalkyl ring;

[0153] R 2 and R 3 are selected from H and alkyl, wherein only one of R 2 or R 3 can be H or alkyl;

[0154] R 4 is selected from

[0155] i. Cycloalkyl substituted by hydroxy or alkoxy,

[0156] ii. Cycloalkylalkyl substituted by hydroxy or alkoxy, and

[0157] iii. A 4- to 6-membered heterocycle containing a single O heteroatom;

[0158] R 6 is H or –OH;

[0159] and pharmaceutically acceptable salts.

[0160] One embodiment of the present invention provides a compound according to formula I, wherein

[0161] R 1 and R 5 together with the atoms to which they are bonded form

[0162] i. A 4- to 6-membered heterocycle containing a single O heteroatom, or

[0163] ii. A 4- to 5-membered cycloalkyl ring;

[0164] R 2 and R 3Selected from H and alkyl, where R 2 or R 3 Only one of them can be H or alkyl;

[0165] R 4 Selected from

[0166] i. Cycloalkyl substituted with hydroxy or alkoxy, and

[0167] ii. Cycloalkylalkyl substituted with hydroxy or alkoxy;

[0168] R 6 Is H or –OH;

[0169] And pharmaceutically acceptable salts.

[0170] One embodiment of the present invention provides a compound according to formula I, wherein

[0171] R 1 and R 5 Form a 4- to 6-membered heterocycle containing a single O heteroatom with the atoms to which they are bonded;

[0172] R 2 and R 3 Selected from H and alkyl, where R 2 or R 3 Only one of them can be H or alkyl;

[0173] R 4 Selected from

[0174] i. Cycloalkyl substituted with hydroxy, and

[0175] ii. Cycloalkylalkyl substituted with hydroxy;

[0176] R 6 Is H or –OH;

[0177] And pharmaceutically acceptable salts.

[0178] One embodiment of the present invention provides a compound according to formula I, wherein

[0179] R 1 and R 5 Form a 4- to 6-membered heterocycle containing a single O heteroatom with the atoms to which they are bonded;

[0180] R 2 Is H and R 3 Is alkyl;

[0181] R 4 Selected from

[0182] i. Cycloalkyl substituted with hydroxy, and

[0183] ii. Hydroxyl-substituted cycloalkylalkyl;

[0184] R 6 is H or –OH;

[0185] and pharmaceutically acceptable salts.

[0186] One embodiment of the present invention provides a compound according to Formula I, wherein

[0187] R 1 and R 5 form, with the atoms to which they are bonded, a 4- to 6-membered heterocycle containing a single O heteroatom;

[0188] R 2 is H and R 3 is alkyl;

[0189] R 4 is selected from

[0190] i. Hydroxyl-substituted cycloalkyl, and

[0191] ii. Hydroxyl-substituted cycloalkylalkyl;

[0192] R 6 is H;

[0193] and pharmaceutically acceptable salts.

[0194] One embodiment of the present invention provides a compound according to Formula I, wherein

[0195] R 1 and R 5 form, with the atoms to which they are bonded, a 5-membered heterocycle containing a single O heteroatom;

[0196] R 2 is H and R 3 is alkyl;

[0197] R 4 is selected from

[0198] i. alkyl,

[0199] ii. hydroxyalkyl,

[0200] iii. alkoxyalkyl,

[0201] iv. hydroxyl-substituted cycloalkylalkyl,

[0202] v. a 4-membered heterocycle containing a single O heteroatom,

[0203] vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) nC(O)NHCH3, where n is 3,

[0204] vii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazole heteroaryl;

[0205] R 6 is H or –OH,

[0206] wherein when R 6 is –OH, R 4 can only be alkyl;

[0207] and pharmaceutically acceptable salts.

[0208] One embodiment of the present invention provides a compound according to formula I, wherein

[0209] R 1 and R 5 form a 5-membered heterocycle containing a single O heteroatom with the atoms to which they are bonded;

[0210] R 2 is H and R 3 is methyl;

[0211] R 4 is selected from

[0212] i. alkyl,

[0213] ii. hydroxyalkyl,

[0214] iii. alkoxyalkyl,

[0215] iv. cycloalkylalkyl substituted with hydroxy,

[0216] v. a 4-membered heterocycle containing a single O heteroatom,

[0217] vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 3,

[0218] vii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazole heteroaryl,

[0219] viii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole;

[0220] ix. -(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3,

[0221] x. -(CH2) nC(O)NR'R", where n is 3 and both R' and R" are CH3, R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form

[0222] a 5-membered heterocycle, which heterocycle is optionally substituted with OH; or

[0223] a 6-membered heterocycle additionally containing 1 O heteroatom;

[0224] R 6 is H or –OH,

[0225] where when R 6 is –OH, R 4 can only be alkyl;

[0226] and pharmaceutically acceptable salts.

[0227] A specific example of the compound of formula I as described herein is 5-[3-[[(3R)-1-(2-hydroxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol or a pharmaceutically acceptable salt thereof.

[0228] Other specific examples of the compound of formula I as described herein are selected from

[0229] 5-[5-methyl-3-[[racemic-(3R)-1-(2-hydroxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0230] 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0231] 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0232] 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formate;

[0233] 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0234] (3S,5R)-1-Ethyl-5-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-3-ol;

[0235] 5-[5-Methyl-3-[[(3R)-1-(2-methoxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0236] 5-[5-Methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0237] 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyric acid;

[0238] 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl];

[0239] Methyl 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyrate;

[0240] 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N-methyl-butyramide;

[0241] 5-[5-Methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0242] and their pharmaceutically acceptable salts.

[0243] Other specific examples of the compound of formula I as described herein are selected from

[0244] 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N,N-dimethyl-butyramide;

[0245] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-1-pyrrolidin-1-yl-butan-1-one;

[0246] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N-(2-hydroxyethyl)-N-methyl-butyramide;

[0247] 1-(3-Hydroxypyrrolidin-1-yl)-4-[racemic-(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butan-1-one; formic acid;

[0248] 1-(3-Hydroxypyrrolidin-1-yl)-4-[racemic-(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butan-1-one;

[0249] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-1-N-morpholinyl-butan-1-one;

[0250] 5-[5-Methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0251] 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formic acid;

[0252] 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0253] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]

[0254] amino]-1-piperidinyl]butanenitrile;

[0255] 5-[5-Methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0256] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]

[0257] amino]-1-piperidinyl]-N,N-dimethyl-butyramide; formic acid"

[0258] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]

[0259] amino]-1-piperidinyl]-N,N-dimethyl-butyramide; formic acid;

[0260] 5-[5-Methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0261] 5-[5-Methyl-3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0262] and their pharmaceutically acceptable salts.

[0263] More preferred examples of the compound of formula I as described herein are 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyric acid or its pharmaceutically acceptable salts.

[0264] Other more preferred examples of the compound of formula I as described herein are selected from

[0265] 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol;

[0266] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]

[0267] amino]-1-piperidinyl]-N,N-dimethyl-butyramide;

[0268] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]

[0269] amino]-1-piperidinyl]-1-pyrrolidin-1-yl-butan-1-one;

[0270] and its pharmaceutically acceptable salts.

[0271] Another embodiment of the present invention provides a pharmaceutical composition or medicament containing the compound of the present invention and a therapeutically inert carrier, diluent or excipient, and a method for using the compound of the present invention to prepare such compositions and medicaments. In one example, the compound of formula I can be formulated into a galenical administration form by mixing with a physiologically acceptable carrier (i.e., a carrier that is non-toxic to the recipient at the doses and concentrations used) at ambient temperature at an appropriate pH and desired purity. The pH of the formulation depends mainly on the specific use and concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of formula I is formulated in an acetate buffer at pH 5. In another embodiment, the compound of formula I is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.

[0272] The composition is formulated, dosed and administered in a manner consistent with good medical practice. Factors to be considered in this case include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the timing of administration, and other factors known to the practicing physician.

[0273] The compounds of the present invention can be administered by any suitable means, including orally, topically (including buccal and sublingual), rectally, vaginally, transdermally, parenterally, subcutaneously, intraperitoneally, intraluminally, intradermally, intrathecally and epidurally and intranasally, and (if needed for local treatment) intralesionally. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0274] The compounds of the present invention can be administered in any convenient form of administration, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions can contain conventional components in pharmaceutical formulations, for example, diluents, carriers, pH regulators, sweeteners, fillers and other active agents.

[0275] Conventional formulations are prepared by mixing the compounds of the present invention with carriers or excipients. Suitable carriers and excipients are well known to those skilled in the art and are described in detail, for example, in Ansel, Howard C. et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams and Wilkins, 2004; Gennaro, Alfonso R. et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams and Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavoring agents, diluents, and other known additives to provide an aesthetic presentation of the medicament (e.g., the compounds of the present invention or their pharmaceutical compositions) or to facilitate the preparation of the pharmaceutical product (e.g., the drug).

[0276] The compounds of formula I and their pharmaceutically acceptable salts can be processed together with pharmaceutically inert inorganic or organic auxiliaries for the production of tablets, coated tablets, dragees, hard gelatin capsules, injection solutions or topical preparations. For example, lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc. can be used as such auxiliaries for tablets, dragees and hard gelatin capsules.

[0277] Suitable auxiliaries for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semi-solid substances and liquid polyols, etc.

[0278] Suitable auxiliaries for the preparation of solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose, etc.

[0279] Suitable auxiliaries for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils, etc.

[0280] Suitable auxiliaries for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solid or liquid polyols, etc.

[0281] Suitable auxiliaries for topical ophthalmic preparations are, for example, cyclodextrins, mannitol or many other carriers and excipients known in the art.

[0282] In addition, the pharmaceutical preparation may contain preservatives, solubilizers, thickening substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorings, salts for altering the osmotic pressure, buffering agents, masking agents or antioxidants. They may also contain other therapeutically valuable substances.

[0283] The dosage can vary within a wide range and will of course be suitable for the various requirements in each specific case. Generally speaking, in the case of oral administration, a daily dosage of about 0.1 mg to 20 mg per kg body weight, preferably about 0.5 mg to 4 mg per kg body weight (e.g., about 300 mg per person), should be appropriate, which is preferably divided into 1 to 3 separate doses (which may consist of, for example, the same amount). In the case of topical administration, the formulation may contain 0.001% to 15% by weight of the drug, and the required dosage may be 0.1 mg to 25 mg, administered once a day or once a week, or several times a day (2 to 4 times), or several times a week. However, it is obvious that when shown to be applicable, the upper or lower limits given herein may be exceeded.

[0284] One embodiment of the invention is a compound according to formula I as described herein, which is used as a therapeutically active substance.

[0285] One embodiment of the invention is a compound according to formula I as described herein, which is used for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

[0286] One embodiment of the invention is a compound according to formula I as described herein, which is used for treating or preventing a disease, disorder or condition, wherein the disorder or condition responds to NLRP3 inhibition.

[0287] As used herein, the term "NLRP3 inhibition" refers to a complete or partial reduction in the level of NLRP3 activity and includes, for example, inhibiting active NLRP3 and / or inhibiting the activation of NLRP3.

[0288] There is evidence that NLRP3-induced IL-1 and IL-18 play a role in the inflammatory responses associated with or caused by a variety of different diseases (Menu et al., Clinical and Experimental Immunology, 166: 1-15, 2011; Strowig et al., Nature, 481: 278-286, 2012).

[0289] In one embodiment, the disease, disorder or condition is selected from:

[0290] (i) Inflammation;

[0291] (ii) Autoimmune diseases;

[0292] (iii) Cancer;

[0293] (iv) Infection;

[0294] (v) Metabolic diseases;

[0295] (vi) Cardiovascular diseases;

[0296] (vii) Respiratory diseases;

[0297] (viii) Liver diseases;

[0298] (ix) Kidney diseases;

[0299] (x) Eye diseases;

[0300] (xi) Skin diseases;

[0301] (xii) Lymphatic conditions;

[0302] (xiii) Graft-versus-host disease;

[0303] (xiv) Abnormal pain;

[0304] (xv) Conditions related to diabetes; and

[0305] (xvi) Any disease in which it has been determined that the individual carries a germline or somatic non-silent mutation in NLRP3

[0306] In another embodiment, the disease, disorder or condition is selected from:

[0307] (i) Cancer;

[0308] (ii) Infection;

[0309] (iv) Cardiovascular diseases;

[0310] (v) Liver diseases;

[0311] (vi) Respiratory diseases;

[0312] (vi) Eye diseases; and

[0313] (vii) Skin diseases.

[0314] In another embodiment, the disease, disorder or condition is selected from:

[0315] (i) Gout; and

[0316] (ii) Arthritis.

[0317] In yet another exemplary embodiment of the present invention, the disease, disorder or condition is inflammation. Examples of inflammation that can be treated or prevented include inflammatory responses associated with or caused by the following diseases:

[0318] (i) Skin diseases such as contact hypersensitivity, bullous pemphigoid, sunburn, psoriasis, atopic dermatitis, contact dermatitis, allergic contact dermatitis, seborrheic dermatitis, lichen planus, scleroderma, pemphigus, epidermolysis bullosa, urticaria, erythema or alopecia;

[0320] (ii) Arthropathies such as osteoarthritis, systemic juvenile idiopathic arthritis, adult-onset Still's disease, relapsing polychondritis, rheumatoid arthritis, juvenile chronic arthritis, gout or seronegative spondyloarthropathies (such as ankylosing spondylitis, psoriatic arthritis or Reiter's disease);

[0321] disease);

[0322] (iii) Muscle diseases such as polymyositis or myasthenia gravis;

[0323] (iv) Gastrointestinal diseases such as inflammatory bowel disease (including Crohn's disease and ulcerative colitis), colitis, gastric ulcer, celiac disease, proctitis, pancreatitis, eosinophilic gastroenteritis, mastocytosis, antiphospholipid syndrome, or food-related allergies that may affect beyond the gut (such as migraine, rhinitis or eczema);

[0324] (v) Respiratory diseases such as chronic obstructive pulmonary disease (COPD), asthma (including eosinophilic asthma, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma or dust asthma, and especially chronic or refractory asthma such as late-stage asthma and airway hyperreactivity), bronchitis, rhinitis (including acute rhinitis, allergic rhinitis, atrophic rhinitis, chronic rhinitis, caseous rhinitis, hypertrophic rhinitis, pustular rhinitis, dry rhinitis, drug-induced rhinitis, membranous rhinitis, seasonal rhinitis such as hay fever and vasomotor rhinitis), sinusitis, idiopathic pulmonary fibrosis (IPF), sarcoidosis, farmer's lung, silicosis, asbestosis, inflammation caused by volcanic ash, adult respiratory distress syndrome, allergic pneumonia or idiopathic interstitial pneumonia;

[0325] (vi) Vascular diseases such as atherosclerosis, Behçet's disease, vasculitis or Wegener's granulomatosis;

[0326] (vii) Autoimmune diseases such as systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, Hashimoto's thyroiditis, type I diabetes, idiopathic thrombocytopenic purpura or Graves' disease;

[0327] (viii) Other diseases such as sepsis, septic shock, endotoxic shock, toxic shock syndrome, multiple organ failure, acute pancreatitis, acute respiratory distress syndrome, burns, trauma, ischemia-reperfusion injury, transplantation rejection, or any other disease or condition associated with an inflammatory response.

[0328] (ix) Autoimmune diseases such as systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, Hashimoto's thyroiditis, type I diabetes, idiopathic thrombocytopenic purpura or Graves' disease;

[0329] (viii) Ocular diseases such as uveitis, allergic conjunctivitis or vernal conjunctivitis;

[0330] (ix) Infections or infection-related diseases such as acquired immunodeficiency syndrome (AIDS), acute or chronic bacterial infections, acute or chronic parasitic infections, acute or chronic viral infections, acute or chronic fungal infections, meningitis, hepatitis (hepatitis A, B, C or other viral hepatitis), peritonitis, pneumonia, epiglottitis, malaria, dengue hemorrhagic fever, leishmaniasis, streptococcal myositis, Mycobacterium tuberculosis (including Mycobacterium tuberculosis and HIV coinfection), Mycobacterium avium-intracellulare, Pneumocystis carinii pneumonia, orchitis / epididymitis, Legionella, Lyme disease, influenza A, Epstein-Barr virus infection, viral encephalitis / aseptic meningitis or pelvic inflammatory disease;

[0331] (x) Kidney diseases such as mesangial proliferative glomerulonephritis, nephrotic syndrome, nephritis, glomerulonephritis, obesity-related glomerulopathy, acute renal failure, acute kidney injury, uremia, nephrotic syndrome, renal fibrosis (including chronic crystalline nephropathy) or renal hypertension;

[0332] (xi) Lymphatic diseases such as Castleman disease;

[0333] (xii) Diseases of the immune system or diseases involving the immune system such as hyper-IgE syndrome, leprosy, familial hemophagocytic lymphohistiocytosis or graft-versus-host disease; (xiii) Liver diseases such as chronic active hepatitis, non-alcoholic fatty liver disease (NASH), alcoholic hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease (AFLD), alcoholic steatohepatitis (ASH), primary biliary cirrhosis, fulminant hepatitis, liver fibrosis or liver failure;

[0334] (xiv) Cancers, including those listed above;

[0335] (xv) Burns, trauma, injury, bleeding or stroke;

[0336] (xvi) Radiation exposure;

[0337] (xvii) Metabolic diseases such as type 2 diabetes (T2D), atherosclerosis, obesity, gout or pseudogout; and / or

[0338] (xviii) Pain such as inflammatory hyperalgesia, pelvic pain, allodynia, neuropathic pain or bone pain caused by cancer.

[0339] An embodiment of the present invention is a compound according to formula I as described herein, which is useful for treating or preventing a disease, disorder or condition selected from:

[0340] (i) Inflammation;

[0341] (ii) Autoimmune diseases;

[0342] (iii) Cancer;

[0343] (iv) Infections;

[0344] (v) Metabolic diseases;

[0345] (vi) Cardiovascular diseases;

[0346] (vii) Respiratory diseases;

[0347] (viii) Liver diseases;

[0348] (ix) Kidney diseases;

[0349] (x) Eye diseases;

[0350] (xi) Skin diseases;

[0351] (xii) Lymphatic conditions;

[0352] (xiii) Psychological disorders;

[0353] (xiv) Graft-versus-host disease;

[0354] (xv) Neuropathic pain;

[0355] (xvi) Conditions associated with diabetes; and

[0356] (xvii) Any disease in which an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.

[0357] An embodiment of the present invention is the use of a compound according to formula I as described herein in the treatment or prevention of a disease, disorder or condition that responds to NLRP3 inhibition.

[0358] An embodiment of the present invention is the use of a compound according to formula I as described herein in the treatment or prevention of a disease, disorder or condition selected from asthma and COPD.

[0359] An embodiment of the present invention is the use of a compound according to formula I as described herein in the treatment or prevention of a cardiovascular disease, disorder or condition.

[0360] One embodiment of the present invention is the use of a compound of formula I as described herein in the treatment or prevention of a disease, disorder or condition selected from Cryopyrin-associated periodic syndromes.

[0361] One embodiment of the present invention is a compound of formula I as described herein for use in the treatment or prevention of a disease, disorder or condition selected from asthma and COPD.

[0362] One embodiment of the present invention is a compound of formula I as described herein for use in the treatment or prevention of a cardiovascular disease, disorder or condition.

[0363] One embodiment of the present invention is a compound of formula I as described herein for use in the treatment or prevention of a disease, disorder or condition selected from Cryopyrin-associated periodic syndromes.

[0364] One embodiment of the present invention is the use of a compound of formula I as described herein for the preparation of a medicament for the treatment or prevention of a disease, disorder or condition selected from asthma and COPD.

[0365] One embodiment of the present invention is the use of a compound of formula I as described herein for the preparation of a medicament for the treatment or prevention of a cardiovascular disease, disorder or condition.

[0366] One embodiment of the present invention is the use of a compound of formula I as described herein for the preparation of a medicament for the treatment or prevention of a disease, disorder or condition selected from Cryopyrin-associated periodic syndromes.

[0367] One embodiment of the present invention is a method for the treatment or prevention of a disease, disorder or condition selected from asthma and COPD, the method comprising administering an effective amount of a compound of formula I as described herein.

[0368] One embodiment of the present invention is a method for the treatment or prevention of a cardiovascular disease, disorder or condition, the method comprising administering an effective amount of a compound of formula I as described herein.

[0369] One embodiment of the present invention is a method for the treatment or prevention of a disease, disorder or condition selected from Cryopyrin-associated periodic syndromes, the method comprising administering an effective amount of a compound of formula I as described herein.

[0370] One embodiment of the present invention relates to a method of inhibiting NLRP3, the method comprising administering an effective amount of a compound of formula I as described herein.

[0371] Another embodiment of the present invention is a compound of formula I as described herein, which is manufactured according to any one of the said methods.

[0372] One embodiment of the present invention is a pharmaceutical composition comprising a compound according to Formula I as described herein and a therapeutically inert carrier.

[0373] Determination procedure

[0374] NLRP3 and pyroptosis

[0375] It is well known that the activation of NLRP3 leads to pyroptosis, and this feature plays an important role in the manifestation of clinical diseases (Yang-gang Liu et al., Cell Death & Disease, 2017, 8(2), e2579; Alexander Wree et al., Hepatology, 2014, 59(3), 898-910; Alex Baldwin et al., Journal of Medicinal Chemistry, 2016, 59(5), 1691-1710; Ema Ozaki et al., Journal of Inflammation Research, 2015, 8, 15-27; Zhen Xie and Gang Zhao, Neuroimmunology Neuroinflammation, 2014, 1(2), 60-65; Mattia Cocco et al., Journal of Medicinal Chemistry, 2014, 57(24), 10366-10382; T. Satoh et al., Cell Death & Disease, 2013, 4, e644). Therefore, inhibitors of NLRP3 are expected to prevent pyroptosis and the release of pro-inflammatory cytokines (such as IL-1β) from cells.

[0376] THP-1 cells: Culture and preparation

[0377] Grow THP-1 cells (ATCC#TIB-202) in RPMI containing L-glutamine (Gibco#11835) supplemented with 1 mM sodium pyruvate (Sigma#S8636) and penicillin (100 units / ml) / streptomycin (0.1 mg / ml) (Sigma#P4333) in 10% fetal bovine serum (FBS) (Sigma#F0804). Cells are passaged routinely and grown to confluence (about 10 6(cells / ml). On the day of the experiment, THP-1 cells were harvested and resuspended in RPMI medium (without FBS). The cells were then counted and viability was checked by trypan blue (Sigma #T8154) (>90%). Appropriate dilutions were made to obtain a concentration of 625,000 cells / ml. LPS (Sigma #L4524) was added to the diluted cell solution to obtain a final assay concentration (FAC) of 1 μg / ml. 40 μl of the final preparation was aliquoted into each well of a 96-well plate. The plate thus prepared was used for compound screening.

[0378] THP-1 cell pyroptosis assay

[0379] Compound screening was performed by following the stepwise assay method below.

[0380] THP-1 cells (25,000 cells / well) containing 1.0 μg / ml LPS were seeded in 40 μl of RPMI medium (without FBS) in a 96-well, black-walled, clear-bottom cell culture plate coated with poly-D-lysine (VWR #734-0317)

[0381] 5 μl of compound (8-point half-log dilution, 10 μM highest dose) or vehicle (DMSO 0.1% FAC) was added to the appropriate wells

[0382] Incubate at 37 °C, 5% CO2 for 3 hours

[0383] 5 μl of nigericin (Sigma #N7143) (FAC 5 μM) was added to all wells

[0384] Incubate at 37 °C, 5% CO2 for 1 hr

[0385] At the end of the incubation period, spin the plate at 300 x g for 3 minutes and remove the supernatant

[0386] Then 50 μl of resazurin (Sigma #R7017) (FAC 100 μM resazurin, dissolved in RPMI medium without FBS) was added, and the plate was incubated for an additional 1 - 2 hours at 37 °C and 5% CO2. The plate was read in an Envision reader at Ex 560 nm and Em 590 nm

[0387] IC 50 The data fit a non-linear regression equation (log inhibitor vs. response variable slope 4-parameter)

[0388] The results of the pyroptosis assay are summarized in Table 1 below as THP IC 50 .

[0389] Human whole blood IL-1β release assay

[0390] For systemic delivery, the ability to inhibit NLRP3 when the compound is present in the bloodstream is very important. For this reason, the NLRP3 inhibitory activity of various compounds in human whole blood was studied according to the following protocol.

[0391] Human whole blood in lithium heparin tubes was obtained from healthy donors from a panel of volunteer donors.

[0392] 80 μl of whole blood containing 1 μg / ml LPS was placed into a 96-well clear bottom cell culture plate (Corning #3585)

[0393] 10 μl of the compound (8-point half-log dilution, 10 μM highest dose) or vehicle (DMSO 0.1% FAC) was added to the appropriate wells

[0394] Incubated at 37 °C, 5% CO2 for 3 hours

[0395] 10 μl of nigericin (Sigma #N7143) (10 μM FAC) was added to all wells and incubated at 37 °C, 5% CO2 for 1 hr

[0396] At the end of the incubation period, the plate was spun at 300 x g for 5 minutes to pellet the cells and 20 μl of the supernatant was removed and added to a 96-well V-bottom plate for IL-1β analysis (Note: These plates containing the supernatant can be stored at -80 °C for later analysis)

[0397] IL-1β was measured according to the manufacturer's protocol (Perkin Elmer - AlphaLisa IL-1 Kit AL220F-5000)

[0398] IC 50 The data fit a non-linear regression equation (log inhibitor vs. response variable slope 4-parameter)

[0399] The results of the human whole blood assay are summarized in Table 1 below as HWB IC 50 。

[0400] Microsomal stability :

[0401] On a TECAN (Tecan Group Ltd, Switzerland) automated liquid handling system, incubation was carried out at 37 °C in a 96-well plate with 1 μM test compound (0.5 mg / mL) in microsomes plus the cofactor NADPH. After pre-incubating the test compound with microsomes for 10 minutes, the enzymatic reaction was initiated by adding the cofactor. At 1, 3, 6, 9, 15, 25, 35, and 45 minutes, aliquots of the incubation were taken and quenched with 1:3 (v / v) acetonitrile containing an internal standard. The samples were then cooled and centrifuged, and the supernatant was analyzed by LC-MS / MS2.

[0402] Metabolic stability in hepatocytes :

[0403] Assay description:

[0404] Biomaterials. Cryopreserved hepatocytes [from mice, rats, rabbits, monkeys, and humans (male and female; mixed)] were obtained. Throughout the study, the viability after reconstitution of the hepatocytes was at least 80%. Ready-to-use rat / human cultures [long-term hepatocyte co-cultures; pooled (for humans, n = 5 males and n = 5 females)] with stromal mouse fibroblasts (negative control; pooled) and plates, application media, and maintenance media for incubation were used.

[0405] Metabolism was evaluated by suspending hepatocytes. Primary pooled cryopreserved hepatocytes were reconstituted in pre-warmed William's E medium containing 10% FCS, 0.05 mg / mL streptomycin, 50 U / mL penicillin, 0.4 mM L-glutamine, 0.01 mg / mL gentamicin, 0.048 mg / mL hydrocortisone, and 0.004 mg / mL insulin to a final suspension density of 1 × 106 cells / mL. Incubation was carried out fully automatically using a liquid handling system (Tecan) equipped with a CO2 incubator with an orbital shaker. After adding the test compound, e.g., 1 μM, to the wells (1 × 105 cells / well), the 96-well hepatocyte suspension culture plates were incubated at 37 °C in 5% CO2. Samples were quenched by adding acetonitrile (including the internal standard) to the incubation wells at specified time points up to 2 hours.

[0406] Metabolism was evaluated by Incubation of the test article (e.g., 1 μM, 0.1% v / v DMSO) as carried out in the suspension assay was performed in 96-well plates containing co-cultures of adherent hepatocytes and mouse fibroblast control cells or separate control cells (5% CO2 atmosphere and 37 °C). Human The incubation medium was the same as that in the suspension hepatocytes. At defined time points (2, 18, 26, 48, 72, and 96 hours), all wells were quenched with ice-cold acetonitrile containing an internal standard.

[0407] The samples were then centrifuged appropriately and the supernatant was analyzed by LC-MS / MS. The incubation was performed with n = 1 or 2.

[0408] hERG screening assay

[0409] During the development of small molecule drugs, one of the most common adverse side effects leading to drug failure is arrhythmia. Such failures are often associated with the ability of the drug to inhibit the human ether-à-go-go related gene (hERG) cardiac potassium channel. Therefore, no inhibition or low inhibition of the hERG cardiac potassium channel is considered beneficial.

[0410] Cells

[0411] The CHO crelox hERG cell line (ATCC reference number PTA-6812, female Chinese hamster cells) was generated and validated at Roche. Ready-to-use cryopreserved CHO-hERG cells were stored frozen at Evotec (Germany) and used directly for experiments.

[0412] Experimental solutions

[0413] The extracellular solution contained (in mM): NaCl 150; KCl 4; CaCl2 1; MgCl2 1; HEPES 10; pH 7.2 to 7.4 (with NaOH), osmotic pressure 290 mOsm to 330 mOsm. The internal solution contained (in mM): KCl, 10; KF, 100; NaCl, 10; HEPES, 10; EGTA, 20; pH = 7.0 - 7.4 (with KOH), osmotic pressure 260 mOsm to 300 mOsm.

[0414] Electrophysiology

[0415] The effects of the compound on hERG K+ current parameters were evaluated in at least 4 cells at 2 concentrations.

[0416] The hERG assay was performed using an automated patch clamp system 384 (Nanion Technologies GmbH, Germany). The K+ current was measured using patch-voltage-clamp technology in the whole-cell configuration at 35°C to 37°C.

[0417] The cells were held at a resting voltage of -80 mV, by In Figure 1The voltage mode shown (for initiating outward K + current in the pulse mode at 35 °C to 37 °C) stimulates them to activate the hERG channel and conduct the IKhERG current outward, with a stimulation frequency of 0.1 Hz (6 bpm).

[0418] Data analysis

[0419] The amplitudes of IKhERG were recorded at each drug concentration and compared with the vehicle control value (taken as 100%) to define the block fraction. The concentration - response data were fitted with the following relationship:

[0420]

[0421] Using the EworkBook suite (ID Business Solutions Ltd, UK), the concentration - response curve was fitted by non - linear regression analysis. The data fitting was done using a 4 - parameter logistic model (fit = (A+(B / (1+((x / C)^D)))), where A = 0 and B = 100).

[0422] The results of the hERG assay are presented as hERG IC 20 summarized in Table 2 below.

[0423] Transcellular P-gp assay:

[0424] The general assay uses transfected LLC - PK1 cells (porcine kidney epithelial cells) overexpressing human or mouse P - gp, cultured on 96 - well semi - permeable filter plates, where they form a polarized monolayer with tight junctions and act as a barrier between the apical and basolateral compartments.

[0425] P - gp is expressed in the apical membrane of the monolayer. The tightness of the cell monolayer and the functional activity of P - gp are confirmed by adding the cell - impermeable tracer fluorescein and the reference P - gp substrate edoxaban, respectively. J. Pharmacol. Exp Ther., 2021, 376, 322–329.

[0426] PAMPA :

[0427] PAMPA (Parallel Artificial Membrane Permeability Assay) is the first - line permeability screening method for candidate drugs. The PAMPA assay uses an artificial phospholipid membrane to mimic the transcellular absorption conditions. This assay determines the permeability values that can be used for compound optimization and ranking purposes, as well as the input parameters for in - silico simulation models to predict intestinal absorption.

[0428] The donor concentration is measured at t - start (reference) and compared with the donor and acceptor concentrations after a certain time (t - end) to calculate the extent of compound passage through the membrane.

[0429] Bacterial reverse mutation test (AMES):

[0430] Testing of the compound was carried out as outlined in this guideline: Test No. 471: Bacterial Reverse Mutation Assay | OECD Guidelines for the Testing of Chemicals, Section 4: Health Effects | OECD iLibrary (oecd-ilibrary.org)

[0431] Bacterial cultures:

[0432] The bacterial strains used were TA98, TA100, TA1535, TA97a, and TA102. Batches of each strain were stored as frozen stocks. Vials were thawed and used to inoculate cultures in nutrient broth. The cultures were placed in an incubator set at 37 °C and agitated for approximately 10 hours to provide a working culture of at least 108 cells per mL.

[0433] To ensure that the cultures were in the appropriate growth phase and culture density, samples were taken from each culture at the end of the incubation period and the culture density was evaluated by viability plating or OD650 assessment.

[0434] Treatment:

[0435] Three replicates of each concentration of the compound and positive control and six replicates of each vehicle control were included.

[0436] DMSO was used to prepare the formulations to allow for a maximum exposure up to the solubility limit of the test item or 1000 μg / well. This concentration is equivalent to 5000 μg / plate as used in the standard plate incorporation Ames assay.

[0437] In a single experiment, concentrations were usually separated at semi-logarithmic intervals. For soluble compounds, the concentrations were 0, 3.2, 10, 32, 100, 320, 1000 μg / well.

[0438] The positive controls used were:

[0439] Abbreviated names were used for the strains

[0440] 2NF 2-Nitrofluorene TA98 – S-9

[0441] NaN3 Sodium azide TA100 and TA1535 – S-9

[0442] AAC 9-Aminoacridine TA97a – S-9

[0443] MMC Mitomycin C TA102 – S-9

[0444] B[a]P Benzo[a]pyrene TA98 + S-9 +

[0445] AAN aminoanthracene TA100, TA1535, TA97a and TA102 + S-9

[0446] Plating will be achieved by adding to 400 μL of supplemented molten agar at 45 ± 1 °C in the following order:

[0447] · 20 μL of bacterial culture

[0448] · 20 μL of test article solution / carrier control / positive control

[0449] · 100 μL of 10% S-9 mixture or buffer solution

[0450] Subsequently, mix quickly and pour onto the mutation plate (well).

[0451] After standing, invert the plate and incubate in the dark in an incubator set at 37 °C for 2 to 3 days.

[0452] Toxicity:

[0453] Toxicity is detected by the following parameters:

[0454] · Background reduction

[0455] · A significant reduction in mutant revertants compared to the concurrent carrier control

[0456] · Reduction in mutagenic response.

[0457] Scoring:

[0458] The bacterial colonies are scored manually or electronically using an automatic colony counter.

[0459] In vitro mammalian cell micronucleus test:

[0460] Testing of the compound is carried out as outlined in this guideline: Test No. 487: In vitro mammalian cell micronucleus test | OECD Guidelines for the Testing of Chemicals, Section 4: Health Effects | OECD iLibrary (oecd-ilibrary.org)

[0461] Cell culture:

[0462] In a humidified incubator set at 37 °C and 5% (v / v) CO2 in air, the cultures were maintained in tissue culture flasks containing HEPES-buffered RPMI 1640 medium with GlutaMAX-1, which medium included 10% (v / v) heat-inactivated fetal bovine serum, 100 units / mL / 100 μg / mL penicillin / streptomycin. Prior to treatment, the cells were passaged at least once at low to medium density. One day before treatment, the cells were passaged at a density of approximately 7 x 104 cells / mL. Prior to treatment, the cells were maintained at 37 °C, 5% (v / v) CO2 in air, in a humid environment.

[0463] Treatment:

[0464] The cultured human lymphoblastoid TK6 cells were exposed to the compound for 3 hours in the presence of S-9, followed by a 24-hour recovery period. In addition, a 27-hour treatment in the absence of S-9 was also included, as many chemicals have been reported to exert their positive effects only after long-term treatment. This is equivalent to approximately 1.5 to 2.0 times the average generation time of the TK6 cells used in this laboratory (cell cycle is approximately 15 hours). All cultures were sampled 27 hours after the start of treatment.

[0465] Dilutions will be prepared in DMSO, which allows a maximum exposure up to the solubility limit (1 mM or 500 μg / mL, whichever is lower).

[0466] Typically, at least 12 concentrations are separated at intervals of 0.7-fold, starting from the upper limit (for soluble compounds with MW ≥ 500, the concentrations will be 9.887, 14.12, 20.18, 28.82, 41.18, 58.82, 84.04, 120.1, 171.5, 245, 350, and 500 μg / mL). The final concentration of DMSO will be 1% v / v. The positive controls are narcotine in the absence of S-9 and cyclophosphamide in the presence of S-9. In 96-well plates, each treatment will include 2 replicates of the compound at each concentration and multiple concurrent vehicle and positive controls, and will be incubated at 37 °C, 5% (v / v) CO2 for the treatment time. The 3-hour treatment cultures will be washed once and re-incubated with fresh medium for 24 hours.

[0467] Harvest:

[0468] At the determined sampling time, an aliquot of the cell suspension will be taken from the designated culture for determining the cell number by using a Coulter counter. The culture designated for analysis will be centrifuged at approximately 200 g for 5 minutes. The cells will be resuspended in 0.075 M KCl and then fixed in fresh, cold methanol / acetic acid (7:1 v / v). Before preparing the slides, the fixed cells will be stored in the fixative at 2 °C to 8 °C.

[0469] The slides will be air-dried and then stained by immersing them in 12.5 μg / mL acridine orange (pH 6.8) in phosphate-buffered saline (PBS) for approximately 10 minutes, followed by washing with PBS (and stirring) for a few seconds.

[0470] Selection of cytotoxicity readings and concentrations:

[0471] The toxicity will be expressed as the population doubling value (PD) relative to the vehicle control. For each concentration, PD is calculated as follows:

[0472] PD = [log(N / X0)] / log 2

[0473] where N = the average final cell count per culture at each concentration

[0474] X0 = the starting (baseline) count

[0475] The highest concentration for micronucleus analysis should not exceed (approximately) 50% cytotoxicity, the highest concentration tested, or the lowest precipitate concentration observed visually at the end of the treatment incubation period. Slides from the highest selected concentration and at least two lower concentrations will be analyzed such that a range of cytotoxicities from maximal to low or none is covered, where appropriate. Micronuclei will be analyzed in at least 1000 mononucleated cells (2000 per concentration) from each culture.

[0476] Evaluation criteria:

[0477] The compound will be considered to induce clastogenic and / or aneuploid events if any of the following occur: - The frequency of MNMON cells is observed to increase significantly statistically at one or more concentrations.

[0478] - In two replicates, the incidence of cells with micronuclei at such concentrations exceeds the normal range.

[0479] - An increase in the proportion of cells with micronuclei is observed that is concentration-related (positive trend test).

[0480] If all of the above criteria are met, the compound will be considered positive in the assay. If none of the above criteria are met, the compound will be considered negative in the assay. Results that only partially meet the above criteria will be handled on a case-by-case basis, but in the context of a screening study, a positive, negative, or ambiguous conclusion will be drawn. Evidence of concentration-related effects is considered useful but not required when evaluating positive results. Biological relevance will be considered, such as the consistency of responses within and between concentrations and (if applicable) between experiments, or effects that occur only at extremely toxic concentrations.

[0481] Pharmacokinetic characteristics of the test substance in minipigs in vivo:

[0482] After intravenous and oral administration, the pharmacokinetics of the test substance were determined in minipigs. The experimental design consisted of three male minipigs, with each animal receiving a single intravenous bolus dose and a single oral dose of the test item. The intravenous dose was administered at a nominal dose volume of 1 mL / kg. The oral dose was administered by gavage at a nominal dose volume of 5 mL / kg. There was a washout period of at least 7 days between the last sampling time and the next dosing time for the same animal. The content of all formulations was within the required range of 85% to 115% of the nominal content. After dosing, blood samples (1 mL) were withdrawn from the saphenous vein (via cannula) or jugular vein of each animal before dosing, at 5, 15, 30 minutes, 1, 2, 4, 8, 24 hours after IV dosing, and before dosing, at 15, 30 minutes, 1, 2, 4, 6, 8, 24, 48 hours after oral dosing. Hematocrit was determined at all time points. The blood:plasma partition factor was determined at the 2-hour and 4-hour time points, and urine was collected as a single sample within 24 hours after dose administration. Blood samples (labeled 1 mL) were withdrawn from the saphenous vein (via cannula) or jugular vein of each animal into polypropylene tubes containing K2EDTA anticoagulant and centrifuged (1500 g, 10 minutes, 4 °C) to prepare plasma for analysis. The remaining blood cells were discarded. Before analysis using a specific LC-MS method, the plasma vials were capped and stored on wet ice for no more than 60 minutes, then transferred to a storage room at <-50 °C (labeled -80 °C).

[0483] Toxicity assessment of the test substance in minipigs in vivo

[0484] The maximum tolerated dose (MTD) of the test item was determined after once-daily oral (gavage) administration to minipigs. Then the toxicity of daily repeated administration for 14 days was evaluated. In addition, the toxicokinetic characteristics of the test item were characterized. Obtained from Ellegaard in Dalmose, Denmark Sufficient specially bred Miniature pigs (animals: age range of 2 to 3 months and weight range of 4 to 6 kg). At the start of dosing, the animals were 4 to 5 months old and had a weight range of 6 to 9.5 kg. A dose volume of 10 mL / kg was used. Individual dose volumes were calculated from the most recent weight of each animal to the target dose levels of 30, 100, and 300 mg / kg / day or other levels depending on non-MTD results. Blood samples were collected on Day 1 and Day 14 for determination of drug concentration in plasma and derived toxicokinetic parameters. The animals were not fed on the scheduled necropsy day. Each animal was anesthetized by intramuscular injection of Zoletil mixture and then exsanguinated. All tissues were preserved in an appropriate fixative. Further analyses included food consumption, body weight, clinical pathology, and complete histopathological examination of target organs.

[0485] Table 1: NLRP3 inhibitory activity

[0486]

[0487]

[0488] Table 2: hERG inhibition assay

[0489]

[0490] Table 3: Transcellular human P-gp assay:

[0491]

[0492]

[0493] *ER = efflux ratio

[0494] The present invention will now be illustrated by the following examples, which are not limiting.

[0495] In the case where the preparation example is obtained as a mixture of enantiomers or diastereoisomers, the pure enantiomers or diastereoisomers can be obtained by the methods described herein or methods known to those skilled in the art such as chiral chromatography or crystallization.

[0496] Experimental methods

[0497] Abbreviations:

[0498]

[0499]

[0500] Examples

[0501] All examples and intermediates were prepared under a nitrogen atmosphere unless otherwise stated.

[0502] Example 1:

[0503] 5-[3-[[(3R)-1-(2-Hydroxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0504]

[0505] Step A: 4-Benzyloxy-5-bromo-2,3-dihydrobenzofuran

[0506] To a solution of 5-bromochroman-4-ol (CAS# 2279149-27-6, 4.59 g, 20.26 mmol, 1.00 eq) in acetonitrile (40 mL) was added potassium carbonate (5.6 g, 40.51 mmol, 2.00 eq), followed by benzyl bromide (4.89 g, 3.4 mL, 28.57 mmol, 1.41 eq). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was extracted with ethyl acetate and water. The aqueous layer was back-extracted with ethyl acetate. The organic layer was washed with water and brine. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 220 g, gradient 0% to 10% ethyl acetate in heptane) to afford the title compound as a colorless oil (6.17 g, 95% yield). LCMS: m / z 305.1 / 307.0 [M+H] + , ESIpos.

[0507] Step B: 2-(4-Benzyloxy-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane Step C: 5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolane-2-yl)-2,3-dihydrobenzofuran-4-ol

[0508]

[0509] To a solution of 4-benzyloxy-5-bromo-2,3-dihydrobenzofuran (Example 1, Step A) (6.16 g, 19.18 mmol, 1.00 eq) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS# 61676-62-8, 5.47 g, 6.0 mL, 29.41 mmol, 1.53 eq) in tetrahydrofuran (80 mL) at -76 °C was added dropwise a 1.6 M solution of n-butyllithium in hexanes (19 mL, 30.4 mmol, 1.59 eq). The mixture was stirred at -76 °C for 2.5 h. The reaction mixture was warmed to -60 °C, quenched at -60 °C with saturated aqueous NH4Cl, warmed to room temperature, and then extracted with ethyl acetate and saturated aqueous NH4Cl. The aqueous layer was back-extracted with ethyl acetate. The organic layers were washed with brine. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 120 g, gradient 0% to 10% ethyl acetate in heptane) to afford the title compound as a colorless oil (5.78 g, 81% yield). LCMS: m / z 353.1 [M+H] + , ESIpos.

[0510] Step D: tert-Butyl (3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate Step E: 6-Chloro-5-methyl-N-[(3R)-3-piperidinyl]-1,2,4-triazin-3-amine

[0511]

[0512] A solution of 2-(4-benzoyloxydihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Example 1, Step B) (5.77 g, 15.56 mmol, 1.00 eq) in ethyl acetate (70 mL) was evacuated three times and flushed with argon. Palladium on activated carbon (10% Pd based) (577 mg, 0.54 mmol, 0.03 eq) was added. The reaction flask was evacuated, flushed with argon, evacuated, and flushed with hydrogen. At room temperature, the reaction mixture was stirred under a hydrogen atmosphere (balloon) for 3 h. Methanol (10 mL) was added. The reaction flask was evacuated and flushed with argon three times, evacuated and then flushed with hydrogen. At room temperature, the reaction mixture was stirred under a hydrogen atmosphere (balloon) for 1 h. The reaction mixture was filtered and washed thoroughly with ethyl acetate / methanol. The filtrate was concentrated in vacuo to afford the title compound as an off-white solid (4.22 g, 98% yield), which was used without further purification. LCMS: m / z 263.2 [M+H] + , ESIpos.

[0513] Step F: 2-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]ethanol

[0514]

[0515] To a mixture of 3,6-dichloro-5-methyl-1,2,4-triazine (CAS# 132434-82-3, 180 mg, 1.10 mmol, 1.00 eq) and tert-butyl (3R)-3-aminopiperidine-1-carboxylate (CAS# 188111-79-7, 330 mg, 1.65 mmol, 1.50 eq) in 1,4-dioxane (3.6 mL) was added N,N-diisopropylethylamine (148 mg, 0.20 mL, 1.15 mmol, 1.04 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was extracted with dichloromethane and water. The organic layer was washed with brine. The aqueous layer was back-extracted twice with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 12 g, gradient of heptane solution of 0% to 40% ethyl acetate). All fractions containing the product were combined and concentrated in vacuo to afford the title compound as a yellow oil (351 mg, 93% yield). LCMS: m / z 328.3 [M+H] + , ESI pos.

[0516] Step G: 5-[3-[[(3R)-1-(2-Hydroxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0517]

[0518] To a solution of tert-butyl (3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate (Example 1, Step D) (150 mg, 0.43 mmol, 1.00 eq) in dichloromethane (1.4 mL) and methanol (0.70 mL) was added dropwise 4 M HCl in dioxane (1.32 g, 1.1 mL, 4.4 mmol, 10.1 eq). The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo. The residue was dissolved in dichloromethane / methanol (9:1), basified with saturated aqueous NaHCO3 and extracted three times with a mixture of dichloromethane / methanol (9:1). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to afford the title compound as an orange foam (95 mg, 91% yield). LCMS: m / z 228.1 [M+H] + , ESI pos.

[0519] Step A: Methyl 2-benzyloxypropionate

[0520]

[0521] To a solution of 6-chloro-5-methyl-N-[(3R)-piperidin-3-yl]-1,2,4-triazin-3-amine (Example 1, Step E) (95 mg, 0.40 mmol, 1.00 eq) in tetrahydrofuran (1.6 mL) was added 2-iodoethanol (CAS# 624-76-0, 84 mg, 0.038 mL, 0.49 mmol, 1.23 eq), followed by N,N-diisopropylethylamine (133 mg, 0.180 mL, 1.03 mmol, 2.60 eq). The reaction mixture was stirred at 40 °C for 16 h. The reaction mixture was cooled to room temperature, quenched with saturated aqueous NaHCO3, and then extracted with ethyl acetate. The aqueous layer was back-extracted with ethyl acetate. The organic layer was washed with brine. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 12 g, gradient 0% to 10% methanol in dichloromethane) to afford the title compound (61 mg, 54% yield) as a pale yellow oil. LCMS: m / z 272.2 [M+H] + , ESI pos.

[0522] Step B: 2-(Benzyloxy)propanal (CAS: 53346-05-7) Step C: Benzyl N-[(3R)-1-(2-benzyloxypropyl)-3-piperidinyl]carbamate

[0523]

[0524] A mixture of 2-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidin-1-yl]ethanol (Example 1, Step F) (56 mg, 0.20 mmol, 1.00 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (87 mg, 0.32 mmol, 1.61 eq), cesium carbonate (192 mg, 0.59 mmol, 3.01 eq), and XPhos Pd G3 (25 mg, 0.03 mmol, 0.15 eq) in 1,4-dioxane (1.2 mL) and water (0.30 mL) was flushed with argon and stirred at 100 °C for 2.5 h. The reaction mixture was cooled to room temperature and extracted with ethyl acetate and semi-saturated NH4Cl solution. The aqueous layer was back-extracted with ethyl acetate. The organic layer was washed with water and brine. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 12 g, gradient: 0% to 100% (dichloromethane:methanol:NH4OH 9:1:0.05) in dichloromethane) to afford the title compound as a yellow foam (49 mg, 64% yield). LCMS: m / z 372.3 [M+H]+ , ESIpos。

[0525] Example 2:

[0526] 5-[5-Methyl-3-[[(3R)-1-(2-hydroxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0527]

[0528] Step D: (3R)-1-(2-(Benzyloxy)propyl)piperidin-3-amine

[0529] To a solution of methyl 2-hydroxypropionate (CAS: 547-64-8; 5.0 g, 48.03 mmol, 1.0 eq) in THF (90 mL) was added NaH (2882 mg, 60% in mineral oil, 72.05 mmol, 1.5 eq) and the mixture was stirred at 0 °C for ten minutes, then benzyl bromide (CAS: 100-39-0; 5.71 mL, 48.0 mmol, 1.0 eq) was added and the reaction mixture was warmed to room temperature and stirred for 1 h. The mixture was quenched with ice-cold aqueous NH4Cl solution (150 mL) and extracted with EtOAc (100 mL x 2). The organic phase was washed with brine (100 mL×2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate, 9:1 to 4:1) to give the title compound as a colorless oil (6.9 g, yield 74%). 1 1H NMR (400 MHz, DMSO-d6) δ [ppm]: 7.35 - 7.2 (m, 5H), 4.57 (d, 1H), 4.44 (d, 1H), 4.13 (q, 1H), 3.67 (s, 3H), 1.33 (d, 3H).

[0530] Step E: 1-[(3R)-3-Amino-1-piperidinyl]propan-2-ol

[0531] At -78 °C under N2, DIBAl-H (7.72 mL, 7.72 mmol, 1.5 eq) was added dropwise to a solution of methyl 2-benzyloxypropionate (1.0 g, 5.15 mmol, 1.0 eq) in DCM (40 mL) over 10 minutes and the reaction mixture was stirred at -78 °C for 1 h. When the reaction was complete, the mixture was quenched with ice-cold NH4Cl (100 mL) and extracted with DCM (100 mL x 2). The organic phase was washed with brine (100 mL×2), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate, 4:1) to give the title compound as a colorless oil (320.0 mg, yield 38%).1 HNMR (400 MHz, DMSO-d6) δ [ppm]: 9.64 (d, 1H), 7.37 - 7.31 (m, 5H), 4.62 - 4.55 (m, 2H), 4.52 - 4.04 (m, 1H), 1.23 (d, 3H).

[0532] Step F: 1-((R)-3-((6-chloro-5-methyl-1,2,4-triazin-3-yl)amino)piperidin-1-yl)propan-2-ol

[0533] To a mixture of N-[(3R)-3-piperidinyl]carbamic acid benzyl ester (CAS: 478646-32-1; 228 mg, 0.97 mmol, 0.5 eq), acetic acid (117 mg, 1.95 mmol, 1.0 eq) in DCE (4 mL) / methanol (2 mL) was added 2-benzyloxypropanal (CAS: 53346-05-7; 320.0 mg, 1.95 mmol, 1.0 eq) and the mixture was stirred for 10 minutes, then NaBH(OAc)3 (2065 mg, 9.74 mmol, 5.0 eq) was added and the mixture was stirred at 25 °C under N2 for 2 h. When the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by column chromatography (SiO2, EA / MeOH: 4 / 1) to afford the title compound as a colorless oil (210 mg, yield 28%). LCMS: m / s 383.4 [M+H]+, ESI pos.

[0534] Step G: 5-[5-Methyl-3-[[(3R)-1-(2-hydroxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0535] At 25 °C, to a solution of N-[(3R)-1-(2-benzyloxypropyl)-3-piperidinyl]carbamic acid benzyl ester (210.0 mg, 0.55 mmol, 1.0 eq) in methanol (4 mL) was added Pd / C (10 mg, 10% palladium on carbon) and the mixture was stirred under H2 (1100 mmHg) for 2 h. The suspension was filtered through a Celite pad and the pad was washed with MeOH (3 mL × 2), and the combined filtrates were concentrated in vacuo to afford the title compound as a yellow oil (136.0 mg, quantitative). LCMS: 249.2 [M+H]+, ESI, pos.

[0536] Step A: Benzyl N-[(3R)-1-(3-benzyloxypropyl)-3-piperidinyl]carbamate

[0537] At 70 °C, a solution of (3R)-1-(2-(benzyloxy)propyl)piperidin-3-amine (136.0 mg, 0.55 mmol, 1.0 eq), Pd / C (10 mg, 10% palladium on carbon), and Pd(OH)2 / C (10 mg, 10% palladium on carbon) in methanol (3 mL) was stirred at 4500 mmHg for 16 h. The suspension was filtered through a Celite pad, and the pad was washed with MeOH (5 mL × 2). The combined filtrates were concentrated in vacuo to afford the title compound as a yellow oil (86 mg, 84% yield). 1H NMR (400 MHz, DMSO-d6) δ [ppm]: 3.74 - 3.68 (m, 1H), 2.77 - 2.67 (m, 2H), 2.58 - 2.55 (m, 1H), 2.21 - 2.09 (m, 2H), 2.03 - 1.97 (m, 1H), 1.90 - 1.85 (m, 1H), 1.69 - 1.66 (m, 1H), 1.64 - 1.58 (m, 1H), 1.45 - 1.33 (m, 1H), 1.12 - 1.03 (m, 1H), 1.01 (d, 3H).

[0538] Step B: (3R)-1-(3-Benzyloxypropyl)piperidin-3-amine

[0539] To a solution of 1-[(3R)-3-aminopiperidin-1-yl]propan-2-ol (86.0 mg, 0.54 mmol, 1.0 eq) and 3,6-dichloro-5-methyl-1,2,4-triazine (CAS# 132434-82-3, 89.13 mg, 0.54 mmol, 1.0 eq) in 1,4-dioxane (2 mL) was added DIEA (0.16 mL, 0.98 mmol, 1.8 eq). The reaction mixture was stirred at 25 °C for 16 h. When the reaction was complete, the reaction mixture was filtered and concentrated in vacuo. The residue was purified by C18 column chromatography (0.1% NH3·H2O in water / MeCN, MeCN: 54% to 46%) and the eluate was lyophilized to afford the title compound as a yellow oil (27.0 mg, 17% yield). LCMS: m / z 286.1 [M+H] + , ESI pos.

[0540] Step C: 3-[(3R)-3-Amino-1-piperidinyl]propan-1-ol (CAS: 1704948-85-5) Step D: 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]propan-1-ol

[0541] At 25 °C, XPhos Pd G3 (16.01 mg, 0.02 mmol, 0.2 eq) was added to a solution of 1-((R)-3-((6-chloro-5-methyl-1,2,4-triazin-3-yl)amino)piperidin-1-yl)propan-2-ol (27.0 mg, 0.09 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (37.15 mg, 0.14 mmol, 1.5 eq), and CsF (10.98 mg, 0.19 mmol, 2.0 eq) in 1,4-dioxane (1 mL) / water (0.2 mL), and then the mixture was stirred at 95 °C under N2 for 2 h. When the reaction was complete, the reaction mixture was cooled to room temperature, and the combined filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (neutral) to give the title compound as a yellow solid (8.31 mg, yield 21%). 1 1H NMR (400 MHz, CD3OD) δ [ppm]: 7.02 (d, 1H), 6.40 (d, 1H), 4.62 (t, 2H), 4.24 - 4.16 (m, 1H), 3.99 - 3.91 (m, 1H), 3.20 (t, 2H), 3.08 2.95 (m, 1H), 2.84 - 2.76 (m, 1H), 2.74 - 2.50 (m, 1H), 2.46 - 2.35 (m, 3H), 2.32 (s, 3H), 1.92 - 1.84 (m, 2H), 1.81 - 1.67 (m, 1H), 1.64 - 1.57 (m, 1H), 1.16 (dd, 3H).

[0542] Example 3:

[0543] 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0544]

[0545] Step E: 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0546] To a solution of N-[(3R)-3-piperidinyl]carbamic acid benzyl ester (CAS: 478646-32-1; 0.5 g, 2.13 mmol, 1.0 eq) and CH3COOH (12.8 mg, 0.21 mmol, 0.1 eq) in DCE (5 mL) was added 3-benzyloxypropanal (CAS: 19790-60-4; 420.5 mg, 2.56 mmol, 1.2 eq), and the mixture was stirred at 20 °C for 5 min. Then, NaBH(OAc)3 (904.6 mg, 4.27 mmol, 5.0 eq) was added, and the mixture was stirred at 20 °C for 1.5 h. The mixture was quenched with ice water (10 ml), and the aqueous layer was extracted twice with EtOAc (50 mL). The combined organic layers were washed with water (50 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting (ethyl acetate / methanol = 40:1 to 5:1) to give the title compound as a colorless oil (700 mg, 84% yield). LCMS: m / z 383.1 [M+H] + , ESIpos.

[0547] ​

[0548] Under N2, to a solution of N-[(3R)-1-(3-benzyloxypropyl)-3-piperidinyl]carbamic acid benzyl ester (700.0 mg, 1.83 mmol, 1.0 eq) in methanol (2 mL) were added Pd / C (10 mg, 10% palladium on carbon) and Pd(OH)2 / C (10 mg, 10% palladium on carbon). After three replacements with H2, the mixture was stirred at 40 °C for 2 h under 1100 mmHg and H2. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give the title compound as a colorless oil (450 mg, 99% yield). LCMS: m / z 249.0 [M+H] + , ESIpos.

[0549] ​

[0550] To a solution of (3R)-1-(3-benzyloxypropyl)piperidin-3-amine (450.0 mg, 2.01 mmol, 1.0 eq) in methanol (10 mL) was added Pd / C (50 mg, 10% palladium on carbon) under N2. After three replacements with H2, the mixture was stirred at 70 °C for 2 h under 4500 mmHg and H2. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give the title compound as a yellow oil (318 mg, 92% yield).

[0551] ​

[0552] To a solution of 3-[(3R)-3-amino-1-piperidinyl]propan-1-ol (57.9 mg, 0.37 mmol, 1.2 eq) and DIEA (0.08 mL, 0.46 mmol, 1.5 eq) in 1,4-dioxane (1 mL) was added 3,6-dichloro-5-methyl-1,2,4-triazine (50.0 mg, 0.3 mmol, 1.0 eq). The reaction mixture was stirred at 20 °C for 16 h. The above reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by reverse flash chromatography (CombiFlash 0.1% NH3·H2O in water-ACN condition) and then lyophilized to give the title compound as a yellow solid (20.0 mg, yield 22%). LCMS: m / z 286.2 [M+H] + , ESI pos.

[0553] ​ ​

[0554] To a mixture of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (13.8 mg, 0.05 mmol, 1.5 eq) and 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]propan-1-ol (10.0 mg, 0.03 mmol, 1.0 eq) in 1,4-dioxane (0.5 mL) / water (0.1 mL) was added CsF (4.07 mg, 0.07 mmol, 2.0 eq), followed by the addition of XPhos Pd G3 (1.48 mg, 0.0 mmol, 0.05 eq) under N2 at 25 °C. The mixture was stirred at 95 °C for 2 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by reverse phase rapid (CombiFlash 0.1% NH3HCO3 in water-ACN condition) to give the title compound as a yellow solid (9.1 mg, yield 66%). LCMS: m / z 386.3 [M+H] + , ESI pos.

[0555] Example 4:

[0556] 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0557]

[0558] Step A: Methyl 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carboxylate

[0559] To a solution of methyl 1-hydroxycyclopropane-1-carboxylate (5.00 g, 43.06 mmol, 1.0 eq; CAS: 33689-29-1) in DMF (50 mL) was added NaH (2.58 g, 60% w / w in mineral oil, 64.59 mmol, 1.5 eq), and the mixture was stirred at 0 °C for 20 min. 2-(Trimethylsilyl)ethoxymethyl chloride (8.38 mL, 47.36 mmol, 1.1 eq) was added dropwise, and the mixture was stirred at 25 °C for 1 h. The above reaction mixture was diluted with NH4Cl (500 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic phases were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 10 / 1) to give the title compound as a yellow oil (2.10 g, yield 20%). 1 1H NMR (400 MHz, CD3OD) δ [ppm]: 4.79 (s, 2H), 3.71 (s, 3H), 3.67 (t, 2H), 1.30 - 1.25 (m, 4H), 0.90 (t, 2H), 0.03 (s, 9H).

[0560] Step B: (1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropyl)methanol

[0561] To a solution of methyl 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carboxylate (1.10 g, 4.46 mmol, 1.0 eq) in DCM (20 mL) was added dropwise DIBAl-H (13.39 mL, 13.39 mmol, 3.0 eq) at -78 °C under N2, and the mixture was stirred at -78 °C for 1 h. The above reaction mixture was diluted with saturated NH4Cl (100 mL) and DCM (20 mL) at 0 °C, filtered, and the filtrate was extracted with DCM (20 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 10 / 1) to give the title compound as a yellow oil (0.55 g, 2.5 mmol, yield 56%). 1 1H NMR (400 MHz, DMSO-d6) δ [ppm]: 4.68 (s, 2H), 4.60 (t, 1H), 3.57 - 3.49 (m, 4H), 0.85 (t, 1H), 0.75 - 0.68 (m, 2H), 0.58 - 0.52 (m, 2H), 0 (s, 9H).

[0562] Step C: 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carbaldehyde

[0563] At -70 °C, oxalyl chloride (0.28 mL, 3.30 mmol, 2.0 eq) was added to a solution of DMSO (0.23 mL, 3.3 mmol, 2.0 eq) in DCM (7 mL), and the mixture was stirred at -70 °C for 10 min. Then a solution of (1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropyl)methanol (0.360 g, 1.65 mmol, 1.0 eq) in DCM (2 mL) was added dropwise to the above solution, and the mixture was stirred at -70 °C for 0.5 h. TEA (1.8 mL, 12.93 mmol, 7.84 eq) was added and stirring was continued at 0 °C for 30 min. The above reaction mixture was warmed to room temperature, diluted with water (50 mL), and extracted with DCM (30 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 10 / 1) to obtain the title compound as a yellow oil (0.18 g, yield 50%). 1 H NMR (400 MHz, DMSO-d6) δ [ppm]: 9.26 (s, 1H), 4.71 (s, 2H), 3.66 (t, 2H), 1.34 - 1.32 (m, 2H), 1.30 - 1.28 (m, 2H), 0.86 (t, 2H), 0.01 (s, 9H).

[0564] Step E: (R)-tert-Butyl 3-((6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3- yl)amino)piperidine-1-carboxylate

[0565] To (3R)-tert-Butyl 3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate (700 mg, 2.14 mmol, 1.0 eq; prepared as described in Step D of Example 1), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (839 mg, 3.2 mmol, 1.5 eq) and CsF (1.30 g, 8.54 mmol, 4.0 eq) in a solution of 1,4-dioxane (20 mL) and water (4 mL), XPhos Pd G3 (0.362 g, 0.43 mmol, 0.2 eq) was added and stirring was continued at 95 °C under N2 for 3 h. The above reaction mixture was cooled to room temperature, diluted with ethyl acetate (100 mL), filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 1 / 1) to obtain the title compound as a yellow solid (0.750 g, yield 82%). 11H NMR (400 MHz, DMSO-d6) δ [ppm]: 9.38 (s, 1H), 7.53 (s, 1H), 6.95 (d, 1H), 6.38 (d, 1H), 4.57 (t, 2H), 3.88 - 3.82 (m, 2H), 3.65 - 3.30 (m, 1H), 3.15 (t, 2H), 3.10 - 2.74 (m, 1H), 2.20 (s, 3H), 1.99 - 1.93 (m, 1H), 1.81 - 1.71 (m, 1H), 1.63 - 1.50 (m, 1H), 1.47 - 1.42 (m, 1H), 1.34 (s, 9H).

[0566] Step F: (R)-5-(5-Methyl-3-(piperidin-3-ylamino)-1,2,4-triazin-6-yl)-2,3-dihydrobenz ofuran-4-ol

[0567] At 25 °C, a solution of tert-butyl (R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidine-1-carboxylate (0.450 g, 1.05 mmol, 1.0 eq) in DCM (3 mL) / TFA (3.0 mL) was stirred for 1 h. The solvent was removed by a stream of nitrogen to give the crude product, and then the pH was adjusted to about 7 with an NH3·H2O solution. The crude product was purified by reverse flash chromatography (0.1% NH3·H2O, water-MeCN) to give the title compound as a yellow solid (0.346 g, yield 77%). LCMS: m / z 328.2 [M+H] + , ESIpos.

[0568] Step G: (R)-5-(5-Methyl-3-((1-((1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropyl) methyl)piperidin-3-yl)amino)-1,2,4-triazin-6-yl)-2,3-dihydrobenzofuran-4-ol

[0569] At 25 °C, a mixture of 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carbaldehyde (0.171 g, 0.79 mmol, 1.3 eq), AcOH (3.7 mg, 0.06 mmol, 0.1 eq), (R)-5-(5-methyl-3-(piperidin-3-ylamino)-1,2,4-triazin-6-yl)-2,3-dihydrobenzofuran-4-ol (200 mg, 0.61 mmol, 1.0 eq) in DCE (2 mL) was stirred for 10 min, then NaBH3CN (7.67 mg, 1.22 mmol, 2.0 eq) was added and the mixture was stirred at 40 °C for 1 h. The solvent was removed by a stream of nitrogen. The crude product was purified by reverse flash chromatography (0.1% NH3·H2O, water-ACN) to give the title compound as a yellow solid (120 mg, 0.23 mmol, yield 37%). LCMS: m / z 528.4 [M+H] + , ESIpos.

[0570] Step H: 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidinyl]amino]-5-methyl-1,2,4- triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0571] A solution of (R)-5-(5-methyl-3-((1-((1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropyl)methyl)piperidin-3-yl)amino)-1,2,4-triazin-6-yl)-2,3-dihydrobenzofuran-4-ol (40 mg, 0.08 mmol, 1.0 eq) in DCM (1 mL) / TFA (1.0 mL) was stirred at 25 °C for 1 h. The solvent was removed by a stream of nitrogen. Then the pH was adjusted to 7 with NH3·H2O, and the crude product was purified by reverse flash chromatography (0.1% NH3·H2O in water-ACN) to afford the title compound as a yellow solid (9.89 mg, 0.02 mmol, 31% yield). LCMS: m / z 398.3 [M+H] + , ESI pos.

[0572] Example 5:

[0573] 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formate

[0574]

[0575] Step A: Benzyl N-[(3R)-1-(2-methoxyethyl)-3-piperidinyl]carbamate

[0576] To a solution of benzyl N-[(3R)-3-piperidinyl]carbamate (CAS# 478646-32-1, 0.5 g, 2.13 mmol, 1.0 eq), N-ethyl-N-isopropylpropan-2-amine (0.75 mL, 4.27 mmol, 2.0 eq) in acetonitrile (5 mL) was added 2-bromoethyl methyl ether (CAS: 6482-24-2; 0.3 mL, 3.2 mmol, 1.5 eq), and the reaction mixture was stirred at 50 °C for 6 h. When the reaction was complete, the reaction mixture was cooled to room temperature. Ethyl acetate (5 mL) and water (5 mL) were added, and the layers were separated. The aqueous phase was extracted with ethyl acetate (5 mL x 3). The combined extracts were washed with brine (5 mL x 3), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 1 to 1 / 2) to afford the title compound as an orange solid (520.0 mg, 83% yield). LCMS: m / z 293.1 [M+H] + , ESI pos.

[0577] Step B: (3R)-1-(2-Methoxyethyl)piperidin-3-amine

[0578] Under N2, Pd / C (50.0 mg, 10% palladium on carbon) and Pd(OH)2 / C (52.22 mg, 10% palladium on carbon) were added to a solution of benzyl N-[(3R)-1-(2-methoxyethyl)-3-piperidinyl]carbamate (470.0 mg, 1.61 mmol, 1.0 eq) in methanol (5 mL). After replacing with H2 three times, the mixture was then stirred at 20 °C under 1100 mmHg and H2 for 2 hours. When the reaction was complete, the reaction mixture was filtered and the mother liquor was concentrated in vacuo to give the title compound as a yellow oil (240.0 mg, yield 94%). 1 1H-NMR (400 MHz, DMSO-d6) δ [ppm]: 3.39 (t, 2H), 3.22 (s, 3H), 2.77 - 2.71 (m, 1H), 2.67 - 2.61 (m, 1H), 2.60 - 2.54 (m, 1H), 2.44 - 2.39 (m, 2H), 1.91 - 1.84 (m, 1H), 1.72 - 1.67 (m, 1H), 1.67 - 1.62 (m, 1H), 1.59 - 1.52 (m, 1H), 1.40 (s, 1H), 0.95 - 0.85 (m, 1H).

[0579] Step C: 6-Chloro-N-[(3R)-1-(2-methoxyethyl)-3-piperidinyl]-5-methyl-1,2,4-triazin-3-amine

[0580] To a solution of (3R)-1-(2-methoxyethyl)piperidin-3-amine (231.6 mg, 1.46 mmol, 1.2 eq), N-ethyl-N-isopropylpropan-2-amine (0.38 mL, 2.2 mmol, 1.8 eq) in 1,4-dioxane (3 mL) was added 3,6-dichloro-5-methyl-1,2,4-triazine (CAS# 132434-82-3, 200.0 mg, 1.22 mmol, 1.0 eq), and the reaction mixture was stirred at 20 °C for 16 h. When the reaction was complete, the mixture was filtered and concentrated in vacuo. The residue was purified by C18 column chromatography (0.1% TFA in water / acetonitrile, acetonitrile: 30% - 40%). The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 10 / 1), and then by preparative TLC (dichloromethane / methanol: 5 / 1) to give the title compound as a colorless oil (30.0 mg, yield 9%). LCMS: m / z 286.2 [M+H]+, ESI pos.

[0581] Step D: 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin- 6-yl]-2,3-dihydrobenzofuran-4-ol; formate

[0582] At 20 °C under N2, to a solution of 6-chloro-N-[(3R)-1-(2-methoxyethyl)-3-piperidinyl]-5-methyl-1,2,4-triazin-3-amine (30.0 mg, 0.1 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (41.3 mg, 0.16 mmol, 1.5 eq), and CsF (63.8 mg, 0.42 mmol, 4.0 eq) in 1,4-dioxane (1 mL) and water (0.2 mL) was added XPhos Pd G3 (17.8 mg, 0.02 mmol, 0.2 eq). The reaction mixture was stirred at 80 °C for 2 h. When the reaction was complete, the mixture was cooled to room temperature, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex luna C 18 150*25 mm*10 um; conditions: water (formic acid)-acetonitrile; start B: 7; end B: 37; gradient time (min): 10; 100% B hold time (min): 2; flow rate (mL / min) 25) to afford the title compound as a brown gummy solid (18.2 mg, 40% yield). LCMS: m / z 386.1 [M+H] + , ESIpos.

[0583] Example 6:

[0584] (3S,5R)-1-Ethyl-5-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-

[0585] yl]amino]piperidin-3-ol

[0586]

[0587] Step A: (3R,5S)-tert-Butyl 3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-5-hydroxypiperidine-1- carboxylate

[0588] To a solution of 3,6-dichloro-5-methyl-1,2,4-triazine (CAS#132434-82-3, 243 mg, 1.48 mmol, 1.0 eq) in 1,4-dioxane (5 mL) was added (3R,5S)-3-amino-5-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (CAS#1932513-59-1, 396 mg, 1.78 mmol, 1.2 eq) and N,N-diisopropylethylamine (345 mg, 0.47 mL, 2.67 mmol, 1.8 eq), thereby producing a dark brown solution. The reaction mixture was stirred at 23 °C for 60 h. Then, the reaction mixture was quenched with semi-saturated aqueous NH4Cl solution (50 mL) and extracted with ethyl acetate (2 x 50 mL). The organic layer was washed with water (80 mL) and brine (80 mL). The combined organic extracts were dried over sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography (silica gel, 25 g, 0% to 80% ethyl acetate in heptane) to afford the title compound as a pale yellow oil (345 mg, 68% yield).

[0589] LCMS: m / z 344.2 ([{35Cl}M+H] + ), 346.2 ([{37Cl}M+H] + ), ESI pos.

[0590] Step B: (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidin-3-ol; hydrochloride

[0591] To a solution of (3R,5S)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-5-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (Example 6, Step A) (345 mg, 1.0 mmol, 1.0 eq) in dichloromethane (10 mL) and methanol (5 mL) was added dropwise 4 M HCl in 1,4-dioxane (3.01 g, 2.51 mL, 10.03 mmol, 10.0 eq). The reaction mixture was stirred at 23 °C for 16 h. The reaction mixture was concentrated in vacuo to afford the title compound as a light brown foam (312 mg, 100% yield), which was used without further purification. LCMS: m / z 244.1 ([{35Cl}M+H]+), 246.1 ([{37Cl}M+H]+), ESIpos.

[0592] Step C: (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-ethylpiperidin-3-ol

[0593] To a suspension of (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidin-3-ol; hydrochloride (Example 6, Step B) (312 mg, 1.0 mmol, 1.0 eq)) in dichloromethane (15 mL) was added acetaldehyde (CAS# 75-07-0); 110 mg, 0.14 mL, 2.51 mmol, 2.5 eq)), followed by addition of sodium acetate (CAS# 127-09-3, 206 mg, 2.51 mmol, 2.5 eq) and sodium triacetoxyborohydride (CAS# 56553-60-7; 382 mg, 1.8 mmol, 1.8 eq). The reaction mixture was stirred at 0 °C for 5 minutes and at 23 °C for 3 hours. The reaction mixture was carefully basified with saturated NaHCO3 solution (25 mL) and then extracted with dichloromethane (3 x 60 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was adsorbed onto ISOLUTE HM-N and the column was purified by flash chromatography (silica gel, 25 g, gradient: 0% to 100% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to give the title compound as a light brown foam (137 mg, 50% yield). LCMS: m / z 272.1 ([{35Cl}M+H] + ), 274.1 ([{37Cl}M+H] + ), ESIpos.

[0594] Step D: (3S,5R)-1-Ethyl-5-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2, 4-triazin-3-yl]amino]piperidin-3-ol

[0595] A mixture of (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-ethyl-piperidin-3-ol (Example 6, Step C) (84 mg, 0.31 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (154 mg, 0.56 mmol, 1.8 eq) and potassium carbonate (205 mg, 1.48 mmol, 4.8 eq) was dissolved in 1,4-dioxane (2 mL) and water (1 mL). The sealable tube was flushed with argon and 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II) dichloromethane complex (CAS# 95464-05-4, 38 mg, 0.046 mmol, 0.15 eq) was added thereto. After flushing again with argon, the sealed tube was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature and quenched with water (50 mL) and saturated NH4Cl solution (50 mL), then extracted with dichloromethane (3 x 50 mL). The organic layer was washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was absorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 12 g, gradient: 0% to 100% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)), followed by purification by preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 x 30 mm; MeCN / water (+0.1% TEA)) to afford the title compound as a yellow amorphous freeze-dried solid (43 mg, yield 37%). LCMS: m / z 372.2 [M+H] + , ESIpos.

[0596] Example 7:

[0597] 5-[5-Methyl-3-[[(3R)-1-(2-methoxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0598]

[0599] Step A: 2-Methoxypropyl ethanesulfonate

[0600] At 0 °C, ethanesulfonyl chloride (1.73 mL, 18.3 mmol) was added to a solution of 2-methoxypropanol (1.10 g, 12.2 mmol, 1.0 eq) and DIEA (4.72 g, 36.6 mmol, 3.0 eq) in DCM (10 mL), and then the mixture was stirred at 20 °C for 1 h. When the reaction was complete, the above reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 1 / 1) to obtain the title compound as a black solid (2.3 g, yield 98%). 1 1H NMR (400 MHz, CD3OD) δ [ppm]: 4.23 - 4.19 (m, 1H), 4.15 - 4.11 (m, 1H), 3.67 - 3.59 (m, 1H), 3.41 (s, 3H), 3.20 (q, 2H), 1.45 (t, 3H), 1.21 (d, 3H).

[0601] Step B: Benzyl N-[(3R)-1-(2-methoxypropyl)-3-piperidinyl]carbamate

[0602] At 0 °C, 2-methoxypropyl ethanesulfonate (532.2 mg, 2.77 mmol, 1.3 eq) was added to a solution of N-[(3R)-3-piperidinyl]carbamic acid benzyl ester (CAS# 478646-32-1, 500.0 mg, 2.13 mmol, 1.0 eq) and TEA (0.5 g, 4.91 mmol, 2.3 eq) in ACN (5 mL), and then the mixture was stirred at 80 °C for 16 h. The above reaction mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate: 1 / 0 to 1 / 1) to obtain the title compound as a brown oil (190.0 mg, yield 29%). 1 1H NMR (400 MHz, CD3OD) δ [ppm]: 7.34 - 7.28 (m, 5H), 5.06 (s, 2H), 3.67 - 3.61 (m, 1H), 3.55 - 3.51 (m, 1H), 3.34 - 3.31 (m, 3H), 2.90 - 2.82 (m, 1H), 2.70 - 2.59 (m, 1H), 2.48 - 2.43 (m, 1H), 2.31 - 2.15 (m, 1.5H), 2.15 - 2.05 (m, 1.5H), 1.82 - 1.68 (m, 2H), 1.63 - 1.57 (m, 1H), 1.33 - 1.25 (m, 1H), 1.11 (d, 3H).

[0603] Step C: (3R)-1-(2-Methoxypropyl)piperidin-3-amine

[0604] At 25 °C, a solution of benzyl N-[(3R)-1-(2-methoxypropyl)piperidin-3-yl]carbamate (190.0 mg, 0.62 mmol, 1.0 eq), Pd(OH)2 (10 mg, 10% Pd on carbon), and Pd / C (10 mg, 10% Pd on carbon) in methanol (6 mL) was stirred under H2 at 1100 mmHg for 2 h. When the reaction was complete, the suspension was filtered through a Celite pad, and the pad was washed with MeOH (5 mL × 3). The combined filtrates were concentrated in vacuo to afford the desired product as a yellow oil, which was used directly without further purification to give the title compound as a yellow oil (75.0 mg, 70% yield). 1 1H NMR (400 MHz, DMSO-d6) δ [ppm]: 3.23 (s, 3H), 2.72 - 2.67 (m, 1H), 2.62 - 2.55 (m, 2H), 2.36 - 2.29 (m, 2H), 2.16 - 2.10 (m, 1H), 1.93 - 1.87 (m, 1H), 1.71 - 1.63 (m, 2H), 1.61 - 1.56 (m, 1H), 1.43 - 1.34 (m, 2H), 1.04 (d, 3H).

[0605] Step D: 6-Chloro-N-[(3R)-1-(2-methoxypropyl)-3-piperidinyl]-5-methyl-1,2,4-triazin-3-amine

[0606] To a solution of (3R)-1-(2-methoxypropyl)piperidin-3-amine (75.0 mg, 0.44 mmol, 1.0 eq) and DIEA (0.14 mL, 0.78 mmol, 1.8 eq) in 1,4-dioxane (1 mL) was added 3,6-dichloro-5-methyl-1,2,4-triazine (71.4 mg, 0.44 mmol, 1.0 eq), and the mixture was stirred at 25 °C for 16 h. When the reaction was complete, the reaction mixture was quenched with H2O (10 mL), extracted with EtOAc (20 mL × 2), and the organic phase was washed with brine (10 mL × 2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate, 1:1) to afford the title compound as a dark green oil (36.0 mg, 28% yield). LCMS: m / z 300.2 [M+H] + , ESIpos.

[0607] Step E: 5-[5-Methyl-3-[[(3R)-1-(2-methoxypropyl)-3-piperidinyl]amino]-1,2,4-triazin- 6-yl]-2,3-dihydrobenzofuran-4-ol

[0608] At 25 °C under N2, to a solution of 6-chloro-N-[(3R)-1-(2-methoxypropyl)-3-piperidinyl]-5-methyl-1,2,4-triazin-3-amine (18.0 mg, 0.06 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (23.61 mg, 0.09 mmol, 1.5 eq; Example 1, Step C), CsF (6.98 mg, 0.12 mmol, 2.0 eq) in 1,4-dioxane (1 mL) / water (0.2 mL) was added XPhos Pd G3 (10.17 mg, 0.01 mmol, 0.2 eq), and then the mixture was stirred at 95 °C for 2 h. When the reaction was complete, the reaction mixture was cooled to room temperature and the combined filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (instrument: ACS WH-GX-M, column: Waters Xbridge 150*25 mm*5 um, mobile phase: A represents H2O (0.1% NH4HCO3) and B represents acetonitrile, gradient: linear in 10 minutes, B 28% to 58%, flow rate: 30 mL / min, column temperature: R.T. wavelength: 220 nm / 254 nm) to lyophilization to give the title compound as a yellow oil (9.58 mg, yield 36%). LCMS: m / z 400.2 [M+H] + , ESI pos.

[0609] Example 8:

[0610] 5-[5-Methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0611]

[0612] Step A: Benzyl (R)-(1-(oxetan-3-yl)piperidin-3-yl)carbamate

[0613] To a solution of (R)-benzyl piperidin-3-ylcarbamate (CAS# 478646-32-1; 1.0 g, 4.27 mmol, 1.0 eq) in DCE (10 mL) was added oxetan-3-one (CAS# 6704-31-0; 0.370 g, 5.12 mmol, 1.2 eq). The mixture was stirred at 20 °C for 1 hour, then NaBH(OAc)3 (1.18 g, 5.55 mmol, 1.3 eq) was added and the mixture was stirred at 20 °C for 1 hour. The above reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (hexane / EtOAc, 1:0 to 0:1, then EtOAc / MeOH, 1:0 to 5:1) to give the title compound as a white solid (1.20 g, yield 97%). 1 H NMR (400 MHz, CDCl3) δ [ppm]: 7.40 - 7.35 (m, 5H), 5.48 - 5.26 (m, 1H), 5.11 (s, 2H), 4.67 - 4.53 (m, 4H), 3.96 - 3.81 (m, 1H), 3.53 - 3.39 (m, 1H), 2.47 - 2.19 (m, 3H), 1.74 - 1.58 (m, 4H).

[0614] Step B: (R)-1-(oxetan-3-yl)piperidin-3-amine

[0615] Under N2, to a solution of (R)-(1-(oxetan-3-yl)piperidin-3-yl)carbamic acid benzyl ester (200 mg, 0.69 mmol, 1.0 eq) in methanol (4 mL) was added Pd / C (10.0 mg, 10% palladium on carbon). After three replacements with H2, the mixture was stirred at 20 °C and 1100 mmHg for 1 hour. Then the mixture was filtered and the filtrate was concentrated under reduced pressure to give the title compound (100 mg, 93%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ [ppm]: 4.67 - 4.56 (m, 4H), 3.49 - 3.41 (m, 1H), 2.98 - 2.87 (m, 1H), 2.66 - 2.54 (m, 1H), 2.50 - 2.41 (m, 1H), 2.03 (s, 2H), 1.95 - 1.81 (m, 2H), 1.79 - 1.68 (m, 2H), 1.65 - 1.52 (m, 1H), 1.22 - 1.08 (m, 1H).

[0616] Step C: (R)-6-Chloro-5-methyl-N-(1-(oxetan-3-yl)piperidin-3-yl)-1,2,4-triazin-3- amine

[0617] To a solution of DIPEA (0.157 g, 1.22 mmol, 2.0 eq), (R)-1-(oxetan-3-yl)piperidin-3-amine (0.100 g, 0.64 mmol, 1.05 eq) in 1,4-dioxane (1 mL) was added 3,6-dichloro-5-methyl-1,2,4-triazine (CAS# 132434-82-3, 0.100 g, 0.61 mmol, 1.0 eq), and then the mixture was stirred at 20 °C for 12 h. The above reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (hexane / EtOAc, 1:0 to 0:1) to give the title compound as a yellow solid (0.03 g, 0.11 mmol, yield 17%). 1 1H NMR (400 MHz, CD3OD) δ [ppm]: 4.72 - 4.57 (m, 4H), 4.14 - 3.99 (m, 1H), 3.61 - 3.49 (m, 1H), 2.95 - 2.81 (m, 1H), 2.68 - 2.55 (m, 1H), 2.45 (s, 3H), 2.11 - 2.03 (m, 1H), 2.02 - 1.92 (m, 2H), 1.87 - 1.77 (m, 1H), 1.72 - 1.61 (m, 1H), 1.55 - 1.41 (m, 1H).

[0618] Step D: 5-[5-Methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidinyl]amino]-1,2,4-tri azin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0619] At 25 °C under N2, to a solution of (R)-6-chloro-5-methyl-N-(1-(oxetan-3-yl)piperidin-3-yl)-1,2,4-triazin-3-amine (0.015 g, 0.05 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (0.021 g, 0.08 mmol, 1.5 eq; Example 1, Step C), KF (0.015 g, 0.26 mmol, 5.0 eq) in 1,4-dioxane (1 mL) and water (0.2 mL) was added XPhos Pd G3 (9 mg, 0.01 mmol, 0.2 eq), and then the mixture was stirred at 90 °C for 4 h. The above reaction mixture was cooled to room temperature, diluted with MeCN (3 mL), filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Waters xbridge 150*25 mm 10um; conditions: water (NH4HCO3)-ACN; starting B 21, ending B 51; gradient time (min): 10; 100% B hold time (min): 2; flow rate (mL / min) 25) to give the title compound as a yellow solid (4.35 mg, yield 20%). LCMS: m / z 384.1 [M+H] + , ESIpos.

[0620] Example 9:

[0621] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl] amino]-1-piperidinyl]butanoic acid

[0622]

[0623] Step A: tert-Butyl (3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate

[0624] To a solution of 3,6-dichloro-5-methyl-1,2,4-triazine (CAS#132434-82-3, 520 mg, 3.17 mmol, 1.0 eq) in 1,4-dioxane (10 mL) was added (3R)-tert-butyl 3-aminopiperidine-1-carboxylate (CAS#188111-79-7, 762 mg, 3.81 mmol, 1.2 eq) and N,N-diisopropylethylamine (472 mg, 0.97 mL, 3.66 mmol, 1.80 eq). The reaction mixture was stirred at 23 °C for 48 h. The reaction mixture was quenched with semi-saturated NaHCO3 solution (100 mL) and extracted with ethyl acetate (2 x 100 mL). The organic layer was washed with water (100 mL) and brine (100 mL). The combined organic extracts were dried over sodium sulfate, filtered, and evaporated. The residue was purified by flash chromatography (silica gel, 12 g, gradient: 0% to 50% ethyl acetate in heptane) to afford the title compound as a pale yellow gum (815 mg, 74% yield). LCMS: m / z 326.0 ([{35Cl}M-H] - ), 328.0 ([{37Cl}M-H] - ), ESI neg.

[0625] Step B: tert-Butyl (3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3- yl]amino]piperidine-1-carboxylate

[0626] In a sealable tube, a mixture of (3R)-tert-butyl 3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate (Example 9, Step A) (165 mg, 0.48 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (Example 1, Step C) (175 mg, 0.67 mmol, 1.40 eq) and cesium carbonate (467 mg, 1.43 mmol, 3.0 eq) in 1,4-dioxane (4 mL) and water (1 mL) was left under argon and finally XPhos Pd G3 gt (61 mg, 0.72 mmol, 0.15 eq) was added. The reaction mixture was stirred in the sealed tube at 95 °C for 4 h. The reaction mixture was cooled to room temperature, then quenched with water (20 mL) and saturated NH4Cl solution (20 mL), and extracted with ethyl acetate (2 x 40 mL). The organic layer was washed with brine (40 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, 12 g, gradient: 0% to 90% ethyl acetate in heptane) to afford the title compound as a pale yellow foam (152 mg, 74% yield). LCMS: m / z 428.3 [M+H] +, ESIpos。

[0627] Step C: 5-[5-Methyl-3-[[(3R)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenz ofuran-4-ol; dihydrochloride

[0628] To a solution of tert-butyl (3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidine-1-carboxylate (Example 9, Step B) (152 mg, 0.36 mmol, 1.0 eq) in CH2Cl2 (6 mL) and MeOH (2 mL) at +10 °C was added dropwise 4M HCl in 1,4-dioxane (1.07 g, 0.89 mL, 3.56 mmol, 10 eq). The reaction mixture was stirred at 23 °C for 16 h. The reaction mixture was evaporated to dryness to afford the title compound as a pale yellow foam (145 mg, 97% yield). LCMS: m / z 326.1 [M-H] - , ESI neg。

[0629] Step D: Methyl 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin- 3-yl]amino]-1-piperidinyl]butanoate

[0630] To a suspension of 5-[5-methyl-3-[[(3R)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; dihydrochloride (Example 9, Step C) (145 mg, 0.34 mmol, 1.0 eq) in dichloromethane (7 mL) at 0 °C was added methyl 4-oxobutanoate (CAS#13865-19-5, 111 mg, 0.10 mL, 0.86 mmol, 2.50 eq), followed by sodium acetate (CAS#13865-19-5, 111 mg, 0.10 mL, 0.86 mmol, 2.50 eq) and sodium triacetoxyborohydride (CAS#56553-60-7, 131 mg, 0.62 mmol, 1.80 eq). The reaction mixture was stirred at 0 °C for 5 min and at room temperature for 3 h. Saturated NaHCO3 solution (30 ml) was added to the reaction mixture and the mixture was extracted with dichloromethane (3 x 50 mL). The organic phase was separated and washed with water (20 ml) and brine (20 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash chromatography (silica gel, 12 g, gradient: 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound as a yellow foam (89 mg, 57% yield). MS: m / z 426.2 [M-H] - , ESIneg。

[0631] Step E: 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin- 3-yl]amino]-1-piperidinyl]butanoic acid

[0632] To a solution of methyl 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyrate (Example 9, Step D) (88 mg, 0.20 mmol, 1.0 eq) in tetrahydrofuran (1 mL) and methanol (0.5 mL) was added dropwise 1 M aqueous LiOH (0.59 mL, 0.59 mmol, 3.0 eq). The yellow reaction solution was stirred at 23 °C for 16 h. The reaction mixture was neutralized with 5% citric acid and adjusted to pH = 4. The aqueous phase was saturated with solid sodium chloride and extracted with dichloromethane (4 x 30 mL). The combined organic extracts were dried over sodium sulfate, filtered and the solvent was evaporated in vacuo to afford the title compound as a pale yellow solid (38 mg, 45% yield). MS: m / z 412.1 [M-H] - , ESI neg.

[0633] Example 10:

[0634] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl] amino]-1-piperidinyl]-N-methyl-butyramide

[0635]

[0636] To a solution of the aforementioned 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyric acid (Example 9, Step E) (6.4 mg, 0.015 mmol, 1.00 eq) and N,N-dimethylformamide (0.4 mL) was added N,N-diisopropylethylamine (5.7 mg, 0.008 mL, 0.044 mmol, 3.00 eq), followed by HATU (CAS# 148893-10-1, 7.30 mg, 0.019 mmol, 1.30 eq). The yellow solution was stirred for 2 min and then methylamine hydrochloride (CAS# 593-51-1, 1.50 mg, 0.022 mmol, 1.50 eq) was added. The reaction mixture was stirred at 23 °C for 60 h. The reaction mixture was quenched with 0.5 mL of water. The crude product was then purified by preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 x 30 mm; conditions: MeCN / water + 0.1% triethylamine; gradient: 20% to 80% MeCN in water, run time 4.5 min) to afford the title compound as a pale yellow solid (3.3 mg, 50% yield). LCMS: m / z 427.2 [M+H] + , ESI pos.

[0637] Example 11:

[0638] 5-[5-Methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidinyl]amino]-1,2,4-triazin-6- yl]-2,3-

[0639] dihydrobenzofuran-4-ol

[0640]

[0641] Step A: Methyl 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]propionate

[0642] To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidinyl]amine 1:2 hydrochloride (Example 1, Step E) (171 mg, 0.455 mmol, 1.0 eq) in dry tetrahydrofuran (2 mL) and dry N,N-dimethylformamide (2 mL) was added N,N-diisopropylethylamine (294 mg, 0.387 mL, 0.0023 mmol, 5.0 eq), followed by the dropwise addition of methyl 3-bromopropionate (CAS# 3395-91-3, 114 mg, 0.075 mL, 0.683 mmol, 1.5 eq). The reaction mixture was stirred at 60 °C for 20 h. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (2 x 40 mL). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (0% to 50% ethyl acetate in heptane; then ethyl acetate:methanol 9:1 (v / v)) to afford the title compound as a colorless oil (108 mg, 72% yield). LCMS: m / z 314.1 [M+H] + , ESIpos.

[0643] Step B3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]-N-(2,2- dimethoxyethyl)propanamide

[0644] A solution of methyl 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidino]propionate (Example 11, Step A) (108 mg, 0.33 mmol, 1.0 eq) in tetrahydrofuran (2 mL) and methanol (1 mL) was added dropwise to 1 M aqueous lithium hydroxide in dioxane (0.425 mL, 0.425 mmol, 1.3 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was then concentrated. The residue was dissolved twice in toluene, concentrated and dried at 50 °C under high vacuum for 1 h. The residue was dissolved in dry N,N-dimethylformamide (3 mL), and N-ethyldiisopropylamine (CAS# 7087-98-5, 211.3 mg, 0.278 mL, 1.6 mmol, 5.0 eq) and HATU (CAS# 148893-10-1, 187 mg, 0.49 mmol, 1.5 eq) were added at ambient temperature. After stirring for 2 min, 2,2-dimethoxyethylamine (62 mg, 0.064 mL, 0.589 mmol, 1.8 eq) was added. The reaction mixture was stirred at 23 °C for 3 h. The reaction mixture was quenched with water (15 mL) and extracted with ethyl acetate (2 x 30 mL). The organic layer was washed with brine (30 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (gradient: 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound as a light brown oil (137 mg, 97% yield). LCMS: m / z 387.3 [M+H] + , ESI pos.

[0645] Step C: 6-Chloro-5-methyl-N-[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidinyl]-1,2,4-triazin-3- amine

[0646] A mixture of 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidino]-N-(2,2-dimethoxyethyl)propanamide (Example 11, Step B) (137 mg, 0.32 mmol, 1.0 eq) and Eaton's reagent (CAS# 39394-84-8, 3.79 g, 2.5 mL, 16 mmol, 50 eq) was stirred at 100 °C overnight. The reaction mixture was cooled to room temperature and added dropwise onto saturated aqueous NaHCO3 (100 mL) (strong gas evolution; pH check: ca. 7) and extracted with dichloromethane (3 x 60 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (gradient: 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound as a colorless oil (34 mg, 31% yield). LCMS: m / z 323.2 [M+H] + , ESIpos.

[0647] Step D: 5-[5-Methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidinyl]amino]-1,2,4-tri azin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0648] A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidinyl]-amine (Example 11, Step C) (34 mg, 0.1 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dihydrobenzofuran-4-ol (Example 1, Step C) (39 mg, 0.15 mmol, 1.5 eq) and cesium carbonate (88 mg, 0.3 mmol, 3.0 eq) in 1,4-dioxane (1.6 mL) and water (0.4 mL) was left under argon and XPhos Pd g3 gt (12.7 mg, 0.015 mmol, 0.15 eq) was added. The reaction mixture was stirred in a sealed tube at 90 °C for 3 h. The reaction mixture was cooled to room temperature, quenched with water (20 mL) and saturated aqueous NH4Cl (20 mL) and extracted with ethyl acetate (2 x 40 mL). The organic layer was washed with brine (40 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (gradient: 0% to 80% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound as a light brown foam (24 mg, 54% yield). LCMS: m / z 423.3 [M+H] + , ESIpos.

[0649] Example 12:

[0650] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N,N-dimethyl-butyramide

[0651]

[0652] To a solution of 4-[(3R)-3-[[6-(4-Hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (Example 9, Step E) (43 mg, 0.094 mmol, 1.0 eq) and freshly dried N,N-dimethylformamide (0.3 mL) was added N,N-diisopropylethylamine (60.5 mg, 0.08 mL, 0.468 mmol, 5.0 eq), followed by the addition of HATU (CAS# 148893-10-1, 55.0 mg, 0.14 mmol, 1.5 eq). The yellow solution was stirred for 5 minutes, then dimethylamine hydrochloride (CAS# 506-89-2, 13.7 mg, 0.168 mmol, 1.8 eq) was added. The reaction mixture was stirred at 23 °C for 60 hours. The reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (2 x 20 mL). The organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (gradient: 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)). The product was then purified by preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 x 30 mm; conditions: ACN / water + 0.1% TEA; gradient: ACN in water, run time 4.5 min) to afford the title compound as a light brown foam (9 mg, 21% yield). LCMS: m / z 441.5 [M+H] + , ESI pos.

[0653] Example 13:

[0654] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl] amino]-1-piperidinyl]-1-pyrrolidin-1-yl-butan-1-one

[0655]

[0656] To a solution of 4-[(3R)-3-[[6-(4-hydroxy-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butanoic acid (Example 9, Step E) (30.0 mg, 0.065 mmol, 1.0 eq) and dry N,N-dimethylformamide (0.265 mL), N,N-diisopropylethylamine (42.2 mg, 0.056 mL, 0.327 mmol, 5.0 eq) was added, followed by HATU (CAS# 148893-10-1, 38.4 mg, 0.098 mmol, 1.5 eq). The yellow solution was stirred for 5 min and then pyrrolidine (CAS# 123-75-1, 8.4 mg, 0.0097 mL, 0.118 mmol, 1.8 eq) was added. The reaction mixture was stirred at 23 °C for 16 h. Then, HATU (38.4 mg, 0.098 mmol, 1.5 eq), N,N-diisopropylethylamine (42.2 mg, 0.059 mL, 0.327 mmol, 5.0 eq) and 4-[(3R)-3-[[6-(4-hydroxy-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butanoic acid (30 mg, 0.065 mmol, 1.0 eq) were added to the reaction mixture and stirring was continued at 23 °C for 48 h. The reaction mixture was quenched with 0.5 mL of water. The crude product was then purified by preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 x 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: ACN in water, run time 4.5 min) to afford the title compound as a yellow solid (7.3 mg, yield 22%). LCMS: m / z 467.4 [M+H] + , ESI pos.

[0657] Example 14:

[0658] 4-[(3R)-3-[[6-(4-4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl- 1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N-(2-hydroxyethyl)-N-methyl-butyramide

[0659]

[0660] To a solution of 4-[(3R)-3-[[6-(4-hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butanoic acid (Example 9, Step E) (30 mg, 0.065 mmol, 1.0 eq) and dry N,N-dimethylformamide (0.265 mL) was added N,N-diisopropylethylamine (42.2 mg, 0.056 mL, 0.327 mmol, 5.0 eq), followed by the addition of HATU (CAS# 148893-10-1, 38.4 mg, 0.098 mmol, 1.5 eq). The yellow solution was stirred for 2 min and then 2-(methylamino)ethanol (CAS# 109-83-1, 8.83 mg, 0.0094 mL, 0.118 mmol, 1.8 eq) was added. The reaction mixture was stirred at 23 °C for 16 h. HATU (38.4 mg, 0.098 mmol, 1.5 eq), N,N-diisopropylethylamine (42.2 mg, 0.0556 mL, 0.327 mmol, 5.0 eq) and 4-[(3R)-3-[[6-(4-hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butanoic acid (Example 9, Step E) (30 mg, 0.065 mmol, 1.0 eq) were added to the reaction mixture. The reaction mixture was quenched with 0.5 mL of water. The crude product was then purified by preparative HPLC (column: YMC-Triart C18, 12 nm, 5 um, 100 x 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: ACN in water, run time 4.5 min) to give the title compound as a yellow solid (6 mg, yield 18%). LCMS: m / z 471.4 [M+H] + , ESI pos.

[0661] Example 15:

[0662] 1-(3-Hydroxypyrrolidin-1-yl)-4-[racemic-(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran- 5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butan-1-one; formic acid

[0663]

[0664] To a solution of 4-[(3R)-3-[[6-(4-hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butanoic acid (Example 9, Step E) (34 mg, 0.074 mmol, 1.0 eq) and dry N,N-dimethylformamide (0.3 mL) was added N,N-diisopropylethylamine (47.8 mg, 0.063 uL, 0.37 mmol, 5.0 eq), followed by the addition of HATU (CAS# 148893-10-1, 43.5 mg, 0.111 mmol, 1.5 eq). The yellow solution was stirred for 2 min and then DL-pyrrolidin-3-ol (CAS# 40499-83-0, 11.6 mg, 0.011 mL, 0.133 mmol, 1.8 eq) was added. The reaction mixture was stirred at 23 °C for 60 h. The reaction mixture was quenched with 0.5 mL of water and concentrated. The product was purified by preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 x 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: ACN in water, run time 4.5 min) to afford the title compound as a light brown foam (11 mg, yield 27%). LCMS: m / z 483.5 [M+H] + , ESI pos.

[0665] Example 16:

[0666] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl] amino]-1-piperidinyl]-1-N-morpholinyl-butan-1-one

[0667]

[0668] To a solution of 4-[(3R)-3-[[6-(4-hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butanoic acid (Example 9, Step E) (40 mg, 0.097 mmol, 1.0 eq) and dry N,N-dimethylformamide (2.5 mL) was added N,N-diisopropylethylamine (62.5 mg, 0.082 mL, 0.484 μmol, 5.0 eq), followed by the addition of HATU (CAS# 148893-10-1, 55.2 mg, 0.145 mmol, 1.5 eq). The yellow solution was stirred for 4 min, then morpholine (CAS# 110-91-8, 15.2 mg, 0.015 mL, 0.174 mmol, 1.8 eq) was added. After 24 h, N,N-diisopropylethylamine (62.5 mg, 0.082 mL, 0.484 mmol, 5.0 eq) and HATU (55.2 mg, 0.145 mmol, 1.5 eq) were added again. After stirring for 4 min, morpholine (15.2 mg, 0.015 mL, 0.174 mmol, 1.8 eq) was added again. The reaction mixture was stirred at 23 °C for a total of 45 h. The reaction mixture was quenched with 2 mL of water. The reaction mixture was extracted with water (ca. 10 mL) and ethyl acetate (ca. 15 mL), and then the aqueous layer was back-extracted with ethyl acetate (ca. 15 mL). The organic layer was washed with brine (ca. 10 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The product was then purified by preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 x 30 mm; conditions: ACN / water + 0.1% TEA; gradient: ACN in water, run time 4.5 min) to afford the title compound as a pale yellow lyophilized solid (4 mg, yield 8%). LCMS: m / z 483.5 [M+H] + , ESI pos.

[0669] Example 17:

[0670] 5-[5-Methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidinyl]amino]-1,2,4-triazin-6- yl]-2,3-dihydrobenzofuran-4-ol

[0671]

[0672] Step A: 6-Chloro-5-methyl-N-[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidinyl]-1,2,4-triazin-3- amine

[0673] To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-piperidin-3-yl]amine (Example 1, Step E) (250 mg, 1.1 mmol, 1.0 eq) in dry tetrahydrofuran (4 mL) and dry N,N-dimethylformamide (4 mL) was added N,N-diisopropylethylamine (568 mg, 0.7467 mL, 4.4 mmol, 4.0 eq), followed by the addition of 1-(2-bromoethyl)pyrazole; hydrobromide (CAS# 1955531-53-9, 443.7 mg, 1.65 mmol, 1.5 eq). The reaction mixture was stirred at 60 °C for 20 h. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2 x 60 mL). The organic layer was washed with brine (60 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (0% to 100% ethyl acetate in heptane) to afford the title compound as a light brown oil (231 mg, 62% yield). LCMS: m / z 322.3 ([{35Cl}M+H] + ), 324.2 ([{37Cl}M+H] + ), ESIpos.

[0674] Step B: 5-[5-Methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidinyl]amino]-1,2,4-tri azin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0675] A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(pyrazol-1-ylethyl)-3-piperidinyl]-amine (Example 17, Step A) (113 mg, 0.351 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dihydrobenzofuran-4-ol (Example 1, Step C) (128.9 mg, 0.492 mmol, 1.4 eq) and cesium carbonate (343 mg, 1.05 mmol, 3.0 eq) in 1,4-dioxane (4 mL) and water (1 mL) was left under argon. Finally, XPhos Pd g3 gt (44.6 mg, 0.053 mmol, 0.15 eq) was added. The reaction mixture was stirred in a sealed tube at 90 °C for 3 h. The reaction mixture was cooled to room temperature and quenched with water (20 mL) and saturated aqueous NH4Cl solution (20 mL), then extracted with ethyl acetate (2 x 40 mL). The organic layer was washed with brine (40 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (gradient: 0% to 80% (dichloromethane:methanol:NH4OH 110:10:1) in dichloromethane) to afford the title compound as a yellow foam (89 mg, 57% yield). LCMS: m / z 422.4 [M+H]+ , ESIpos.

[0676] Examples 18 and 19:

[0677] 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-tri azin-6-yl]-2,3-dihydrobenzofuran-4-ol; formic acid and

[0678] 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl] amino]-1-piperidinyl]butanenitrile

[0679]

[0680] Step A: 4-[(3R)-3-[(6-Chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]butanenitrile

[0681] To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidinyl]amine (Example 1, Step E) (400 mg, 1.76 mmol, 1.0 eq) in dry tetrahydrofuran (6 mL) and dry N,N-dimethylformamide (6 mL) was added N,N-diisopropylethylamine (567 mg, 0.747 mL, 4.4 mmol, 2.5 eq), followed by the dropwise addition of 4-bromobutyronitrile (CAS# 5332-06-9, 390 mg, 0.26 mL, 2.64 mmol, 1.5 eq). The reaction mixture was stirred at 60 °C for 16 h. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2 x 60 mL). The organic layer was washed with brine (60 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (0% to 100% ethyl acetate in heptane) to afford the title compound as a light brown oil (391 mg, 72% yield). LCMS: m / z 295.2 ([{35Cl}M+H] + ), 297.2 ([{37Cl}M+H] + ), ESIpos.

[0682] Step B: 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin- 3-yl]amino]-1-piperidinyl]butanenitrile

[0683] 4-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidino]butyronitrile (Example 18, Step A) (150 mg, 0.51 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dihydrobenzofuran-4-ol (Example 1, Step C) (187 mg, 0.712 mmol, 1.4 eq) and cesium carbonate (498 mg, 1.53 mmol, 3.0 eq) in a mixture of 1,4-dioxane (4 mL) and water (1 mL) were left under argon and finally XPhos Pd g3 gt (65 mg, 0.076 mmol, 0.15 eq) was added. The reaction mixture was stirred in a sealed tube at 90 °C for 3 h. The reaction mixture was cooled to room temperature and quenched with water (20 mL) and saturated aqueous NH4Cl solution (20 mL), then extracted with ethyl acetate (2 x 40 mL). The organic layer was washed with brine (40 mL), dried over sodium sulfate, filtered off and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (gradient: 0% to 80% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)), followed by a second purification by flash chromatography (SiO2-amine; 0% to 100% ethyl acetate in heptane) to give the title compound as a yellow foam (145 mg, 69% yield). LCMS: m / z 395.2 [M+H] + , ESIpos.

[0684] Step C: 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2, 4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formic acid

[0685] A solution of 4-[(3R)-3-[[6-(4-hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-1-yl]butyronitrile (Example 18, Step B) (135 mg, 0.343 mmol, 1.0 eq) in dry N,N-dimethylformamide (1.45 mL) was added sodium azide (CAS# 26628-22-8, 100 mg, 1.54 mmol, 4.5 eq), followed by L-proline (CAS# 147-85-3, 11.8 mg, 0.103 mmol, 0.3 eq). The reaction mixture was stirred in a sealed tube at 115 °C for 16 h. After 16 h, more sodium azide (100 mg, 1.54 mmol, 4.5 eq) and L-proline (11.8 mg, 0.103 mmol, 0.3 eq) were added and stirring was continued at 120 °C for 60 h. The reaction mixture was quenched with water (5 mL) and extracted with ethyl acetate (2 × 30 mL). The aqueous phase was evaporated, the solid residue was triturated in DCM / MeOH 9:1, some sodium sulfate was added, and then filtered off. The product was then purified by preparative HPLC (column: YMC-Triart C18, 12 nm, 5 μm, 100 x 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: 5% to 50% ACN, run time 4.5 min) to give the title compound as a light brown lyophilized solid (51 mg, yield 29%). LCMS: m / z 438.4 [M+H] + , ESI pos.

[0686] Example 20:

[0687] 5-[5-Methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidinyl]amino]-1,2,4-triazin-6- yl]-2,3-

[0688] dihydrobenzofuran-4-ol

[0689]

[0690] Step A: 6-Chloro-5-methyl-N-[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidinyl]-1,2,4-triazin-3- amine

[0691] At 0 °C, a solution of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-piperidin-3-yl]amine 1:2 hydrochloride (Example 1, Step B) (67 mg, 0.178 mmol, 1.0 eq) in dry dichloromethane (2 mL) was added 3-oxazol-2-ylpropanal (CAS# 1214937-88-8, 31.2 mg, 0.25 mmol, 1.4 eq), followed by the addition of sodium acetate (36.6 mg, 0.446 mmol, 2.0 eq) and sodium triacetoxyborohydride (CAS# 56553-60-7, 68 mg, 0.321 mmol, 1.8 eq). The reaction mixture was stirred at 0 °C for 5 min and at room temperature for 3 h. Saturated NaHCO3 solution (20 mL) was added to the reaction mixture and extracted with dichloromethane (3 x 30 mL). The organic phase was separated and washed with water (20 mL) and brine (20 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (gradient: 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound as a light brown oil (43 mg, yield 72%). LCMS: m / z 337.2 ([{35Cl}M+H] + ), 339.2 ([{37Cl}M+H] + ), ESIpos.

[0692] Step B: 5-[5-Methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidinyl]amino]-1,2,4-tri azin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0693] (6-Chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidinyl]amine (Example 20, Step A) (43 mg, 0.128 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dihydrobenzofuran-4-ol (Example 1, Step C) (55.8 mg, 0.192 mmol, 1.5 eq), and cesium carbonate (125 mg, 0.383 mmol, 3.0 eq) in a mixture of 1,4-dioxane (1.6 mL) and water (0.4 mL) were left standing under argon, and finally XPhos Pd G3 gt (16 mg, 0.019 mmol, 0.15 eq) was added. The reaction mixture was stirred in a sealed tube at 90 °C for 2 h. The reaction mixture was cooled to room temperature and quenched with water (10 mL) and saturated aqueous NH4Cl solution (10 mL), and then extracted with ethyl acetate (2 x 20 mL). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography on silica gel (gradient: 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to give the title compound as a pale yellow foam (19 mg, 32% yield). LCMS: m / z 437.3 [M+H] + , ESI pos.

[0694] Example 21:

[0695] 5-[5-Methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidinyl]amino]-1,2,4-tri azin-6-

[0696] yl]-2,3-dihydrobenzofuran-4-ol

[0697]

[0698] Step A: 3-[(3R)-3-[(6-Chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidinyl]propanenitrile

[0699] To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-piperidin-3-yl]amine (Example 1, Step E) (500 mg, 2.2 mmol, 1.0 eq) in dry tetrahydrofuran (8 mL) and dry N,N-dimethylformamide (8 mL) was added N,N-diisopropylethylamine (710 mg, 0.934 mL, 5.5 mmol, 2.5 eq), followed by the dropwise addition of 3-bromopropionitrile (CAS# 2417-90-5, 465 mg, 0.287 mL, 3.3 mmol, 1.5 eq). The reaction mixture was stirred at 60 °C for 48 h. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2 x 60 mL). The organic layer was washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (0% to 100% ethyl acetate in heptane) to afford the title compound as a light brown oil (425 mg, 65% yield). LCMS: m / z 281.2 ([{35Cl}M+H] + ), 283.2 ([{37Cl}M+H] + ), ESI pos.

[0700] Step B: 3-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin- 3-yl]amino]-1-piperidinyl]propanenitrile

[0701] A mixture of 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidin-1-yl]propanenitrile (Example 22, Step A) (200 mg, 0.712 mmol, 1.0 eq) was dissolved in dry 1,4-dioxane (4 mL) and water (1 mL). Then, under an argon atmosphere, 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dihydrobenzofuran-4-ol (Example 1, Step C) (280 mg, 1.07 mmol, 1.5 eq), cesium carbonate (696 mg, 2.2 mmol, 3.0 eq), and XPhos PdG3 (90.5 mg, 0.10 mmol, 0.15 eq) were added. The reaction mixture was sealed and stirred at 100 °C for 1.5 h. The reaction mixture was quenched with water (10 mL) and saturated aqueous NH4Cl (10 mL), then extracted with ethyl acetate (2 x 25 mL). The organic layer was washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel (gradient: 0% to 80% (dichloromethane:methanol:NH4OH 110:10:1) in dichloromethane) to afford the title compound as a light brown oil (240 mg, 80% yield). LCMS: m / z 381.3 [M+H] + , ESI pos.

[0702] Step C: 5-[5-Methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidinyl]amino]-1,2, 4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol

[0703] To a solution of 3-[(3R)-3-[[6-(4-hydroxydihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]propanenitrile (Example 22, Step B) (100 mg, 0.237 mmol, 1.0 eq) in dry N,N-dimethylformamide (1 mL) was added L-proline (CAS# 147-85-3, 8.2 mg, 0.071 μmol, 0.30 eq), followed by the addition of sodium azide (CAS# 26628-22-8, 69 mg, 1.06 mmol, 4.5 eq). The reaction mixture was stirred at 115 °C for 16 h. After 16 h, L-proline (8.2 mg, 0.071 mmol, 0.3 eq) and sodium azide (69 mg, 1.06 mmol, 4.5 eq) were added again. The reaction mixture was stirred at 115 °C for 48 h. The reaction mixture was quenched with water (ca. 5 mL) and then extracted with ethyl acetate (ca. 25 mL). The product was in the aqueous phase. The aqueous layer was back-extracted four times with ethyl acetate (ca. 25 mL). The aqueous layer was concentrated in vacuo. Then the crude product was purified by preparative HPLC (column: YMC-Triart C 18 , 12 nm, 5 μm, 100 x 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: 5% to 50% ACN, run time 4.5) to give the title as a light brown lyophilized solid (3 mg, yield 3%). LCMS: m / z 242.3 [M+H] + , ESI pos.

[0704] Example 22:

[0705] 5-[5-Methyl-3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidinyl]amino]-1,2,4-triazin-6- yl]-2,3-dihydrobenzofuran-4-ol

[0706]

[0707] Step A: 6-Chloro-5-methyl-N-[(3R)-1-(3-methylsulfonylpropyl)-3-piperidinyl]-1,2,4-triazin- 3-amine

[0708] To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-piperidin-3-yl]amine (Example 1, Step E) (100 mg, 0.439 mmol, 1.0 eq) in dry tetrahydrofuran (1.6 mL) and dry N,N-dimethylformamide (1.6 mL) was added N,N-diisopropylethylamine (142.0 mg, 0.187 mL, 1.1 mmol, 2.5 eq), followed by the dropwise addition of 1-bromo-3-methylsulfonyl-propane (CAS# 859940-73-1, 132.5 mg, 0.659 mmol, 1.5 eq). The reaction mixture was stirred at 60 °C for 12 h. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (2 x 40 mL). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, 12 g; 0% to 10% MeOH in DCM) to afford the title compound as an orange oil (111 mg, 65% yield). LCMS: m / z 348.2 [M+H] + , ESIpos.

[0709] Step B: 5-[3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidinyl]amino]-5-methyl-1,2,4-tri azin-6-yl]dihydrobenzofuran-4-ol

[0710] A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(3-methylsulfonylpropyl)piperidin-3-yl]amine (Example 23, Step A) (108 mg, 0.310 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dihydrobenzofuran-4-ol (Example 1, Step C) (144.67 mg, 0.497 mmol, 1.6 eq), cesium carbonate (305 mg, 0.936 mmol, 3.0 eq) and XPhos Pd g3 (39.4 mg, 0.047 mmol, 0.15 eq) in 1,4-dioxane (1.9 mL) and water (0.46 mL) was flushed with argon and stirred at 90 °C for 3 h. The reaction mixture was cooled to room temperature and extracted with ethyl acetate and semi-saturated NH4Cl solution. The aqueous layer was back-extracted with ethyl acetate. The organic layers were washed with water and brine. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash chromatography (Si-amine, 12 g, gradient: 0% to 10% methanol in DCM) to afford the title compound as a yellow solid (108 mg, 70% yield). LCMS: m / z 448.3 [M+H] + , ESIpos.

[0711] Example A

[0712] The compound of formula I can be used, in a manner known per se, as an active ingredient for producing tablets having the following composition:

[0713]

[0714] Example B

[0715] The compound of formula I can be used, in a manner known per se, as an active ingredient for producing capsules having the following composition:

[0716]

Claims

1. A compound of formula I wherein R 1 and R 5 forms with the atom to which it is bonded i. a 4- to 6-membered heterocycle containing a single O heteroatom, or ii. a 4- to 5-membered cycloalkyl ring; R 2 and R 3 is selected from H and alkyl, where R 2 or R 3 only one of which may be H or alkyl; R 4 Selected from i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkyl substituted with hydroxy or alkoxy, v. cycloalkylalkyl substituted with hydroxy, alkoxy or –COOH, vi. a 4- to 6-membered heterocycle containing a single O heteroatom, vii. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 2 or 3, viii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is a tetrazole, oxadiazole or oxazole heteroaryl, ix. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; x.-(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3, xi.-(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3 R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form a. a 5-membered heterocycle, wherein the heterocycle is optionally substituted with OH, or b. a 6-membered heterocycle additionally containing 1 O heteroatom; R 6 is H or –OH, Wherein when R 6 is –OH, R 4 can only be an alkyl group; and pharmaceutically acceptable salts.

2. The compound according to claim 1, wherein R 1 and R 5 form a 5-membered heterocycle containing a single O heteroatom with the atom to which it is bonded.

3. The compound according to claim 1 or claim 2, wherein R 2 is H and R 3 is alkyl.

4. The compound according to any one of claims 1 to 3, wherein R 4 is selected from i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy, v. a 4-membered heterocycle containing a single O heteroatom, vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 3, vii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is an oxazole heteroaryl, viii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; ix.-(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3, x.-(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3, R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form a. a 5-membered heterocycle, wherein the heterocycle is optionally substituted with OH, or b. a 6-membered heterocycle additionally containing 1 O heteroatom; Wherein when R 6 is –OH, R 4 can only be an alkyl group.

5. The compound according to any one of claims 1 to 3, wherein R 4 is selected from i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy, v. a 4-membered heterocycle containing a single O heteroatom, vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, wherein n is 3, vii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is an oxazole heteroaryl; Wherein when R 6 is –OH, R 4 can only be an alkyl group.

6. The compound according to any one of claims 1 to 3, wherein R 4 is selected from i. cycloalkyl substituted with hydroxy or alkoxy, ii. cycloalkylalkyl substituted with hydroxy or alkoxy, and iii. a 4- to 6-membered heterocycle containing a single O heteroatom.

7. A compound according to any one of claims 1 to 3, wherein R 4 is selected from i. cycloalkyl substituted with hydroxy or alkoxy, and ii. cycloalkylalkyl substituted with hydroxy or alkoxy.

8. The compound according to any one of claims 1 to 7, wherein R 6 is H.

9. The compound according to claim 1, wherein R 1 and R 5 forms a 5-membered heterocycle containing a single O heteroatom with the atom to which it is bonded; R 2 is H and R 3 is methyl; R 4 selected from i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy, v. a 4-membered heterocycle containing a single O heteroatom, vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, where n is 3, vii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is an oxazole heteroaryl, viii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole, ix.-(CH2) n S(O)2CH3 or (CH2) n CN, where n is 3 x.-(CH2) n C(O)NR'R", where n is 3 and both R' and R" are CH3, R' is CH3 and R" is hydroxyalkyl, or R' and R" together with the N to which they are attached form a. a 5-membered heterocycle, wherein the heterocycle is optionally substituted with OH, or b. a 6-membered heterocycle additionally containing 1 O heteroatom; R 6 is H or –OH, Wherein when R 6 is –OH, R 4 can only be an alkyl group; and pharmaceutically acceptable salts.

10. The compound according to claim 1, wherein R 1 and R 5 forms a 5-membered heterocycle containing a single O heteroatom with the atom to which it is bonded; R 2 is H and R 3 is methyl; R 4 selected from i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy, v. a 4-membered heterocycle containing a single O heteroatom, vi. (CH2) n C(O)OH, (CH2) n C(O)OCH3 or (CH2) n C(O)NHCH3, wherein n is 3, vii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is an oxazole heteroaryl; R 6 is H or --OH, Wherein when R 6 is –OH, R 4 can only be an alkyl group; and pharmaceutically acceptable salts.

11. The compound according to claim 1, wherein the compound is 5-[3-[[(3R)-1-(2-hydroxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol or a pharmaceutically acceptable salt thereof.

12. The compound according to claim 1, wherein the compound is selected from 5-[5-methyl-3-[[(±)-(3R)-1-(2-hydroxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formate; 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidinyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; (3S,5R)-1-ethyl-5-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidin-3-ol; 5-[5-methyl-3-[[(±)-(3R)-1-(2-methoxypropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyric acid; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]methyl butyrate; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N-methyl-butyramide; 5-[5-Methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; and its pharmaceutically acceptable salts.

13. The compound according to claim 1, wherein the compound is selected from 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N,N-dimethyl-butyramide; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-1-pyrrolidin-1-yl-butan-1-one; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N-(2-hydroxyethyl)-N-methyl-butyramide; 1-(3-Hydroxypyrrolidin-1-yl)-4-[racemic-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butan-1-one; formic acid; 1-(3-Hydroxypyrrolidin-1-yl)-4-[racemic-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butan-1-one; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-1-N-morpholinyl-butan-1-one; 5-[5-Methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formic acid; 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butanenitrile; 5-[5-Methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N,N-dimethyl-butyramide; formic acid" 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N,N-dimethyl-butyramide; formic acid; 5-[5-Methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-Methyl-3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; and its pharmaceutically acceptable salts.

14. The compound according to claim 1, wherein the compound is 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]butyric acid, or a pharmaceutically acceptable salt thereof.

15. The compound according to claim 1, wherein the compound is selected from 5-[5-Methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidinyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-N,N-dimethyl-butyramide; 4-[(3R)-3-[[6-(4-Hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidinyl]-1-pyrrolidin-1-yl-butan-1-one; and its pharmaceutically acceptable salts.

16. The compound according to any one of claims 1 to 15, which is used as a therapeutic active substance.

17. The compound according to any one of claims 1 to 15, which is used for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

18. A pharmaceutical composition, which comprises the compound according to any one of claims 1 to 15 and a therapeutically inert carrier.

19. Use of the compound according to any one of claims 1 to 15 for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

20. The compound according to any one of claims 1 to 15, which is used for treating or preventing cardiovascular diseases, disorders or conditions.

21. Use of the compound according to any one of claims 1 to 15 in the treatment or prevention of cardiovascular diseases, disorders or conditions. Use of a compound according to any one of claims 1 to 15 for the preparation of a medicament for the treatment or prevention of a cardiovascular disease, disorder or condition.

23. A method of inhibiting NLRP3, the method comprising administering an effective amount of a compound as claimed in any one of claims 1 to 15 to inhibit NLRP3.

24. A method of treating or preventing a cardiovascular disease, disorder or condition, the method comprising administering an effective amount of a compound according to any one of claims 1 to 15.

25. The present invention as described in the specification.

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