Novel compounds as modulators of NLRP3 inhibition

Abstract

The present invention relates to a compound represented by general formula (I) TIFF2025540202000038.tif30161 (In the formula, R 1 , R 2 , R 3 , R 4 , R 5 and R 6 The present invention relates to novel compounds having the formula (I) as described herein, compositions comprising the compounds, and methods of using the compounds.

Description

[Technical Field]

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

[0002] The present invention relates to novel compounds of formula I, [ka] During the ceremony, R 1 and R 5 , and the atoms to which they are attached, i. a 4- to 6-membered heterocycle containing a single O heteroatom, or ii. Forms any of 4- to 5-membered cycloalkyl rings; R 2 and R 3 is selected from H and alkyl, and R 2 or R 3 Only one of may be H or alkyl; R 4 is selected from: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkyl substituted with hydroxy or alkoxy; v. Cycloalkylalkyl substituted with hydroxyl, alkoxy or -COOH; vi. 4- to 6-membered heterocycles containing a single O heteroatom; vii.(CH2) n C(O)OH, (CH2) n C(O)OCH3, or (CH2) n C(O)NHCH3 (wherein n is 2 or 3), viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is 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 (wherein n is 3), xi.-(CH2) n C(O)NR'R'' where n is 3 and R' and R'' are both CH3, or R' is CH3 and R'' is hydroxylalkyl, or R' and R'' together with the N to which they are attached are a. a 5-membered heterocycle, the heterocycle being optionally substituted with OH; or b. Forming any six-membered heterocycle further containing one O heteroatom); R 6 is H or -OH; R 4 is R 6 may be alkyl only if -OH; Compounds and pharmaceutically acceptable salts are provided.

[0003] Furthermore, the present invention includes all racemic mixtures, all their corresponding enantiomers and / or optical isomers. [Background technology]

[0004] The NOD-like receptor (NLR) family pyrin domain-containing protein 3 (NLRP3) inflammasome is a component of inflammatory processes, and its aberrant activity is pathogenic in genetic disorders such as cryopyrin-associated periodic syndromes (CAPS), as well as complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer's disease, and atherosclerosis.

[0005] NLRP3 is an intracellular signaling molecule that senses many pathogen-, environmental, and host-derived factors. Upon activation, NLRP3 binds to apoptosis-associated speck-like proteins containing a caspase activation and recruitment domain (ASC). ASC then polymerizes to form large aggregates known as ASC specks. Polymerized 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 (pro-IL-1β and pro-IL-18, respectively), thereby activating these cytokines. Caspase-1 also mediates a type of inflammatory cell death known as pyroptosis. ASC specks can also recruit and activate caspase-8, which can process pro-IL-1β and pro-IL-18, leading to apoptotic cell death.

[0006] Caspase-1 cleaves pro-IL-1β and pro-IL-18 into their active forms, which are secreted from cells. Active caspase-1 also cleaves gasdermin-D, causing pyroptosis. Through its control of the pyroptotic cell death pathway, caspase-1 also mediates the release of alarmin molecules such as IL-33 and high-mobility group box 1 (HMGB1). Caspase-1 also cleaves intracellular IL-1R2, leading to its degradation and allowing IL-1α to be released. In human cells, caspase-1 can also regulate the processing and secretion of IL-37. Many other caspase-1 substrates, such as components of the cytoskeleton and glycolytic pathways, can contribute to caspase-1-dependent inflammation.

[0007] NLRP3-dependent ASC specks are released into the extracellular milieu where they can activate caspase-1, induce the processing of caspase-1 substrates, and propagate inflammation.

[0008] Active cytokines derived from NLRP3 inflammasome activation are key drivers of inflammation and interact with other cytokine pathways to shape immune responses 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 synergize with IL-23 to induce IL-17 production by memory CD4 Th17 cells and γδ T cells in the absence of T cell receptor engagement. IL-18 and IL-12 also act synergistically to induce IFN-γ production from memory T cells and NK cells, driving Th1 responses.

[0009] The inherited CAPS disorders Muckle-Wells syndrome (MWS), familial common cold autoinflammatory syndrome (FCAS), and neonatal-onset multisystem inflammatory disease (NOMID) are caused by gain-of-function mutations in NLRP3, defining it as a critical component of the inflammatory process. NLRP3 is also involved in the pathogenesis of many complex diseases, including metabolic disorders such as type 2 diabetes, atherosclerosis, obesity, and gout, among others.

[0010] The role of NLRP3 in central nervous system diseases is becoming clear, and lung diseases have also been shown to be affected by NLRP3. NLRP3 has also been suggested to play a role in several central nervous system conditions, including Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, and brain damage from pneumococcal meningitis (Walsh et al., Nature Reviews, 15:84-97, 2014, and Dempsey et al., Brain. Behav. Immun. 201761:306-316). NLRP3 has also been shown to play a role in many lung diseases, including chronic obstructive pulmonary disorder (COPD), asthma (including steroid-resistant asthma), asbestosis, and silicosis (De Nardo et al., Am. J. Pathol., 184:42-54, 2014 and Kim et al. Am. J Respir Crit Care Med. 2017 196(3):283-97). Furthermore, NLRP3 plays a role in the development of liver disease, kidney disease, and aging. Many of these associations are due to the involvement of NLRP3. - / - Although defined in mice, insights into the specific activation of NLRP3 in these diseases also exist. In type 2 diabetes (T2D), deposition of islet amyloid polypeptide in the pancreas activates NLRP3 and IL-1β signaling, leading to cell death and inflammation.

[0011] Several small molecules have been shown to inhibit the NLRP3 inflammasome. Glyburide inhibits IL-1β production at micromolar concentrations in response to NLRP3 activation, but not NLRC4 or NLRP1 activation. Other previously characterized weak NLRP3 inhibitors include parthenolide, 3,4-methylenedioxy-β-nitrostyrene, and dimethyl sulfoxide (DMSO), but these agents have limited potency and are nonspecific.

[0012] Current treatments for NLRP3-related diseases include biologic agents that target IL-1. These include the recombinant IL-1 receptor antagonist anakinra, the neutralizing IL-1β antibody canakinumab, and the soluble decoy IL-1 receptor rilonacept. These approaches have proven successful in treating CAPS, and these biologic agents are being used in clinical trials for other IL-1β-related diseases.

[0013] There is a need to provide compounds with improved pharmacological and / or physiological and / or physicochemical properties, and / or compounds that provide useful alternatives to known compounds. Furthermore, when developing NLRP3 inhibitors for treating peripheral indications, it is advantageous to minimize the exposure of NLRP3 inhibitor compounds in the brain compared to systemic exposure in order to minimize the risk of potential side effects in the central nervous system (CNS). Compounds of Formula I achieve this by demonstrating increased efflux and / or reduced permeability in transcellular assays expressing active P-gp protein. P-gp (P-glycoprotein) is an important transporter expressed in capillary endothelial cells that constitute the blood-brain barrier and blood-testis barrier, transporting xenobiotics back into the capillaries and limiting their exposure to the brain. Summary of the Invention

[0014] The present invention relates to novel compounds of formula I, [ka] During the ceremony, R 1 and R 5 , and the atoms to which they are attached, i. a 4- to 6-membered heterocycle containing a single O heteroatom, or ii. Forms any of 4- to 5-membered cycloalkyl rings; R 2 and R 3 is selected from H and alkyl, and R 2 or R 3Only one of may be H or alkyl; R 4 is selected from: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkyl substituted with hydroxy or alkoxy; v. Cycloalkylalkyl substituted with hydroxyl, alkoxy or -COOH; vi. 4- to 6-membered heterocycles containing a single O heteroatom; vii.(CH2) n C(O)OH, (CH2) n C(O)OCH3, or (CH2) n C(O)NHCH3 (wherein n is 2 or 3), viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is 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 (wherein n is 3), xi.-(CH2) n C(O)NR'R'' where n is 3 and R' and R'' are both CH3, or R' is CH3 and R'' is hydroxylalkyl, or R' and R'' together with the N to which they are attached are a. a 5-membered heterocycle, the heterocycle being optionally substituted with OH; or b. Forming any six-membered heterocycle further containing one O heteroatom); R 6 is H or -OH, R 4 is R 6 may be alkyl only if -OH; Compounds and pharmaceutically acceptable salts are provided. [Brief explanation of the drawings]

[0015] [Figure 1] The effects of compounds on hERG K+ current parameters will be evaluated at two concentrations in at least four cells. hERG studies will be performed using an automated patch clamp system, SynchroPatch® 384 (Nanion Technologies GmbH, Germany). K+ currents will be measured using the patch voltage clamp technique in the whole-cell configuration at 35-37°C. Cells will be held at a resting voltage of -80 mV and stimulated with the voltage pattern shown in Figure 1 (the pulse pattern used to elicit outward K+ currents at 35-37°C) at a stimulation frequency of 0.1 Hz (6 bpm) to activate hERG channels and conduct outward IKhERG currents.

[0016] The term "alkyl" refers to a monovalent linear or branched saturated hydrocarbon group of 1 to 6 carbon atoms. In some embodiments, unless otherwise specified, alkyl refers to a group having 1 to 6 carbon atoms (C 1~6 -alkyl) or 1 to 4 carbon atoms (C 1~4 -alkyl). 1~6 Examples of alkyl include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and pentyl. Particular alkyl groups include methyl and ethyl. Another particular alkyl group is propyl.

[0017] The term "alkoxy" refers to a group in which R' is C 1~6 represents a group of formula -O-R', which is an alkyl group. 1~6 Examples of -alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy.

[0018] The term "alkoxyalkyl" refers to an alkyl group in which one of the alkyl group's hydrogen atoms has been replaced by an alkoxy group. Examples of alkoxyalkyl include methoxymethyl and methoxyethyl. Specific examples of alkoxyalkyl include methoxyethyl and methoxypropyl.

[0019] The term "cycloalkyl" refers to a monocyclic or polycyclic saturated or partially unsaturated non-aromatic hydrocarbon. In some embodiments, unless otherwise specified, a cycloalkyl contains 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. In some embodiments, a cycloalkyl is a saturated monocyclic or polycyclic hydrocarbon. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0020] The term "cycloalkylalkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a cycloalkyl group. Examples of cycloalkylalkyl include cyclopropylmethyl and cyclobutylmethyl. A particular cycloalkylalkyl is cyclopropylmethyl.

[0021] The term "cyano" refers to the group -C≡N.

[0022] The term "heterocycle" refers to a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of 4 to 9 ring atoms containing one, two, or three ring heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Examples of monocyclic saturated heterocycles include azetidinyl, diazepanyl, pyrrolidinyl, tetrahydrofuranyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, and piperazinyl. Examples of polycyclic saturated heterocycles include azaspiroheptanyl, diazaspiroheptanyl, azaspirooctanyl, diazospirooctanyl, diazaspirononanyl, oxaazaspirooctanyl, and oxadiazaspirononanyl. A specific example of a heterocycle is piperidinyl. Another illustrative example of a heterocycle is oxetanyl.

[0023] The term "heteroaryl," alone or in combination, refers to a monovalent aromatic heterocyclic monocyclic or bicyclic ring system of 5 to 12 ring atoms containing 1, 2, 3, or 4 heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon. Examples of heteroaryl groups include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridinyl, pyridazinyl, pyrimidinyl, triazinyl, azepinyl, diazepinyl, isoxazolyl, benzofuranyl, isothiazolyl, benzothienyl, indolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzoxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalinyl, and benzothiophenyl. A specific example of a heteroaryl group is oxazole.

[0024] The term "heteroarylalkyl" refers to an alkyl group in which one of the alkyl group's hydrogen atoms is replaced by a heteroaryl group. Examples are oxyzolylethyl, tetrazolylethyl, and oxadiazolylethyl. A specific example is oxazolylethyl. Other examples are pyrazolylethyl, tetrazolylpropyl, oxazolylpropyl, and tetrazolylethyl.

[0025] The terms "hydroxy" or "hydroxyl" refer to an --OH group.

[0026] The term "hydroxyalkyl" refers to 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.

[0027] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of the free base or free acid and is not biologically or otherwise undesirable. 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, and N-acetylcysteine. In addition, these salts can be prepared by adding inorganic or organic bases to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, and polyamine resins. The compound of Formula I can also exist in the form of a zwitterion. Particularly preferred pharmaceutically acceptable salts of the compound of Formula I are salts formed with formic acid and with hydrochloric acid to produce the hydrochloride, dihydrochloride, or trihydrochloride salt.

[0028] The abbreviation uM means micromolar and is equivalent to the symbol μM.

[0029] The abbreviation uL means microliter and is equivalent to the symbol μL.

[0030] The abbreviation ug stands for microgram and is equivalent to the symbol μg.

[0031] The compounds of formula I may contain several asymmetric centers and may exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereomeric racemates or mixtures of diastereomeric racemates.

[0032] According to the Cahn-Ingold-Prelog rules, the asymmetric carbon atom can be of the "R" or "S" configuration.

[0033] Also, one embodiment of the present invention provides compounds according to formula I as described herein and pharmaceutically acceptable salts or esters thereof, particularly compounds according to formula I as described herein and pharmaceutically acceptable salts thereof, more particularly compounds according to formula I as described herein.

[0034] One embodiment of the present invention is R 1 and R 5 and the atoms to which they are attached form a 4-6 membered heterocycle containing a single O heteroatom, according to formula I as described herein.

[0035] One embodiment of the present invention is R 1 and R 5 and the atoms to which they are attached form a 4-5 membered heterocycle containing a single O heteroatom, according to formula I as described herein.

[0036] One embodiment of the present invention is R 1 and R 5 and the atoms to which they are attached form a 5-membered heterocycle containing a single O heteroatom.

[0037] One embodiment of the present invention is R 2 is H and R 3 is alkyl.

[0038] One embodiment of the present invention is R2 is H and R 3 is methyl.

[0039] One embodiment of the present invention is R 4 is selected from the following: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkyl substituted with hydroxy or alkoxy; v. Cycloalkylalkyl substituted with hydroxyl, alkoxy or -COOH; vi. 4- to 6-membered heterocycles containing a single O heteroatom; vii.(CH2) n C(O)OH, (CH2) n C(O)OCH3, or (CH2) n C(O)NHCH3 (wherein n is 2 or 3), viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazoleheteroaryl; ix. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; x.-(CH2) n S(O)2CH3 or (CH2) n CN (wherein n is 3), xi.-(CH2) n C(O)NR'R'' where n is 3 and R' and R'' are both CH3, or R' is CH3 and R'' is hydroxylalkyl, or R' and R'' together with the N to which they are attached are a. a 5-membered heterocycle, the heterocycle being optionally substituted with OH; or b. Forming any six-membered heterocycle further containing one O heteroatom); R 4 is R 6 can be alkyl only when is -OH.

[0040] One embodiment of the present invention is R 4 is selected from the following: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. hydroxyl-substituted cycloalkylalkyl; v. 4-membered heterocycles 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 oxazoleheteroaryl; viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole. ix.-(CH2) n S(O)2CH3 or (CH2) n CN (wherein n is 3), x.-(CH2) n C(O)NR'R'' where n is 3 and R' and R'' are both CH3, or R' is CH3 and R'' is hydroxylalkyl, or R' and R'' together with the N to which they are attached are a. a 5-membered heterocycle, the heterocycle being optionally substituted with OH; or b. Forming any six-membered heterocycle further containing one O heteroatom); R 4 is R 6 can be alkyl only when is -OH.

[0041] One embodiment of the present invention is R 4 is selected from the following: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkyl substituted with hydroxy or alkoxy; v. Cycloalkylalkyl substituted with hydroxyl, alkoxy or -COOH; vi. 4- to 6-membered heterocycles containing a single O heteroatom; vii.(CH2) n C(O)OH, (CH2) n C(O)OCH3, or (CH2) n C(O)NHCH3 (wherein n is 2 or 3), viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazoleheteroaryl. R 4 is R 6 can be alkyl only when is -OH.

[0042] One embodiment of the present invention is R 4 is selected from the following: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. 4-membered heterocycles 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 oxazoleheteroaryl; R 4 is R 6 can be alkyl only when is -OH.

[0043] One embodiment of the present invention is R 6 is H.

[0044] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached, i. a 4- to 6-membered heterocycle containing a single O heteroatom, or ii. Forms any of 4- to 5-membered cycloalkyl rings; R 2 and R 3 is selected from H and alkyl, and R 2 or R 3 Only one of may be H or alkyl; R 4 is selected from: i. alkoxyalkyl, ii. cycloalkyl substituted with hydroxy or alkoxy; iii. cycloalkylalkyl substituted with hydroxy or alkoxy, and iv. 4-6 membered heterocycles containing a single O heteroatom; R 6 is H or -OH; Compounds and pharmaceutically acceptable salts are provided.

[0045] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached, i. a 4- to 6-membered heterocycle containing a single O heteroatom, or ii. Forms any of 4- to 5-membered cycloalkyl rings; R 2 and R 3 is selected from H and alkyl, and R 2 or R 3 Only one of may be H or alkyl; R 4 is selected from: i. cycloalkyl substituted with hydroxy or alkoxy; ii. cycloalkylalkyl substituted with hydroxy or alkoxy, and iii. 4- to 6-membered heterocycles containing a single O heteroatom; R 6 is H or -OH; Compounds and pharmaceutically acceptable salts are provided.

[0046] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached, i. a 4- to 6-membered heterocycle containing a single O heteroatom, or ii. Forms any of 4- to 5-membered cycloalkyl rings; R 2 and R 3 is selected from H and alkyl, and R 2 or R 3 Only one of may be H or alkyl; R 4 is selected from: i. cycloalkyl substituted with hydroxy or alkoxy, and ii. cycloalkylalkyl substituted with hydroxy or alkoxy; R 6 is H or -OH; Compounds and pharmaceutically acceptable salts are provided.

[0047] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached form a 4- to 6-membered heterocycle containing a single O heteroatom; R 2 and R 3 is selected from H and alkyl, and R 2 or R 3 Only one of may be H or alkyl; R 4 is selected from: i. hydroxy-substituted cycloalkyl, and ii. cycloalkylalkyl substituted with hydroxy; R 6 is H or -OH; Compounds and pharmaceutically acceptable salts are provided.

[0048] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached form a 4- to 6-membered heterocycle containing a single O heteroatom; R 2 is H and R 3 is alkyl; R 4 is selected from: i. hydroxy-substituted cycloalkyl, and ii. cycloalkylalkyl substituted with hydroxy; R 6 is H or -OH; Compounds and pharmaceutically acceptable salts are provided.

[0049] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached form a 4- to 6-membered heterocycle containing a single O heteroatom; R 2 is H and R 3 is alkyl; R 4 is selected from: i. hydroxy-substituted cycloalkyl, and ii. cycloalkylalkyl substituted with hydroxy; R 6 is H, Compounds and pharmaceutically acceptable salts are provided.

[0050] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached form a five-membered heterocycle containing a single O heteroatom; R 2 is H and R 3 is alkyl; R 4 is selected from: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. 4-membered heterocycles 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 oxazoleheteroaryl; R 6 is H or -OH, R 4 is R 6 may be alkyl only if -OH; Compounds and pharmaceutically acceptable salts are provided.

[0051] One embodiment of the present invention is a compound according to formula I, wherein: R 1 and R 5 , and the atoms to which they are attached form a five-membered heterocycle containing a single O heteroatom; R 2 is H and R 3 is methyl; R 4 is selected from: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. 4-membered heterocycles 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 oxazoleheteroaryl; viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; ix.-(CH2) n S(O)2CH3 or (CH2) n CN (wherein n is 3), x.-(CH2) n C(O)NR'R'' where n is 3 and R' and R'' are both CH3, or R' is CH3 and R'' is hydroxylalkyl, or R' and R'' together with the N to which they are attached are a. a 5-membered heterocycle, the heterocycle being optionally substituted with OH; or b. Forming any six-membered heterocycle further containing one O heteroatom); R 6 is H or -OH, R 4 is R 6 may be alkyl only if -OH; Compounds and pharmaceutically acceptable salts are provided.

[0052] A specific example of a compound of Formula I described herein is 5-[3-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol or a pharmaceutically acceptable salt thereof.

[0053] Other specific examples of compounds of formula I described herein include: 5-[5-methyl-3-[[rac-(3R)-1-(2-hydroxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formate salt; 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidyl]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-[[rac-(3R)-1-(2-methoxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidyl]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-piperidyl]butanoic acid; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]; Methyl 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoate; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N-methyl-butanamide; 5-[5-methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; and pharmaceutically acceptable salts thereof.

[0054] Other specific examples of compounds of formula I described herein include: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N,N-dimethyl-butanamide; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-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-piperidyl]-N-(2-hydroxyethyl)-N-methyl-butanamide; 1-(3-Hydroxypyrrolidin-1-yl)-4-[rac-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butan-1-one; Formic acid; 1-(3-hydroxypyrrolidin-1-yl)-4-[rac-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butan-1-one; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-1-morpholino-butan-1-one; 5-[5-methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidyl]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-piperidyl]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-piperidyl]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-piperidyl]butanenitrile; 5-[5-methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidyl]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-piperidyl]-N,N-dimethyl-butanamide; Formic acid'' 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N,N-dimethyl-butanamide; Formic acid; 5-[5-methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-methyl-3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; and pharmaceutically acceptable salts thereof.

[0055] A more preferred example of a compound of formula I described herein is 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoic acid or a pharmaceutically acceptable salt thereof.

[0056] Other more preferred examples of compounds of formula I described herein are: 5-[5-methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidyl]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-piperidyl]-N,N-dimethyl-butanamide; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-1-pyrrolidin-1-yl-butan-1-one; and pharmaceutically acceptable salts thereof.

[0057] Another embodiment of the present invention provides pharmaceutical compositions or medicaments containing a compound of the present invention and a therapeutically inert carrier, diluent, or excipient, as well as methods of using the compounds of the present invention to prepare such compositions and medicaments. In one example, a compound of Formula I can be formulated into a galenic dosage form by mixing it at ambient temperature, at an appropriate pH, and to the desired degree of purity with a physiologically acceptable carrier, i.e., a carrier that is nontoxic to recipients at the dosages and concentrations used. The pH of the formulation will depend primarily on the particular application and compound concentration, but is preferably in the range of about 3 to about 8. In one example, a 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.

[0058] The compositions are formulated, dosed, and administered in a manner consistent with good medical practice. Factors to consider in this regard include the particular disorder being treated, the particular 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 administration schedule, and other factors known to physicians.

[0059] The compounds of the present invention can be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural, and intranasal, and, if local treatment is desired, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0060] The compounds of the present invention can be administered in any convenient dosage form, such as, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions can contain conventional ingredients in pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and additional active agents.

[0061] Typical preparation is prepared by mixing the compound of the present invention with carrier or excipient.Suitable carrier and excipient are well known to those skilled in the art, and are described in detail in, for example, Ansel, Howard C., et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems.Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al., Remington: The Science and Practice of Pharmacy.Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients.Chicago, Pharmaceutical Press, 2005. The formulation may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricating agents, emulsifying agents, suspending agents, preservatives, antioxidants, opacifying agents, glidants, processing aids, colorants, sweeteners, flavors, flavorings, diluents, and other known additives to present the drug (i.e., a compound of the present invention or a pharmaceutical composition thereof) aesthetically or to aid in the manufacture of a pharmaceutical product (i.e., a medicament).

[0062] The compounds of formula I and their pharmaceutically acceptable salts can be processed with pharmaceutically inert, inorganic or organic adjuvants for the production of tablets, coated tablets, sugar-coated tablets, hard gelatin capsules, injection solutions or topical preparations.Lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc. can be used as such adjuvants for tablets, sugar-coated tablets and hard gelatin capsules.

[0063] Suitable adjuvants for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semisolid substances, and liquid polyols.

[0064] Suitable adjuvants for the production of solutions and syrups are, for example, water, polyols, saccharose, invert sugars, glucose etc.

[0065] Suitable adjuvants for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils and the like.

[0066] Suitable adjuvants for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solid or liquid polyols etc.

[0067] Suitable adjuvants for topical ophthalmic formulations are, for example, cyclodextrins, mannitol or many other carriers and excipients known in the art.

[0068] In addition, pharmaceutical preparations may contain preservatives, solubilizers, viscosity-increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavoring agents, salts for varying osmotic pressure, buffers, masking agents, or antioxidants. They may also contain still other therapeutically valuable substances.

[0069] The dosage may vary widely and, of course, be adapted to the individual requirements of each particular case. Generally, for oral administration, a daily dosage of about 0.1 mg to 20 mg per kg of body weight, preferably about 0.5 mg to 4 mg per kg of body weight (e.g., about 300 mg per person), is preferably divided into 1 to 3 individual doses, which may, if appropriate, consist of, for example, equal amounts. For topical administration, the formulation may contain 0.001% to 15% by weight of the drug, and the required dose, which may be between 0.1 and 25 mg, may be administered either as a single dose per day or per week, as multiple doses (2 to 4 times) per day, or as multiple doses per week. However, it will be apparent that the upper or lower limits given herein may be exceeded when indicated.

[0070] One embodiment of the present invention is a compound according to Formula I described herein for use as a therapeutically active substance.

[0071] One embodiment of the present invention is a compound according to Formula I as described herein for use in the treatment or prevention of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

[0072] One embodiment of the present invention is a compound according to Formula I as described herein for the treatment or prevention of a disease, disorder or condition, wherein the disorder or condition is responsive to NLRP3 inhibition.

[0073] As used herein, the term "NLRP3 inhibition" refers to a complete or partial reduction in the activity level of NLRP3, including, for example, inhibition of active NLRP3 and / or inhibition of NLRP3 activation.

[0074] There is evidence for a role for NLRP3-induced IL-1 and IL-18 in the inflammatory responses associated with or resulting from a number of different disorders (Menuet et al., Clinical and Experimental Immunology, 166:1-15, 2011; Strowiget et al., Nature, 481:278-286, 2012).

[0075] In one embodiment, the disease, disorder or condition is selected from the following: (i) Inflammation; (ii) autoimmune diseases; (iii) cancer; (iv) infectious diseases; (v) metabolic diseases; (vi) cardiovascular disease; (vii) respiratory diseases; (viii) liver disease; (ix) renal disease; (x) eye diseases; (xi) skin diseases; (xii) lymphatic status; (xiii) graft-versus-host disease; (xiv) allodynia; (xv) diabetes-related conditions; and (xvi) Any disease in which an individual is determined to have a germline or somatic non-silent mutation in NLRP3.

[0076] In another embodiment, the disease, disorder or condition is selected from the following: (i) Cancer; (ii) infectious diseases; (iv) cardiovascular disease; (v) liver disease; (vi) respiratory diseases; (vi) eye disease; and (vii) Skin diseases.

[0077] In another embodiment, the disease, disorder or condition is selected from the following: (i) gout; and (ii) Arthritis.

[0078] In a further exemplary embodiment of the invention, the disease, disorder or condition is inflammation. Examples of inflammation that may be treated or prevented include inflammatory responses associated with or resulting from: (i) skin conditions such as contact sensitivity, bullous pemphigoid, sunburn, psoriasis, atopic dermatitis, contact dermatitis, allergic contact dermatitis, seborrheic dermatitis, lichen planus, scleroderma, pemphigus, epidermolysis bullosa, urticaria, erythema, or alopecia; (ii) joint conditions such as osteoarthritis, systemic juvenile idiopathic arthritis, adult-onset Still's disease, relapsing polychondritis, rheumatoid arthritis, juvenile chronic arthritis, gout, or seronegative spondyloarthropathy (e.g., ankylosing spondylitis, psoriatic arthritis, or Reiter's disease); (iii) muscle conditions such as polymyositis or myasthenia gravis; (iv) gastrointestinal conditions 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 have effects remote from the gut (e.g., migraine, rhinitis, or eczema); (v) respiratory conditions such as chronic obstructive pulmonary disease (COPD), asthma (including eosinophilic, bronchial, allergic, intrinsic, extrinsic, or dust-induced asthma, especially chronic or refractory asthma, e.g., late-stage asthma and airway hyperresponsiveness), bronchitis, rhinitis (including acute rhinitis, allergic rhinitis, atrophic rhinitis, chronic rhinitis, caseous rhinitis, hypertrophic rhinitis, panlentic rhinitis, rhinitis sicca, rhinitis medicamentosa, membranous rhinitis, seasonal rhinitis, e.g., hay fever and vasomotor rhinitis), sinusitis, idiopathic pulmonary fibrosis (IPF), sarcoidosis, farmer's lung, silicosis, asbestosis, volcanic ash-induced inflammation, adult respiratory distress syndrome, hypersensitivity pneumonitis, or idiopathic interstitial pneumonia; (vi) vascular conditions such as atherosclerosis, Behçet's disease, vasculitis, or Wegener's granulomatosis; (vii) autoimmune conditions such as systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, Hashimoto's thyroiditis, type I diabetes, idiopathic thrombocytopenic purpura, or Graves' disease; (viii) ocular conditions such as uveitis, allergic conjunctivitis, or vernal conjunctivitis; (ix) an infectious disease or infection-related condition such as acquired immunodeficiency syndrome (AIDS), acute or chronic bacterial infection, acute or chronic parasitic infection, acute or chronic viral infection, acute or chronic fungal infection, meningitis, hepatitis (A, B, or C, or other viral hepatitis), peritonitis, pneumonia, epiglottitis, malaria, hemorrhagic dengue fever, leishmaniasis, streptococcal myositis, mycobacterium tuberculosis (including mycobacterium tuberculosis and HIV coinfection), mycobacterium avium intracellulare, pneumocystis carinii pneumonia, orchitis / epidydimitis, Legionnaires' disease, Lyme disease, influenza A, Epstein-Barr virus infection, viral encephalitis / aseptic meningitis, or pelvic inflammatory disease; (x) renal conditions such as mesangial proliferative glomerulonephritis, renal syndrome, nephritis, glomerulonephritis, obesity-related glomerulopathy, acute renal failure, acute kidney injury, uremia, nephritic syndrome, renal fibrosis including chronic crystalline nephropathy, or renal hypertension; (xi) lymphatic conditions such as Castleman's disease; (xii) a condition of the immune system, or a condition involving the immune system, such as hyper-IgE syndrome, lepromatous leprosy, familial hemophagocytic lymphohistiocytosis, or graft-versus-host disease; (xiii) liver conditions such as chronic active hepatitis, nonalcoholic steatohepatitis (NASH), alcohol-induced hepatitis, nonalcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease (AFLD), alcoholic steatohepatitis (ASH), primary biliary cirrhosis, fulminant hepatitis, hepatic fibrosis, or liver failure; (xiv) Cancer, including those mentioned above; (xv) burns, wounds, trauma, hemorrhage, or stroke; (xvi) radiation exposure; (xvii) metabolic diseases such as type 2 diabetes (T2D), atherosclerosis, obesity, gout, or pseudogout; and / or (xviii) Pain such as inflammatory hyperalgesia, pelvic pain, allodynia, neuropathic pain, or cancer-induced bone pain.

[0079] One embodiment of the present invention is a compound according to formula I described herein for the treatment or prevention of a disease, disorder or condition selected from the following: (i) Inflammation; (ii) autoimmune diseases; (iii) cancer; (iv) infectious diseases; (v) metabolic diseases; (vi) cardiovascular disease; (vii) respiratory diseases; (viii) liver disease; (ix) renal disease; (x) eye diseases; (xi) skin diseases; (xii) lymphatic status; (xiii) psychological disorders; (xiv) graft-versus-host disease; (xv) allodynia; (xvi) diabetes-related conditions; and (xvii) Any disease in which an individual is determined to have a germline or somatic non-silent mutation in NLRP3.

[0080] One embodiment of the invention is the use of a compound according to formula I as described herein in the treatment or prophylaxis of a disease, disorder or condition, which disease, disorder or condition is responsive to NLRP3 inhibition.

[0081] One embodiment of the present invention is the use of a compound according to formula I as described herein for use in the treatment or prevention of a disease, disorder or condition selected from asthma and COPD.

[0082] One embodiment of the present invention is the use of a compound according to formula I as described herein for use in the treatment or prevention of a cardiovascular disease, disorder or condition.

[0083] One embodiment of the present invention is the use of a compound according to formula I as described herein for use in the treatment or prevention of a disease, disorder or condition selected from cryopyrin-associated periodic syndromes.

[0084] One embodiment of the present invention is a compound according to Formula I as described herein for the treatment or prevention of a disease, disorder or condition selected from asthma and COPD.

[0085] One embodiment of the present invention is a compound of Formula I, as described herein, for treating or preventing a cardiovascular disease, disorder or condition.

[0086] One embodiment of the present invention is a compound according to Formula I as described herein for treating or preventing a disease, disorder or condition selected from cryopyrin-associated periodic syndromes.

[0087] One embodiment of the present invention is the use of a compound according to 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.

[0088] One embodiment of the present invention is the use of a compound according to formula I as described herein for the preparation of a medicament for treating or preventing a cardiovascular disease, disorder or condition.

[0089] One embodiment of the present invention is the use of a compound according to 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.

[0090] 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, comprising administering an effective amount of a compound according to formula I described herein.

[0091] One embodiment of the present invention is a method for treating or preventing a cardiovascular disease, disorder or condition, comprising administering an effective amount of a compound of formula I, as described herein.

[0092] One embodiment of the present invention is a method for treating or preventing a disease, disorder, or condition selected from cryopyrin-associated periodic syndromes, comprising administering an effective amount of a compound of formula I described herein.

[0093] One embodiment of the present invention relates to a method of inhibiting NLRP3, comprising administering an effective amount of a compound according to Formula I described herein.

[0094] Also, one embodiment of the present invention is a compound of formula I, as described herein, when prepared according to any one of the processes described.

[0095] 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.

[0096] Assay procedure NLRP3 and pyroptosis It is well established that activation of NLRP3 leads to cell pyroptosis, a hallmark that plays a key role in the development of clinical disease (Yan-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 & 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 block pyroptosis and the release of pro-inflammatory cytokines (e.g., IL-1β) from cells.

[0097] THP-1 cells: culture and preparation THP-1 cells (ATCC No. TIB-202) were grown in RPMI containing L-glutamine (Gibco No. 11835) supplemented with 1 mM sodium pyruvate (Sigma No. S8636) and penicillin (100 units / ml) / streptomycin (0.1 mg / ml) (Sigma No. P4333) in 10% fetal bovine serum (FBS) (Sigma No. F0804). Cells were passaged periodically until confluent (approximately 10 6The cells were grown to a concentration of 625,000 cells / ml. On the day of the experiment, THP-1 cells were harvested and resuspended in RPMI medium (without FBS). Cells were then counted, and viability (>90%) was confirmed using trypan blue (Sigma #T8154). Appropriate dilutions were made to obtain a concentration of 625,000 cells / ml. LPS (Sigma #L4524) was added to this diluted cell solution to obtain a final assay concentration (FAC) of 1 μg / ml. 40 μl of the final preparation was dispensed into each well of a 96-well plate. The plate thus prepared was used for compound screening.

[0098] THP-1 cell pyroptosis assay For compound screening, the following method stepwise assay was followed. Seed THP-1 cells (25,000 cells / well) in 40 μl of RPMI medium (without FBS) containing 1.0 μg / ml LPS in 96-well poly-D-lysine (VWR no. 734-0317) coated black-walled, clear-bottom cell culture plates. 5 μl of compound (8-point half-log dilutions with a top dose of 10 μM) or vehicle (DMSO 0.1% FAC) is added to the appropriate wells. Incubate at 37°C, 5% CO2 for 3 hours. Add 5 μl of nigericin (Sigma No. N7143) (FAC 5 μM) to all wells Incubate at 37°C, 5% CO2 for 1 hour. At the end of the incubation period, spin the plate at 300 x g for 3 minutes and remove the supernatant. Then, 50 μl of resazurin (Sigma #R7017) (100 μM resazurin in RPMI medium without FBS) is added and the plate is incubated at 37°C and 5% CO for a further 1-2 hours. Plates were read on an Envision reader at Ex 560nm and Em 590nm I C 50 Fit the data to a nonlinear regression equation (log inhibitor vs. response variable slope, 4 parameters) The results of the pyroptosis assay were compared with those of THP IC 50 These are summarized in Table 1 below.

[0099] Human whole blood IL-1β release assay For systemic delivery, the ability of a compound to inhibit NLRP3 when present in the bloodstream is crucial. Therefore, we investigated the NLRP3 inhibitory activity of a number of compounds in human whole blood according to the following protocol. Human whole blood in Li-heparin tubes was obtained from healthy donors from a volunteer donor pool. Plate out 80 μl of whole blood containing 1 μg / ml LPS into a 96-well flat-bottom cell culture plate (Corning #3585). 10 μl of compound (8-point half-log dilutions at the highest dose of 10 μM) or vehicle (DMSO 0.1% FAC) is added to the appropriate wells. Incubate at 37°C, 5% CO2 for 3 hours. Add 10 μl of nigericin (Sigma No. N7143) (10 μM FAC) to all wells Incubate at 37°C, 5% CO2 for 1 hour. At the end of the incubation period, spin the plate at 300 x g for 5 min to pellet the cells, remove 20 µl of the supernatant, and add it to a 96-well v-bottom plate for IL-1β analysis (Note: These plates containing supernatant can be stored at -80 °C for analysis at a later date). IL-1β was measured according to the manufacturer's protocol (Perkin Elmer-AlphaLisa IL-1 Kit AL220F-5000). I C 50 Fit the data to a nonlinear regression equation (log inhibitor vs. response variable slope, 4 parameters) Human whole blood assay results HWB IC 50 These are summarized in Table 1 below.

[0100] Microsomal stability: Incubations with 1 μM test compound in microsomes (0.5 mg / mL) and the cofactor NADPH are performed in 96-well plates at 37°C using a TECAN (Tecan Group Ltd, Switzerland) automated liquid handling system. After a 10-minute preincubation step between microsomes and test compound, the enzymatic reaction is initiated by the addition of the cofactor. Aliquots of the incubation are removed at 1, 3, 6, 9, 15, 25, 35, and 45 minutes and quenched with 1:3 (v / v) acetonitrile containing an internal standard. The samples are then cooled and centrifuged, after which the supernatants are analyzed by LC-MS / MS2.

[0101] Metabolic stability in hepatocytes: Assay Description: Biological materials. Obtain cryopreserved hepatocytes [mouse, rat, rabbit, monkey, human (mixed sex)]. Hepatocyte viability after reconstitution is at least 80% throughout the study. Obtain ready-to-use rat / human HepatoPac® cultures [long-term hepatocyte co-cultures; pooled (human male n=5, female n=5)], stromal mouse fibroblasts (negative control; pooled), culture plates, application medium, and maintenance medium. Metabolism by Suspension Hepatocytes. Initially, pooled cryopreserved hepatocytes were reconstituted with prewarmed William's E medium containing 10% FCS, 0.05 mg / mL streptomycin, 50 U / mL penicillin, and 0.4 mM L-glutamine, as well as 0.01 mg / mL gentamicin, 0.048 mg / mL hydrocortisone, and 0.004 mg / mL insulin to a final suspension density of 1 × 10 cells / mL. Incubations were performed automatically using a Liquid Handling System (Tecan) equipped with a CO2 incubator with an orbital shaker. After adding test compound (e.g., 1 μM) to the wells (1 × 10 cells / well), the 96-well hepatocyte suspension culture plate was incubated at 37°C in 5% CO2. Samples were quenched at the designated time points by adding acetonitrile (containing the internal standard) to the incubation wells for up to 2 hours. Incubations of test substances (e.g., 1 μM, 0.1% v / v DMSO) performed in metabolic suspension assays using HepatoPac® are performed in 96-well plates containing either cocultures of adherent hepatocytes with mouse fibroblast control cells or control cells alone (5% CO2 atmosphere, 37°C). The incubation medium for human HepatoPac® is identical to that used in suspension stem cells. At designated time points (2, 18, 26, 48, 72, and 96 hours), all wells are quenched with ice-cold acetonitrile containing an internal standard. The samples are then appropriately centrifuged and the supernatants are analyzed by LC-MS / MS. n=1 or 2 incubations are performed.

[0102] hERG screening assay During the small molecule drug development process, cardiac arrhythmias are one of the most frequent side effects leading to drug failure. Such failures are often related to the drug's ability to inhibit the human ether-a-go-go-related gene (hERG) cardiac potassium channel. Therefore, no or low inhibition of the hERG cardiac potassium channel is considered beneficial.

[0103] cell The CHO-crelox hERG cell line (ATCC reference number PTA-6812, female Chinese hamster cells) was generated and validated at Roche. Ready-to-use frozen instant CHO-hERG cells were cryopreserved at Evotec (Germany) and used directly in the experiments.

[0104] Experimental solution The extracellular solution contained (in mM): NaCl 150; KCl 4; CaCl 1; MgCl 1; HEPES 10; pH 7.2-7.4 with NaOH, osmolality 290-330 mOsm. The internal solution contained (in mM): KCl 10; KF 100; NaCl 10; HEPES 10; EGTA 20; pH 7.0-7.4 with KOH, osmolality 260-300 mOsm.

[0105] electrophysiology The effects of compounds on hERG K+-current parameters will be evaluated at two concentrations in at least four cells.

[0106] hERG studies are performed using the automated patch clamp system SynchroPatch® 384 (Nanion Technologies GmbH, Germany). K+ currents are measured using the patch voltage clamp technique in the whole-cell configuration at 35-37°C.

[0107] Cells were held at a resting voltage of -80 mV and stimulated with the voltage pattern shown in Figure 1 (outward K at 35–37 °C). + The pulse pattern used to elicit the current) activated hERG channels at a stimulation frequency of 0.1 Hz (6 bpm) to conduct outward IK hERG currents.

[0108] Data analysis The amplitude of the IKhERG was recorded at each drug concentration and compared to vehicle control values ​​(set at 100%) to define fractional blocks. The concentration-response data were fitted with the following relationship: [Table 1]

[0109] Concentration-response curves were fitted by nonlinear regression analysis using the EworkBook suite (ID Business Solutions Ltd, UK). Data were fitted using a four-parameter logistic model (fit = (A + (B / (1 + ((x / C)^D)))), where A = 0 and B = 100).

[0110] hERG assay results are compared with hERG IC 20 The results are summarized in Table 2 below.

[0111] Transcellular P-gp assay: A typical assay uses transfected LLC-PK1 cells (porcine kidney epithelial cells) overexpressing human P-gp or mouse P-gp cultured on 96-well semi-permeable filter membrane plates, where these cells form a polarized monolayer with tight junctions, which act as a barrier between the apical and basolateral compartments.

[0112] P-gp is expressed in the apical membrane of the monolayer. Cell monolayer adhesion and P-gp functional activity are confirmed by the addition of the cell-impermeable marker Lucifer Yellow and the reference P-gp substrate edoxaban, respectively. J. Pharmacol. Exp. Ther., 2021, 376, 322-329.

[0113] PAMPA: PAMPA (Parallel Artificial Membrane Permeability Assay) is a first-order selective permeability screen for drug candidates. The PAMPA assay uses artificial phospholipid membranes to mimic transcellular absorption conditions. This assay determines permeability values ​​that can be used for compound optimization and ranking purposes and as input parameters for in silico models predicting intestinal absorption.

[0114] The donor concentration is measured at t-start (baseline) and compared to the donor and acceptor concentrations after a certain time period (t-end), and the extent of compound crossing the membrane is calculated.

[0115] Bacterial Reverse Mutation Assay (AMES): Testing of compounds is performed as outlined in this guideline: Test No. 471: Bacterial Reverse Mutation Test | OECD Guidelines for the Testing of Chemicals, Section 4: Health Effects | OECD iLibrary (oecd-ilibrary.org)

[0116] Bacterial culture: The bacterial strains used are TA98, TA100, TA1535, TA97a, and TA102. A batch of each strain is maintained as a frozen stock. A vial is thawed and used to inoculate a culture in nutrient broth. The culture is placed in an incubator set at 37 °C with agitation for approximately 10 h to obtain a working culture of at least 10 cells / mL.

[0117] To ensure that the cultures are at the appropriate stage of growth and confluency, samples are taken from each culture at the end of the incubation period and assessed for confluency either by viability plating or OD650 assessment.

[0118] treatment: Triplicates are included for each concentration of compound and positive control and sextuplicates for vehicle control.

[0119] Formulations are prepared using DMSO to allow for the solubility limit or maximum exposure of freely soluble test substances up to 1000 μg / well, which is equivalent to the 5000 μg / plate used in the conventional plate-integrated Ames assay.

[0120] Concentrations are typically separated by half-log intervals in a single experiment: for soluble compounds, the concentrations are 0, 3.2, 10, 32, 100, 320, 1000 μg / well.

[0121] The positive controls used are as follows: Abbreviation Name Used for strain 2NF 2-Nitrofluorene TA98-S-9 NaN3 Sodium Azide TA100 and TA1535-S-9 AAC 9-aminoacridine TA97 aS-9 MMC Mitomycin C TA102-S-9 B[a]P Benzo[a]pyrene TA98+S-9 AAN Aminoanthracene TA100, TA1535, TA97a and TA102+S-9

[0122] Plating is achieved by the following sequential additions to 400 µL of supplemented molten agar at 45 ± 1 °C: 20 μL of bacterial culture 20 μL of test substance solution / vehicle control / positive control 100 μL of 10% S-9 mix or buffer solution It is then mixed quickly and poured into the mutagenesis plates (wells).

[0123] To set, invert the plate and incubate in an incubator set to 37 °C, protected from light, for 2-3 days.

[0124] toxicity: Toxicity is detected by the following parameters: · Reduction in background loans Significant reduction in revertants compared to concurrent vehicle controls · Decreased mutagenic response.

[0125] Scoring: Bacterial colonies are scored manually or electronically using an automated colony counter.

[0126] In vitro mammalian cell micronucleus assay: Testing of compounds is performed 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)

[0127] Cell culture: Cultures are maintained in tissue culture flasks containing HEPES-buffered RPMI 1640 medium containing GlutaMAX-1 supplemented with 10% (v / v) heat-inactivated fetal bovine serum, 100 units / mL penicillin / streptomycin, and 100 µg / mL penicillin / streptomycin in a humidified incubator set at 37 °C and 5% (v / v) CO in air. Cells are subcultured at low to medium density at least once before treatment.

[0128] The day before treatment, cells are subcultured at a density of approximately 7 x 104 cells / mL. Cells are maintained at 37°C, 5% (v / v) CO2 in air in a humidified environment prior to treatment.

[0129] treatment: Cultured human lymphoblastoid TK6 cells are exposed to compounds for 3 hours in the presence of S-9, followed by a 24-hour recovery period. Furthermore, a continuous 27-hour treatment in the absence of S-9 is included because some chemicals have been reported to only exert positive effects after prolonged treatment. This corresponds to approximately 1.5-2.0 times the average generation time of TK6 cells used in this laboratory (cell cycle time of approximately 15 hours). All cultures are sampled 27 hours after the start of treatment.

[0130] Prepare dilutions in DMSO that allow maximum exposure up to the solubility limit, 1 mM or 500 μg / mL, whichever is lower.

[0131] Typically, at least 12 concentrations are spaced 0.7-fold apart from the upper limit (for soluble compounds with MW ≥ 500, the concentrations are 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 DMSO concentration is 1% v / v. Positive controls are noscapine in the absence of S-9 and cyclophosphamide in the presence of S-9. Duplicate concentrations of compound and multiple concurrent vehicle and positive controls are included per treatment in a 96-well plate and incubated at 37°C, 5% (v / v) CO2 for the treatment period. The 3-h treatment cultures are washed once and re-incubated with fresh medium for 24 h.

[0132] Collection: At the specified sampling times, an aliquot of cell suspension from the designated culture will be taken for cell number determination 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 / glacial acetic acid (7:1 v / v). The fixed cells will be stored in fixative at 2–8°C before slide preparation.

[0133] Slides are air-dried before staining, immersed in 12.5 μg / mL acridine orange in phosphate-buffered saline (PBS), pH 6.8, for approximately 10 minutes, followed by a few seconds of rinsing (with agitation) in PBS.

[0134] Cytotoxicity readout and concentration selection: Toxicity is expressed as population doubling (PD) compared to vehicle control. PD is calculated for each concentration as follows: PD=[log(N / X0)] / log2 where N = average final cell number / culture at each concentration X0 = starting (baseline) count

[0135] The highest concentration for micronucleus analysis should not exceed (approximately) 50% cytotoxicity, be the highest concentration tested, or be the lowest precipitating concentration observed by eye at the end of the treatment incubation period.

[0136] Analyze slides from the highest selected concentration and at least two lower concentrations, covering the range from maximum to little or no cytotoxicity, as needed. A minimum of 1000 mononuclear cells from each culture (2000 per concentration) will be analyzed for micronuclei.

[0137] Evaluation criteria: A compound is considered to induce chromosomal abnormalities and / or aneuploidy events if: - A statistically significant increase in the frequency of MNMON cells at one or more concentrations is observed. - The incidence of cells with such a concentration of micronuclei exceeds the normal range in both replicates. A concentration-related increase in the percentage of cells with micronuclei is observed (positive trend test).

[0138] A compound is considered positive in this assay if all of the above criteria are met.

[0139] If none of the above criteria are met, the compound is considered negative in this assay.

[0140] Results that only partially meet the above criteria will be treated on a case-by-case basis, but in the context of a screening study, will be concluded as either positive, negative, or equivocal. Evidence of a concentration-related effect is considered useful but not essential for the evaluation of a positive result. Biological relevance takes into account, for example, consistency of response within and between concentrations and between experiments (if applicable), or effects that occur only at highly toxic concentrations.

[0141] Pharmacokinetic profile of test article in minipigs: The pharmacokinetics of the test substance was determined in minipigs after intravenous and oral administration. The experimental design consisted of three male minipigs, each of which received a single intravenous bolus dose and a single oral dose with the test substance. The intravenous dose was administered at a nominal dose volume of 1 mL / kg. The oral dose was administered by gastric gavage at a nominal dose volume of 5 mL / kg. There was a washout period of at least 7 days between the last sampling and the next dose in the same animal. The content of all formulations was within the desired range of 85-115% of the nominal content. After administration, blood samples (1 mL) were collected from the saphenous or jugular vein of each animal pre-dose, 5, 15, and 30 minutes after IV administration, 1, 2, 4, 8, and 24 hours after administration, and 15, 30 minutes, 1, 2, 4, 6, 8, 24, and 48 hours after oral administration. Hematocrit was determined at all time points. Blood:plasma partition coefficients were determined at 2 and 4 hours, and urine was collected as a single sample over 24 hours after dose administration. Blood samples (nominal 1 mL) were collected from each animal's saphenous vein (via cannula) or jugular vein into polypropylene tubes containing K2EDTA anticoagulant and centrifuged (1500 g, 10 minutes, 4°C) to prepare plasma for analysis. Residual blood cells were discarded. Plasma vials were capped and stored on wet ice for ≤60 minutes before being transferred to <-50°C storage (nominal -80°C) before analysis by specific LC-MS methods.

[0142] Toxicity evaluation of test substances in minipigs The maximum tolerated dose (MTD) of the test article is determined after a single daily oral (gavage) administration to minipigs. The toxicity of repeated daily administration for 14 days is then assessed. Furthermore, the toxicokinetic profile of the test article is characterized. Purpose-bred Gottingen minipigs are obtained from Ellegaard Gottingen, Dalmoos, Denmark (animals: age range: 2-3 months, body weight range: 4-6 kg). At the start of dosing, animals are 4-5 months old and weigh between 6-9.5 kg. A dose volume of 10 mL / kg is used. Individual dose volumes are calculated from each animal's most recent body weight up to 30, 100, and 300 mg / kg / day or other target dose levels depending on non-MTD results. Blood samples are collected on days 1 and 14 to determine plasma drug concentrations and derived toxicokinetic parameters. Animals are not fed on the day of the scheduled necropsy. Each animal was anesthetized by intramuscular injection of the Zoletil mixture and then sacrificed by exsanguination. All tissues were preserved in appropriate fixatives. Further analysis included food consumption, body weight, clinical pathology, and complete histopathological examination of target organs. [Table 2] [Table 3] [Table 4]

[0143] The invention will now be illustrated by the following examples, which have no limiting character.

[0144] Where preparations are obtained as mixtures of enantiomers or diastereomers, the pure enantiomers or diastereomers may be obtained by methods described herein or by methods known to those skilled in the art, such as, for example, chiral chromatography or crystallization.

[0145] Experimental Method [Table 5] [Example]

[0146] Unless otherwise specified, all examples and intermediates were prepared under a nitrogen atmosphere.

[0147] Example 1: 5-[3-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0148] Step A: 4-benzyloxy-5-bromo-2,3-dihydrobenzofuran To a solution of 5-bromocoumaran-4-ol (CAS no. 2279149-27-6, 4.59 g, 20.26 mmol, 1.00 equiv.) in acetonitrile (40 mL) was added potassium carbonate (5.6 g, 40.51 mmol, 2.00 equiv.), followed by benzyl bromide (4.89 g, 3.4 mL, 28.57 mmol, 1.41 equiv.). The reaction mixture was stirred at room temperature for 2 hours. 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 (6.17 g, 95% yield) as a colorless oil. LCMS: m / z 305.1 / 307.0 [M+H] + ,ESI pos.

[0149] Step B: 2-(4-benzyloxy-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane [ka] To a solution of 4-benzyloxy-5-bromo-2,3-dihydrobenzofuran (Example 1, Step A) (6.16 g, 19.18 mmol, 1.00 equiv.) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS No. 61676-62-8, 5.47 g, 6.0 mL, 29.41 mmol, 1.53 equiv.) in tetrahydrofuran (80 mL) was added n-butyllithium, a 1.6 M solution in hexanes (19 mL, 30.4 mmol, 1.59 equiv.), dropwise at −76° C. within 40 minutes. The mixture was stirred at −76° C. for 2.5 hours. The reaction mixture was warmed to −60° C., quenched with saturated aqueous NH4Cl at −60° C., 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 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, 120 g, gradient 0% to 10% ethyl acetate in heptane) to afford the title compound (5.78 g, 81% yield) as a colorless oil. LCMS: m / z 353.1 [M+H] + ,ESI pos.

[0150] Step C: 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol [ka] A solution of 2-(4-benzoxycoumaran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (Example 1, Step B) (5.77 g, 15.56 mmol, 1.00 equiv.) in ethyl acetate (70 mL) was alternately evacuated and flushed with argon three times. Palladium on activated carbon, 10% Pd based (577 mg, 0.54 mmol, 0.03 equiv.) was added. The reaction flask was evacuated, flushed with argon, evacuated, and flushed with hydrogen. The reaction mixture was stirred under a hydrogen atmosphere (balloon) at room temperature for 3 hours. Methanol (10 mL) was added. The reaction flask was alternately evacuated, flushed with argon, evacuated, and then flushed with hydrogen three times. The reaction mixture was stirred under a hydrogen atmosphere (balloon) at room temperature for 1 hour. The reaction mixture was filtered and rinsed thoroughly with ethyl acetate / methanol. The filtrate was concentrated in vacuo to give the title compound (4.22 g, 98% yield) as an off-white solid, which was used without further purification. LCMS: m / z 263.2 [M+H] + ,ESI pos.

[0151] Step D: tert-butyl (3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate [ka] To a mixture of 3,6-dichloro-5-methyl-1,2,4-triazine (CAS No. 132434-82-3, 180 mg, 1.10 mmol, 1.00 equiv.) and tert-butyl (3R)-3-aminopiperidine-1-carboxylate (CAS No. 188111-79-7, 330 mg, 1.65 mmol, 1.50 equiv.) in 1,4-dioxane (3.6 mL) was added N,N-diisopropylethylamine (148 mg, 0.20 mL, 1.15 mmol, 1.04 equiv.). The reaction mixture was stirred at room temperature for 16 hours. 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 0% to 40% ethyl acetate in heptane). All product-containing fractions were combined and concentrated in vacuo to give the title compound (351 mg, 93% yield) as a yellow oil. LCMS: m / z 328.3 [M+H] + ,ESI pos.

[0152] Step E: 6-chloro-5-methyl-N-[(3R)-3-piperidyl]-1,2,4-triazin-3-amine [ka] 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 equiv.) 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 equiv.). The reaction mixture was stirred at room temperature for 1 hour. 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 give the title compound (95 mg, 91% yield) as an orange foam. LCMS: m / z 228.1 [M+H] + ,ESI pos.

[0153] Step F: 2-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]ethanol [ka] To a solution of 6-chloro-5-methyl-N-[(3R)-3-piperidyl]-1,2,4-triazin-3-amine (Example 1, Step E) (95 mg, 0.40 mmol, 1.00 equiv.) in tetrahydrofuran (1.6 mL) was added 2-iodoethanol (CAS No. 624-76-0, 84 mg, 0.038 mL, 0.49 mmol, 1.23 equiv.), followed by N,N-diisopropylethylamine (133 mg, 0.180 mL, 1.03 mmol, 2.60 equiv.). The reaction mixture was stirred at 40° C. for 16 hours. 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 give the title compound (61 mg, 54% yield) as a pale yellow oil. LCMS: m / z 272.2 [M+H] + ,ESI pos.

[0154] Step G: 5-[3-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka] A mixture of 2-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]ethanol (Example 1, Step F) (56 mg, 0.20 mmol, 1.00 equiv), 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 equiv), cesium carbonate (192 mg, 0.59 mmol, 3.01 equiv) and XPhos Pd G3 (25 mg, 0.03 mmol, 0.15 equiv) in 1,4-dioxane (1.2 mL) and water (0.30 mL) was flushed with argon and stirred at 100° C. for 2.5 hours. The reaction mixture was cooled to room temperature and extracted with ethyl acetate and half-saturated aqueous NH4Cl. 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, 0% to 100% gradient in dichloromethane (dichloromethane:methanol:NH4OH 9:1:0.05)) to afford the title compound (49 mg, 64% yield) as a yellow foam. LCMS: m / z 372.3 [M+H] + ,ESI pos.

[0155] Example 2: 5-[5-methyl-3-[[(3R)-1-(2-hydroxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0156] Step A: Methyl 2-benzyloxypropanoate To a solution of methyl 2-hydroxypropanoate (CAS: 547-64-8; 5.0 g, 48.03 mmol, 1.0 equiv.) in THF (90 mL) was added NaH (2882 mg, 60% in mineral oil, 72.05 mmol, 1.5 equiv.) and stirred at 0 °C for 10 min. Benzyl bromide (CAS: 100-39-0; 5.71 mL, 48.0 mmol, 1.0 equiv.) was then added, and the reaction mixture was warmed to room temperature and stirred for 1 h. The mixture was quenched with ice-water NH4Cl (150 mL) and extracted with EtOAc (100 mL × 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 (6.9 g, 74% yield) as a colorless oil. 1 H NMR(400MHz,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).

[0157] Step B: 2-(benzyloxy)propanal (CAS: 53346-05-7) To a solution of methyl 2-benzyloxypropanoate (1.0 g, 5.15 mmol, 1.0 equiv.) in DCM (40 mL), DIBAl-H (7.72 mL, 7.72 mmol, 1.5 equiv.) was added dropwise over 10 min at 78 °C under N2, and the reaction mixture was stirred at -78 °C for 1 h. Upon completion of the reaction, the mixture was quenched with ice-NH4Cl (100 mL) and extracted with DCM (100 mL x 2). The organic phase was washed with brine (100 mL x 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 afford the title compound (320.0 mg, 38% yield) as a colorless oil. 1 H NMR(400MHz,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).

[0158] Step C: Benzyl N-[(3R)-1-(2-benzyloxypropyl)-3-piperidyl]carbamate To a mixture of benzyl N-[(3R)-3-piperidyl]carbamate (CAS: 478646-32-1; 228 mg, 0.97 mmol, 0.5 equiv.) and acetic acid (117 mg, 1.95 mmol, 1.0 equiv.) in DCE (4 mL) / methanol (2 mL), 2-benzyloxypropanal (CAS: 53346-05-7; 320.0 mg, 1.95 mmol, 1.0 equiv.) was added and stirred for 10 min. NaBH(OAc) (2065 mg, 9.74 mmol, 5.0 equiv.) was then added and stirred at 25 °C under N for 2 h. Upon completion of the reaction, the reaction mixture was concentrated in vacuo. The residue was purified by column chromatography (SiO, EA / MeOH: 4 / 1) to afford the title compound (210 mg, 28% yield) as a colorless oil. LCMS: m / s 383.4 [M+H]+, ESI pos.

[0159] Step D: (3R)-1-(2-(benzyloxy)propyl)piperidin-3-amine To a solution of benzyl N-[(3R)-1-(2-benzyloxypropyl)-3-piperidyl]carbamate (210.0 mg, 0.55 mmol, 1.0 equiv) in methanol (4 mL) was added Pd / C (10 mg, 10% palladium on carbon), and the mixture was stirred under H (1100 mmHg) at 25 °C for 2 h. The suspension was filtered through a Celite pad, the pad was washed with MeOH (3 mL × 2), and the combined filtrate was concentrated in vacuo to give the title compound (136.0 mg, quantitative) as a yellow oil. LCMS: 249.2 [M+H], ESI, pos.

[0160] Step E: 1-[(3R)-3-amino-1-piperidyl]propan-2-ol A solution of (3R)-1-(2-(benzyloxy)propyl)piperidin-3-amine (136.0 mg, 0.55 mmol, 1.0 equiv), 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 70 °C for 16 hours under 4500 mmHg. The suspension was filtered through a Celite pad, and the pad was washed with MeOH (5 mL × 2). The combined filtrate was concentrated in vacuo to give the title compound (86 mg, 84% yield) as a yellow oil. 1H NMR(400MHz,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).

[0161] Step F: 1-((R)-3-((6-chloro-5-methyl-1,2,4-triazin-3-yl)amino)piperidin-1-yl)propan-2-ol To a solution of 1-[(3R)-3-amino-1-piperidyl]propan-2-ol (86.0 mg, 0.54 mmol, 1.0 equiv.) and 3,6-dichloro-5-methyl-1,2,4-triazine (CAS No. 132434-82-3, 89.13 mg, 0.54 mmol, 1.0 equiv.) in 1,4-dioxane (2 mL) was added DIEA (0.16 mL, 0.98 mmol, 1.8 equiv.). The reaction mixture was stirred at 25 °C for 16 h. Upon completion, the reaction mixture was filtered and concentrated in vacuo. The residue was purified by C18 column chromatography (water / 0.1% NH₃·H₂O in MeCN, MeCN: 54%-46%). The eluent was lyophilized to afford the title compound (27.0 mg, 17% yield) as a yellow oil. LCMS: m / z 286.1 [M+H] + ,ESI pos.

[0162] Step G: 5-[5-methyl-3-[[(3R)-1-(2-hydroxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol 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 equiv), 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 equiv), and CsF (10.98 mg, 0.19 mmol, 2.0 equiv) in 1 mL of 1,4-dioxane / 0.2 mL of water was added XPhos Pd G3 (16.01 mg, 0.02 mmol, 0.2 equiv) at 25° C., followed by stirring at 95° C. under N2 for 2 h. After the reaction was completed, the reaction mixture was cooled to room temperature. The combined filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (neutral) to give the title compound (8.31 mg, 21% yield) as a yellow solid. 1 H NMR(400MHz,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).

[0163] Example 3: 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0164] Step A: Benzyl N-[(3R)-1-(3-benzyloxypropyl)-3-piperidyl]carbamate To a solution of benzyl N-[(3R)-3-piperidyl]carbamate (CAS: 478646-32-1; 0.5 g, 2.13 mmol, 1.0 equiv.) and CHCOOH (12.8 mg, 0.21 mmol, 0.1 equiv.) in DCE (5 mL), 3-benzyloxypropanal (CAS: 19790-60-4; 420.5 mg, 2.56 mmol, 1.2 equiv.) was added and stirred at 20 °C for 5 min. NaBH(OAc) (904.6 mg, 4.27 mmol, 5.0 equiv.) was then added and 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 NaSO, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with (ethyl acetate / methanol=40:1 to 5:1) to give the title compound (700 mg, 84% yield) as a colorless oil. LCMS: m / z 383.1 [M+H] + ,ESI pos.

[0165] Step B: (3R)-1-(3-benzyloxypropyl)piperidin-3-amine To a solution of benzyl N-[(3R)-1-(3-benzyloxypropyl)-3-piperidyl]carbamate (700.0 mg, 1.83 mmol, 1.0 equiv.) in methanol (2 mL) was added Pd / C (10 mg, 10% palladium on carbon) and Pd(OH)2 / C (10 mg, 10% palladium on carbon) under N2. After purging with H2 three times, stirring was continued under 1100 mmHg of H2 at 40 °C for 2 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give the title compound (450 mg, 99% yield) as a colorless oil. LCMS: m / z 249.0 [M+H] + ,ESI pos.

[0166] Step C: 3-[(3R)-3-amino-1-piperidyl]propan-1-ol (CAS: 1704948-85-5) To a solution of (3R)-1-(3-benzyloxypropyl)piperidin-3-amine (450.0 mg, 2.01 mmol, 1.0 equiv) in methanol (10 mL) was added Pd / C (50 mg, 10% palladium on carbon) under N. After purging with H three times, stirring was continued under 4500 mmHg of H at 70 °C for 2 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give the title compound (318 mg, 92% yield) as a yellow oil.

[0167] Step D: 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]propan-1-ol To a solution of 3-[(3R)-3-amino-1-piperidyl]propan-1-ol (57.9 mg, 0.37 mmol, 1.2 equiv.), DIEA (0.08 mL, 0.46 mmol, 1.5 equiv.) 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 equiv.). The reaction mixture was stirred at 20° C. for 16 hours. The reaction mixture was concentrated under reduced pressure to give the crude product, which was purified by reverse-phase flash (CombiFlash 0.1% NH3HO aqueous-ACN condition) followed by lyophilization to give the title compound (20.0 mg, 22% yield) as a yellow solid. LCMS: m / z 286.2 [M+H] + ,ESI pos.

[0168] Step E: 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol 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 equiv.) and 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]propan-1-ol (10.0 mg, 0.03 mmol, 1.0 equiv.) in 1,4-dioxane (0.5 mL) / water (0.1 mL), CsF (4.07 mg, 0.07 mmol, 2.0 equiv.) was added, followed by XPhos Pd G3 (1.48 mg, 0.0 mmol, 0.05 equiv.) 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 flash (CombiFlash 0.1% NH3HCO3 water-ACN condition) to give the title compound (9.1 mg, 66% yield) as a yellow solid. LCMS: m / z 386.3 [M+H] + ,ESI pos.

[0169] Example 4: 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0170] Step A: Methyl 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carboxylate To a solution of methyl 1-hydroxycyclopropane-1-carboxylate (5.00 g, 43.06 mmol, 1.0 equiv; 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 equiv) and stirred at 0 °C for 20 min. 2-(Trimethylsilyl)ethoxymethyl chloride (8.38 mL, 47.36 mmol, 1.1 equiv) was added slowly and stirred at 25 °C for 1 h. The reaction mixture was diluted with NHCl (500 mL) and extracted with ethyl acetate (3 × 200 mL). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was then purified by column chromatography on silica gel (petroleum ether / ethyl acetate: 1 / 0 to 10 / 1) to give the title compound (2.10 g, 20% yield) as a yellow oil. 1 H NMR(400MHz,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).

[0171] Step B: (1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropyl)methanol To a solution of methyl 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carboxylate (1.10 g, 4.46 mmol, 1.0 equiv.) in DCM (20 mL) was added DIBAl-H (13.39 mL, 13.39 mmol, 3.0 equiv.) dropwise under N at −78 °C, and the mixture was stirred at −78 °C for 1 h. The reaction mixture was diluted with saturated NH Cl (100 mL) and DCM (20 mL) at 0 °C, filtered, and the filtrate was extracted with DCM (20 mL × 3). The combined organic phase was 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 (0.55 g, 2.5 mmol, 56% yield) as a yellow oil. 1H NMR(400MHz,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).

[0172] Step C: 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carbaldehyde To a solution of DMSO (0.23 mL, 3.3 mmol, 2.0 equiv) in DCM (7 mL) was added oxalyl chloride (0.28 mL, 3.30 mmol, 2.0 equiv) at −70° C. and 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 equiv) in DCM (2 mL) was added dropwise to the above solution and stirred at −70° C. for 0.5 h. TEA (1.8 mL, 12.93 mmol, 7.84 equiv) was added and stirring was continued at 0° C. for 30 min. The reaction mixture was warmed to room temperature. It was diluted with water (50 mL) and extracted with DCM (30 mL × 3). The combined organic phase was 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 column chromatography on silica gel (petroleum ether / ethyl acetate: 1 / 0 to 10 / 1) to give the title compound (0.18 g, 50% yield) as a yellow oil. 1 H NMR(400MHz,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).

[0173] Step E: tert-butyl (R)-3-((6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl)amino)piperidine-1-carboxylate To a solution of tert-butyl (3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylate (700 mg, 2.14 mmol, 1.0 equiv; prepared as described in Example 1, Step D), 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 equiv), and CsF (1.30 g, 8.54 mmol, 4.0 equiv) in 1,4-dioxane (20 mL) and water (4 mL) was added XPhos Pd G3 (0.362 g, 0.43 mmol, 0.2 equiv), and stirring was continued at 95° C. under a N2 atmosphere for 3 h. The reaction mixture was cooled to room temperature. The mixture was 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 give the title compound (0.750 g, 82% yield) as a yellow solid. 1 H NMR(400MHz,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).

[0174] Step F: (R)-5-(5-methyl-3-(piperidin-3-ylamino)-1,2,4-triazin-6-yl)-2,3-dihydrobenzofuran-4-ol 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 equiv) in DCM (3 mL) / TFA (3.0 mL) was stirred at 25 °C for 1 h. The solvent was removed by a stream of nitrogen to give the crude product, after which the pH was adjusted to approximately 7 with NH3·H2O solution. The crude product was purified by reverse-phase flash (0.1% NH3·H2O, water-MeCN) to give the title compound (0.346 g, 77% yield) as a yellow solid. LCMS: m / z 328.2 [M+H] + ,ESI pos.

[0175] 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 A mixture of 1-((2-(trimethylsilyl)ethoxy)methoxy)cyclopropane-1-carbaldehyde (0.171 g, 0.79 mmol, 1.3 equiv), AcOH (3.7 mg, 0.06 mmol, 0.1 equiv), (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 equiv) in DCE (2 mL) was stirred at 25 °C for 10 min, then NaBHCN (7.67 mg, 1.22 mmol, 2.0 equiv) was added and stirred at 40 °C for 1 h. The solvent was removed by a stream of nitrogen. The crude product was purified by reverse phase flash (0.1% NH3·H2O, water-ACN) to give the title compound (120 mg, 0.23 mmol, 37% yield) as a yellow solid. LCMS: m / z 528.4 [M+H] + ,ESI pos.

[0176] Step H: 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol 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 equiv) 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. The pH was then adjusted to 7 with NH3·H2O, and the crude product was purified by reverse-phase flash (0.1% NH3·H2O in water-ACN) to give the title compound (9.89 mg, 0.02 mmol, 31% yield) as a yellow solid. LCMS: m / z 398.3 [M+H] + ,ESI pos.

[0177] Example 5: 5-[3-[[(3R)-1-(2-Methoxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; Formate [ka]

[0178] Step A: Benzyl N-[(3R)-1-(2-methoxyethyl)-3-piperidyl]carbamate To a solution of benzyl N-[(3R)-3-piperidyl]carbamate (CAS No. 478646-32-1, 0.5 g, 2.13 mmol, 1.0 equiv.), N-ethyl-N-isopropylpropan-2-amine (0.75 mL, 4.27 mmol, 2.0 equiv.) in acetonitrile (5 mL), 2-bromoethyl methyl ether (CAS: 6482-24-2; 0.3 mL, 3.2 mmol, 1.5 equiv.) was added, and the reaction mixture was stirred at 50 °C for 6 hours. Upon completion of the reaction, 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 give the title compound (520.0 mg, 83% yield) as an orange solid. LCMS: m / z 293.1 [M+H] + ,ESI pos.

[0179] Step B: (3R)-1-(2-methoxyethyl)piperidin-3-amine To a solution of benzyl N-[(3R)-1-(2-methoxyethyl)-3-piperidyl]carbamate (470.0 mg, 1.61 mmol, 1.0 equiv) in methanol (5 mL) was added Pd / C (50.0 mg, 10% palladium on carbon) and Pd(OH) / C (52.22 mg, 10% palladium on carbon) under N. After purging with H three times, the mixture was then stirred under 1100 mmHg of H at 20 °C for 2 h. Upon completion of the reaction, the reaction mixture was filtered, and the mother liquor was concentrated in vacuo to afford the title compound (240.0 mg, 94% yield) as a yellow oil. 1H-NMR(400MHz,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).

[0180] Step C: 6-chloro-N-[(3R)-1-(2-methoxyethyl)-3-piperidyl]-5-methyl-1,2,4-triazin-3-amine To a solution of (3R)-1-(2-methoxyethyl)piperidin-3-amine (231.6 mg, 1.46 mmol, 1.2 equiv.), N-ethyl-N-isopropylpropan-2-amine (0.38 mL, 2.2 mmol, 1.8 equiv.) in 1,4-dioxane (3 mL), 3,6-dichloro-5-methyl-1,2,4-triazine (CAS#132434-82-3, 200.0 mg, 1.22 mmol, 1.0 equiv.) was added, and the reaction mixture was stirred at 20 °C for 16 hours. Upon completion of the reaction, 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 (30.0 mg, 9% yield) as a colorless oil. LCMS: m / z 286.2 [M+H]+, ESI pos.

[0181] Step D: 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; Formate salt To a solution of 6-chloro-N-[(3R)-1-(2-methoxyethyl)-3-piperidyl]-5-methyl-1,2,4-triazin-3-amine (30.0 mg, 0.1 mmol, 1.0 equiv.), 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 equiv.), and CsF (63.8 mg, 0.42 mmol, 4.0 equiv.) in 1,4-dioxane (1 mL) and water (0.2 mL) was added XPhos Pd G3 (17.8 mg, 0.02 mmol, 0.2 equiv.) under N2 at 20 °C. The reaction mixture was stirred at 80 °C for 2 hours. Upon completion of the reaction, the mixture was cooled to room temperature. The residue was filtered and concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex luna C 18 Purification by 150*25mm*10um; 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) gave the title compound (18.2 mg, 40% yield) as a brown gum. LCMS: m / z 386.1 [M+H] + ,ESI pos.

[0182] Example 6: (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 [ka]

[0183] Step A: tert-Butyl (3R,5S)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-5-hydroxy-piperidine-1-carboxylate To a solution of 3,6-dichloro-5-methyl-1,2,4-triazine (CAS No. 132434-82-3, 243 mg, 1.48 mmol, 1.0 equiv.) in 1,4-dioxane (5 mL) at ambient temperature was added (3R,5S)-3-amino-5-hydroxypiperidine-1-carboxylic acid tert-butyl ester (CAS No. 1932513-59-1, 396 mg, 1.78 mmol, 1.2 equiv.) and N,N-diisopropylethylamine (345 mg, 0.47 mL, 2.67 mmol, 1.8 equiv.) to give a dark brown solution. The reaction mixture was stirred at 23 °C for 60 h. The reaction mixture was then quenched with half-saturated aqueous NH4Cl (50 mL) and extracted with ethyl acetate (2 × 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, 0-80% ethyl acetate in heptane) to give the title compound (345 mg, 68% yield) as a pale yellow oil. LCMS: m / z 344.2 ([{35Cl}M+H] + ), 346.2([{37Cl}M+H] + ),ESI pos.

[0184] Step B: (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidin-3-ol; Hydrochloride To a solution of tert-butyl (3R,5S)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-5-hydroxy-piperidine-1-carboxylate (Example 6, Step A) (345 mg, 1.0 mmol, 1.0 equiv.) in dichloromethane (10 mL) and methanol (5 mL) was added 4 M HCl in 1,4-dioxane (3.01 g, 2.51 mL, 10.03 mmol, 10.0 equiv.) dropwise at room temperature. The reaction mixture was stirred at 23 °C for 16 hours. The reaction mixture was concentrated in vacuo to afford the title compound (312 mg, 100% yield) as a light brown foam, which was used without further purification. LCMS: m / z 244.1 ([{35Cl}M+H]+), 246.1 ([{37Cl}M+H]+), ESI pos.

[0185] Step C: (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-ethyl-piperidin-3-ol To a suspension of (3S,5R)-5-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidin-3-ol (Example 6, Step B) (312 mg, 1.0 mmol, 1.0 equiv.) in dichloromethane (15 mL) was added acetaldehyde (CAS No. 75-07-0; 110 mg, 0.14 mL, 2.51 mmol, 2.5 equiv.) at 0 ° C., followed by sodium acetate (CAS No. 127-09-3; 206 mg, 2.51 mmol, 2.5 equiv.) and sodium triacetoxyborohydride (CAS No. 56553-60-7; 382 mg, 1.8 mmol, 1.8 equiv.). 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 purified by flash chromatography (silica gel, 25 g, 0-100% gradient in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (137 mg, 50% yield) as a light brown foam. LCMS: m / z 272.1 ([{35Cl}M+H] + ), 274.1([{37Cl}M+H] + ),ESI pos.

[0186] 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 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 equiv) and 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 equiv) and potassium carbonate (205 mg, 1.48 mmol, 4.8 equiv) 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-palladium(II) dichloride dichloromethane complex (CAS number 95464-05-4, 38 mg, 0.046 mmol, 0.15 equiv.) was added. After flushing with argon again, the sealed tube was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature, quenched with water (50 mL) and saturated NH4Cl solution (50 mL), and 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 adsorbed 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 preparative HPLC (column: Gemini NX, 12 nm, 5 μm, 100 × 30 mm; MeCN / water (+0.1% TEA)) to afford the title compound (43 mg, 37% yield) as a yellow amorphous lyophilized solid. LCMS: m / z 372.2 [M+H] + ,ESI pos.

[0187] Example 7: 5-[5-methyl-3-[[(3R)-1-(2-methoxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0188] Step A: 2-Methoxypropyl ethanesulfonate To a solution of 2-methoxypropanol (1.10 g, 12.2 mmol, 1.0 equiv.) and DIEA (4.72 g, 36.6 mmol, 3.0 equiv.) in DCM (10 mL) was added ethanesulfonyl chloride (1.73 mL, 18.3 mmol) at 0° C. and then stirred at 20° C. for 1 h. Upon completion of the reaction, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phase was 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 afford the title compound (2.3 g, 98% yield) as a black solid. 1 H NMR(400MHz,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).

[0189] Step B: Benzyl N-[(3R)-1-(2-methoxypropyl)-3-piperidyl]carbamate To a solution of benzyl N-[(3R)-3-piperidyl]carbamate (CAS No. 478646-32-1, 500.0 mg, 2.13 mmol, 1.0 equiv.) and TEA (0.5 g, 4.91 mmol, 2.3 equiv.) in ACN (5 mL) was added 2-methoxypropyl ethanesulfonate (532.2 mg, 2.77 mmol, 1.3 equiv.) at 0°C and then stirred at 80°C for 16 h. The 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 afford the title compound (190.0 mg, 29% yield) as a brown oil. 1H NMR(400MHz,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).

[0190] Step C: (3R)-1-(2-methoxypropyl)piperidin-3-amine A solution of benzyl N-[(3R)-1-(2-methoxypropyl)-3-piperidyl]carbamate (190.0 mg, 0.62 mmol, 1.0 equiv), Pd(OH) (10 mg, 10% palladium on carbon), and Pd / C (10 mg, 10% palladium on carbon) in methanol (6 mL) was stirred at 25 °C for 2 h under H at 1100 mmHg. Upon completion of the reaction, the suspension was filtered through a Celite pad, and the pad was washed with MeOH (5 mL × 3). The combined filtrate was concentrated in vacuo to give the desired product as a yellow oil, which was used directly without further purification to give the title compound (75.0 mg, 70% yield) as a yellow oil. 1 H NMR(400MHz,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,1 H),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).

[0191] Step D: 6-chloro-N-[(3R)-1-(2-methoxypropyl)-3-piperidyl]-5-methyl-1,2,4-triazin-3-amine To a solution of (3R)-1-(2-methoxypropyl)piperidin-3-amine (75.0 mg, 0.44 mmol, 1.0 equiv.), DIEA (0.14 mL, 0.78 mmol, 1.8 equiv.) in 1,4-dioxane (1 mL), 3,6-dichloro-5-methyl-1,2,4-triazine (71.4 mg, 0.44 mmol, 1.0 equiv.) was added and stirred at 25 °C for 16 h. Upon completion of the reaction, the reaction mixture was quenched with HO (10 mL), extracted with EtOAc (20 mL × 2), and the organic phase was washed with brine (10 mL × 2), dried over anhydrous NaSO, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO, petroleum ether:ethyl acetate, 1:1) to afford the title compound (36.0 mg, 28% yield) as a dark green oil. LCMS: m / z 300.2 [M+H] + ,ESI pos.

[0192] Step E: 5-[5-methyl-3-[[(3R)-1-(2-methoxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol To a solution of 6-chloro-N-[(3R)-1-(2-methoxypropyl)-3-piperidyl]-5-methyl-1,2,4-triazine-3-amine (18.0 mg, 0.06 mmol, 1.0 equiv.), 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 equiv.; Example 1, Step C), and CsF (6.98 mg, 0.12 mmol, 2.0 equiv.) in 1 mL of 1,4-dioxane / 0.2 mL of water was added XPhos Pd G3 (10.17 mg, 0.01 mmol, 0.2 equiv.) under N2 at 25 °C, followed by stirring at 95 °C for 2 h. Upon completion of the reaction, the reaction mixture was cooled to room temperature. The combined filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (instrument: ACSWH-GX-M, column: Waters Xbridge 150*25mm*5um, mobile phase: A (for H2O) (0.1% NH4HCO3) and B (for acetonitrile), gradient: B 28% -58% in 10 min, linear flow rate: 30 mL / min, column temperature: RT, wavelength: 220 nm / 254 nm) and lyophilized to give the title compound (9.58 mg, 36% yield) as a yellow oil. LCMS: m / z 400.2 [M+H] + ,ESI pos.

[0193] Example 8: 5-[5-methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0194] Step A: Benzyl (R)-(1-(oxetan-3-yl)piperidin-3-yl)carbamate To a solution of benzyl (R)-piperidin-3-ylcarbamate (CAS No. 478646-32-1; 1.0 g, 4.27 mmol, 1.0 equiv.) in DCE (10 mL), oxetan-3-one (CAS No. 6704-31-0; 0.370 g, 5.12 mmol, 1.2 equiv.) was added and stirred at 20 °C for 1 h. NaBH(OAc) (1.18 g, 5.55 mmol, 1.3 equiv.) was then added and stirred at 20 °C for 1 h. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic phase was 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 column chromatography on silica gel (hexane / EtOAc, 1:0 to 0:1, then EtOAc / MeOH, 1:0 to 5:1) to give the title compound (1.20 g, 97% yield) as a white solid. 1 H NMR(400MHz,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).

[0195] Step B: (R)-1-(oxetan-3-yl)piperidin-3-amine To a solution of benzyl (R)-(1-(oxetan-3-yl)piperidin-3-yl)carbamate (200 mg, 0.69 mmol, 1.0 equiv) in methanol (4 mL) was added Pd / C (10.0 mg, 10% palladium on carbon) under N. After purging with H three times, the mixture was stirred at 20° C. for 1 h at 1100 mmHg. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give the title compound (100 mg, 93% yield) as a yellow oil. 1H NMR(400MHz,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).

[0196] Step C: (R)-6-chloro-5-methyl-N-(1-(oxetan-3-yl)piperidin-3-yl)-1,2,4-triazin-3-amine To a solution of DIPEA (0.157 g, 1.22 mmol, 2.0 equiv.), (R)-1-(oxetan-3-yl)piperidin-3-amine (0.100 g, 0.64 mmol, 1.05 equiv.) in 1,4-dioxane (1 mL), 3,6-dichloro-5-methyl-1,2,4-triazine (CAS No. 132434-82-3, 0.100 g, 0.61 mmol, 1.0 equiv.) was added and stirred at 20 °C for 12 hours. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic phase was 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 column chromatography on silica gel (hexane / EtOAc, 1:0 to 0:1) to give the title compound (0.03 g, 0.11 mmol, 17% yield) as a yellow solid. 1 H NMR(400MHz,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).

[0197] Step D: 5-[5-methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol 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 equiv.), 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 equiv.; Example 1, Step C), KF (0.015 g, 0.26 mmol, 5.0 equiv.) in 1,4-dioxane (1 mL) and water (0.2 mL) was added XPhos Pd G3 (9 mg, 0.01 mmol, 0.2 equiv.) under N2 at 25 °C, followed by stirring at 90 °C for 4 h. The reaction mixture was cooled to room temperature. The mixture was 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 10 μm; conditions: water (NH4HCO3)-ACN start B 21 end B 51; gradient time (min) 10 100% B retention time (min) 2; flow rate (mL / min) 25) to give the title compound (4.35 mg, 20% yield) as a yellow solid. LCMS: m / z 384.1 [M+H] + ,ESI pos.

[0198] Example 9: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoic acid [ka]

[0199] Step A: tert-butyl-(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylic acid To a solution of 3,6-dichloro-5-methyl-1,2,4-triazine (CAS No. 132434-82-3, 520 mg, 3.17 mmol, 1.0 equiv.) in 1,4-dioxane (10 mL) was added (3R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester (CAS No. 188111-79-7, 762 mg, 3.81 mmol, 1.2 equiv.) and N,N-diisopropylethylamine (472 mg, 0.97 mL, 3.66 mmol, 1.80 equiv.) at ambient temperature. The reaction mixture was stirred at 23 °C for 48 h. The reaction mixture was quenched with half-saturated NaHCO solution (100 mL) and extracted with ethyl acetate (2 × 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 (815 mg, 74% yield) as a pale yellow gum. LCMS: m / z 326.0 ([{35Cl}MH] - ), 328.0([{37Cl}MH] - ),ESI neg.

[0200] 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 A sealable tube was placed under argon with a mixture of tert-butyl (3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidine-1-carboxylic acid (Example 9, Step A) (165 mg, 0.48 mmol, 1.0 equiv.), 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 equiv.), and cesium carbonate (467 mg, 1.43 mmol, 3.0 equiv.) in 1,4-dioxane (4 mL) and water (1 mL), and finally XPhos Pd G3 gt (61 mg, 0.72 mmol, 0.15 equiv.) was added. The reaction mixture was stirred in the sealed tube at 95 °C for 4 hours. 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 (152 mg, 74% yield) as a pale yellow foam. LCMS: m / z 428.3 [M+H] + ,ESI pos.

[0201] Step C: 5-[5-methyl-3-[[(3R)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; dihydrochloride 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 equiv) in CHCl (6 mL) and MeOH (2 mL) at +10° C. was added dropwise a 4 M solution of HCl in 1,4-dioxane (1.07 g, 0.89 mL, 3.56 mmol, 10 equiv). The reaction mixture was stirred at 23° C. for 16 h. The reaction mixture was evaporated to dryness to afford the title compound (145 mg, 97% yield) as a pale yellow foam. LCMS: m / z 326.1 [M−H]- ,ESI neg.

[0202] Step D: Methyl 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoate To a suspension of 5-[5-methyl-3-[[(3R)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; dihydrochloride salt (Example 9, Step C) (145 mg, 0.34 mmol, 1.0 equiv) in dichloromethane (7 mL) was added methyl 4-oxobutanoate (CAS No. 13865-19-5, 111 mg, 0.10 mL, 0.86 mmol, 2.50 equiv) followed by sodium acetate ((CAS No. 127-09-3, 71 mg, 0.86 mmol, 2.5 equiv) and sodium triacetoxyborohydride ( A solution of 131 mg of CAS No. 56553-60-7 (0.62 mmol, 1.80 equiv.) was added. The reaction mixture was stirred at 0°C for 5 minutes and at room temperature for 3 hours. Saturated NaHCO3 solution (30 ml) was added to the reaction mixture and 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, 0% to 50% gradient in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (89 mg, 57% yield) as a yellow foam. MS: m / z 426.2 [M−H] - ,ESI neg.

[0203] Step E: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoic acid 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-piperidyl]butanoate (Example 9, Step D) (88 mg, 0.20 mmol, 1.0 equiv.) in tetrahydrofuran (1 mL) and methanol (0.5 mL) was added 1 M aqueous LiOH (0.59 mL, 0.59 mmol, 3.0 equiv.) dropwise at ambient temperature. The yellow reaction solution was stirred at 23° C. for 16 hours. The reaction mixture was neutralized with 5% citric acid and set to pH=4. The aqueous phase was saturated with solid sodium chloride and extracted with dichloromethane (4×30 mL). The combined organic extracts were dried over sodium sulfate, filtered, and the solvent was evaporated in vacuo to give the title compound (38 mg, 45% yield) as a pale yellow solid. MS: m / z 412.1 [M−H] - ,ESI neg.

[0204] Example 10: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N-methyl-butanamide [ka] To a solution of the above-mentioned 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoic acid (Example 9, Step E) (6.4 mg, 0.015 mmol, 1.00 equiv.) and N,N-dimethylformamide (0.4 mL) at ambient temperature was added N,N-diisopropylethylamine (5.7 mg, 0.008 mL, 0.044 mmol, 3.00 equiv.), followed by HATU (CAS No. 148893-10-1, 7.30 mg, 0.019 mmol, 1.30 equiv.). The yellow solution was stirred for 2 minutes, and then monomethylamine hydrochloride (CAS No. 593-51-1, 1.50 mg, 0.022 mmol, 1.50 equiv.) was added. The reaction mixture was stirred at 23°C for 60 hours. 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 um, 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 (3.3 mg, 50% yield) as a pale yellow solid. LCMS: m / z 427.2 [M+H] + ,ESI pos.

[0205] Example 11: 5-[5-methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0206] Step A: Methyl 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]propanoate To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidyl]amine 1:2 hydrochloride (Example 1, Step E) (171 mg, 0.455 mmol, 1.0 equiv.) in tetrahydrofuran, extra-dry (2 mL) and N,N-dimethylformamide, extra-dry (2 mL), N,N-diisopropylethylamine (294 mg, 0.387 mL, 0.0023 mmol, 5.0 equiv.) was added, followed by the dropwise addition of 3-bromopropionic acid methyl ester (CAS No. 3395-91-3, 114 mg, 0.075 mL, 0.683 mmol, 1.5 equiv.). The reaction mixture was stirred at 60° C. for 20 hours. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (2×40 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 give the title compound (108 mg, 72% yield) as a colorless oil. LCMS: m / z 314.1 [M+H] + ,ESI pos.

[0207] Step B 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]-N-(2,2-dimethoxyethyl)propanamide To a solution of 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidino]propionic acid methyl ester (Example 11, Step A) (108 mg, 0.33 mmol, 1.0 equiv.) in tetrahydrofuran (2 mL) and methanol (1 mL) was added 1 M aqueous lithium hydroxide solution (0.425 mL, 0.425 mmol, 1.3 equiv.) dropwise. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was then concentrated. The residue was dissolved twice in toluene, concentrated, and dried under high vacuum at 50° C. for 1 hour. The residue was dissolved in extra-dry N,N-dimethylformamide (3 mL) and N-ethyldiisopropylamine (CAS No. 7087-98-5, 211.3 mg, 0.278 mL, 1.6 mmol, 5.0 equiv.) and HATU (CAS No. 148893-10-1, 187 mg, 0.49 mmol, 1.5 equiv.) were added at ambient temperature. After stirring for 2 minutes, 2,2-dimethoxyethylamine (62 mg, 0.064 mL, 0.589 mmol, 1.8 equiv.) was added. The reaction mixture was stirred at 23 °C for 3 hours. The reaction mixture was quenched with water (15 mL) and extracted with ethyl acetate (2 × 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% (dichloromethane:methanol:NH4OH 110:10:1) in dichloromethane) to give the title compound (137 mg, 97% yield) as a light brown oil. LCMS: m / z 387.3 [M+H] + ,ESI pos.

[0208] Step C: 6-chloro-5-methyl-N-[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidyl]-1,2,4-triazin-3-amine A mixture of 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidino]-N-(2,2-dimethoxyethyl)propionamide (Example 11, Step B) (137 mg, 0.32 mmol, 1.0 equiv.) and Eaton's reagent (CAS No. 39394-84-8, 3.79 g, 2.5 mL, 16 mmol, 50 equiv.) was stirred at 100° C. overnight. The reaction mixture was cooled to room temperature, saturated aqueous NaHCO (100 mL) (strong gas evolution; pH checked: approx. 7) was added dropwise, and the mixture was extracted with dichloromethane (3×60 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% (dichloromethane:methanol:NH4OH 110:10:1) in dichloromethane) to give the title compound (34 mg, 31% yield) as a colorless oil. LCMS: m / z 323.2 [M+H] + ,ESI pos.

[0209] Step D: 5-[5-methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidyl]amine (Example 11, Step C) (34 mg, 0.1 mmol, 1.0 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)coumaran-4-ol (Example 1, Step C) (39 mg, 0.15 mmol, 1.5 equiv.), and cesium carbonate (88 mg, 0.3 mmol, 3.0 equiv.) in 1,4-dioxane (1.6 mL) and water (0.4 mL) was placed under argon and XPhos Pd g3 gt (12.7 mg, 0.015 mmol, 0.15 equiv.) was added. The reaction mixture was stirred in a sealed tube at 90 °C for 3 hours. The reaction mixture was cooled to room temperature, quenched with water (20 mL), saturated aqueous NH4Cl (20 mL) was added, 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 chromatography on silica gel (gradient 0% to 80% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (24 mg, 54% yield) as a light brown foam. LCMS: m / z 423.3 [M+H] + ,ESI pos.

[0210] Example 12: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N,N-dimethyl-butanamide [ka] To a solution of 4-[(3R)-3-[[6-(4-hydroxycoumaran-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 equiv.) and N,N-dimethylformamide (0.3 mL) was added N,N-diisopropylethylamine (60.5 mg, 0.08 mL, 0.468 mmol, 5.0 equiv.) at ambient temperature, followed by HATU (CAS No. 148893-10-1, 55.0 mg, 0.14 mmol, 1.5 equiv.). The yellow solution was stirred for 5 minutes, followed by the addition of dimethylamine hydrochloride (CAS No. 506-89-2, 13.7 mg, 0.168 mmol, 1.8 equiv.). 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 chromatography on silica gel (gradient 0% to 50% (dichloromethane:methanol:NH4OH 110:10:1) in dichloromethane). The product was then purified by preparative HPLC (column: Gemini NX, 12 nm, 5 um, 100 x 30 mm; conditions: ACN / water + 0.1% TEA; gradient: ACN in water, run time 4.5 min) to give the title compound (9 mg, 21% yield) as a pale yellow lyophilized solid. LCMS: m / z 441.5 [M+H] + ,ESI pos.

[0211] Example 13: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-1-pyrrolidin-1-yl-butan-1-one [ka] To a solution of 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (Example 9, Step E) (30.0 mg, 0.065 mmol, 1.0 equiv.) and N,N-dimethylformamide (0.265 mL) was added N,N-diisopropylethylamine (42.2 mg, 0.056 mL, 0.327 mmol, 5.0 equiv.) followed by HATU (CAS No. 148893-10-1, 38.4 mg, 0.098 mmol, 1.5 equiv.) at ambient temperature. The yellow solution was stirred for 5 minutes, and then pyrrolidine (CAS No. 123-75-1, 8.4 mg, 0.0097 mL, 0.118 mmol, 1.8 equiv.) was added. The reaction mixture was stirred for 16 hours at 23° C. Then, HATU (38.4 mg, 0.098 mmol, 1.5 equiv.), N,N-diisopropylethylamine (42.2 mg, 0.059 mL, 0.327 mmol, 5.0 equiv.), and 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (30 mg, 0.065 mmol, 1.0 equiv.) were added to the reaction mixture, and stirring was continued for 48 hours at 23° C. 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 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 (7.3 mg, 22% yield) as a yellow solid. LCMS: m / z 467.4 [M+H] + ,ESI pos.

[0212] Example 14: 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-piperidyl]-N-(2-hydroxyethyl)-N-methylbutanamide [ka] To a solution of 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (Example 9, Step E) (30 mg, 0.065 mmol, 1.0 equiv.) and N,N-dimethylformamide (0.265 mL) was added extra dry N,N-diisopropylethylamine (42.2 mg, 0.056 mL, 0.327 mmol, 5.0 equiv.) at ambient temperature, followed by HATU (CAS No. 148893-10-1, 38.4 mg, 0.098 mmol, 1.5 equiv.). The yellow solution was stirred for 2 minutes, and then 2-(methylamino)ethanol (CAS No. 109-83-1, 8.83 mg, 0.0094 mL, 0.118 mmol, 1.8 equiv.) was added. The reaction mixture was stirred at 23° C. for 16 hours. HATU (38.4 mg, 0.098 mmol, 1.5 equiv.), N,N-diisopropylethylamine (42.2 mg, 0.0556 mL, 0.327 mmol, 5.0 equiv.), and 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (Example 9, Step E) (30 mg, 0.065 mmol, 1.0 equiv.) 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 (6 mg, 18% yield) as a yellow solid. LCMS: m / z 471.4 [M+H] + ,ESI pos.

[0213] Example 15: 1-(3-Hydroxypyrrolidin-1-yl)-4-[rac-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butan-1-one; Formic acid [ka] To a solution of 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (Example 9, Step E) (34 mg, 0.074 mmol, 1.0 equiv) and N,N-dimethylformamide, extra dry (0.3 mL) at ambient temperature was added N,N-diisopropylethylamine (47.8 mg, 0.063 uL, 0.37 mmol, 5.0 equiv), followed by HATU (CAS No. 148893-10-1, 43.5 mg, 0.111 mmol, 1.5 equiv). The yellow solution was stirred for 2 minutes, then DL-pyrrolidin-3-ol (CAS No. 40499-83-0, 11.6 mg, 0.011 mL, 0.133 mmol, 1.8 equiv.) was added. The reaction mixture was stirred at 23° C. for 60 hours. 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 um, 100×30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: ACN in water, run time 4.5 min) to give the title compound (11 mg, 27% yield) as a light brown foam. LCMS: m / z 483.5 [M+H] + ,ESI pos.

[0214] Example 16: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-1-morpholinobutan-1-one [ka] To a solution of 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyric acid (Example 9, Step E) (40 mg, 0.097 mmol, 1.0 equiv.) and N,N-dimethylformamide (2.5 mL) was added N,N-diisopropylethylamine (62.5 mg, 0.082 mL, 0.484 µmol, 5.0 equiv.) followed by HATU (CAS No. 148893-10-1, 55.2 mg, 0.145 mmol, 1.5 equiv.) at ambient temperature. The yellow solution was stirred for 4 minutes, followed by the addition of morpholine (CAS No. 110-91-8, 15.2 mg, 0.015 mL, 0.174 mmol, 1.8 equiv.). After 24 hours, N,N-diisopropylethylamine (62.5 mg, 0.082 mL, 0.484 mmol, 5.0 equiv) and HATU (55.2 mg, 0.145 mmol, 1.5 equiv) were added again. After stirring for 4 minutes, morpholine (15.2 mg, 0.015 mL, 0.174 mmol, 1.8 equiv) was added again. The reaction mixture was stirred at 23°C for a total of 45 hours. The reaction mixture was quenched with 2 mL of water. The reaction mixture was extracted with water (approximately 10 mL) and ethyl acetate (approximately 15 mL), and then the aqueous layer was back-extracted with ethyl acetate (approximately 15 mL). The organic layer was washed with brine (approximately 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 um, 100 x 30 mm; Conditions: ACN / water + 0.1% TEA; Gradient: ACN in water, Run time 4.5 min) to give the title compound (4 mg, 8% yield) as a pale yellow lyophilized solid. LCMS: m / z 483.5 [M+H] + ,ESI pos.

[0215] Example 17: 5-[5-methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0216] Step A: 6-chloro-5-methyl-N-[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidyl]-1,2,4-triazin-3-amine To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidyl]amine (Example 1, Step E) (250 mg, 1.1 mmol, 1.0 equiv.) in tetrahydrofuran, extra dry (4 mL) and N,N-dimethylformamide, extra dry (4 mL) was added N,N-diisopropylethylamine (568 mg, 0.7467 mL, 4.4 mmol, 4.0 equiv.), followed by 1-(2-bromoethyl)pyrazole hydrobromide (CAS No. 1955531-53-9, 443.7 mg, 1.65 mmol, 1.5 equiv.). The reaction mixture was stirred at 60° C. for 20 hours. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2×60 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 (231 mg, 62% yield) as a light brown oil. LCMS: m / z 322.3 ([{35Cl}M+H] + ), 324.2([{37Cl}M+H] + ),ESI pos.

[0217] Step B: 5-[5-methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidyl]amine (Example 17, Step A) (113 mg, 0.351 mmol, 1.0 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)coumaran-4-ol (Example 1, Step C) (128.9 mg, 0.492 mmol, 1.4 equiv.), and cesium carbonate (343 mg, 1.05 mmol, 3.0 equiv.) in 1,4-dioxane (4 mL) and water (1 mL) was placed under argon. Finally, XPhos Pd g3 gt (44.6 mg, 0.053 mmol, 0.15 equiv.) 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 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% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (89 mg, 57% yield) as a yellow foam. LCMS: m / z 422.4 [M+H] + ,ESI pos.

[0218] Examples 18 and 19: 5-[5-methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; Formic acid and 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanenitrile [ka] and [ka]

[0219] Step A: 4-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]butanenitrile To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidyl]amine (Example 1, Step E) (400 mg, 1.76 mmol, 1.0 equiv.) in extra-dry tetrahydrofuran (6 mL) and extra-dry N,N-dimethylformamide (6 mL) was added N,N-diisopropylethylamine (567 mg, 0.747 mL, 4.4 mmol, 2.5 equiv.), followed by the dropwise addition of 4-bromobutyronitrile (CAS No. 5332-06-9, 390 mg, 0.26 mL, 2.64 mmol, 1.5 equiv.). The reaction mixture was stirred at 60° C. for 16 hours. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2×60 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 (391 mg, 72% yield) as a light brown oil. LCMS: m / z 295.2 ([{35Cl}M+H] + ), 297.2([{37Cl}M+H] + ),ESI pos.

[0220] Step B: 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanenitrile A mixture of 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 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)coumaran-4-ol (Example 1, Step C) (187 mg, 0.712 mmol, 1.4 equiv.), and cesium carbonate (498 mg, 1.53 mmol, 3.0 equiv.) in 1,4-dioxane (4 mL) and water (1 mL) was placed under argon, and finally XPhos Pd g3 gt (65 mg, 0.076 mmol, 0.15 equiv.) 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 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% 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 afford the title compound (145 mg, 69% yield) as a yellow foam. LCMS: m / z 395.2 [M+H] + ,ESI pos.

[0221] Step C: 5-[5-methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; formic acid To a solution of 4-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]butyronitrile (Example 18, Step B) (135 mg, 0.343 mmol, 1.0 equiv.) in N,N-dimethylformamide was added extra-dry (1.45 mL) sodium azide (CAS No. 26628-22-8, 100 mg, 1.54 mmol, 4.5 equiv.), followed by L-proline (CAS No. 147-85-3, 11.8 mg, 0.103 mmol, 0.3 equiv.). The reaction mixture was stirred in a sealed tube at 115° C. for 16 h. After 16 hours, additional sodium azide (100 mg, 1.54 mmol, 4.5 equiv.) and L-proline (11.8 mg, 0.103 mmol, 0.3 equiv.) were added, and stirring was continued at 120 °C for another 60 hours. The reaction mixture was quenched with water (5 mL) and extracted with ethyl acetate (2 × 30 mL). The aqueous phase was evaporated, and the solid residue was triturated in DCM / MeOH 9:1, some sodium sulfate was added, and then filtered. The product was then purified by preparative HPLC (column: YMC-Triart C18, 12 nm, 5 μm, 100 × 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient 5-50% ACN, run time 4.5 min) to afford the title compound (51 mg, 29% yield) as a light brown lyophilized solid. LCMS: m / z 438.4 [M+H] + ,ESI pos.

[0222] Example 20: 5-[5-methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0223] Step A: 6-chloro-5-methyl-N-[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidyl]-1,2,4-triazin-3-amine To a solution of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidyl]amine 1:2 hydrochloride (Example 1, Step B) (67 mg, 0.178 mmol, 1.0 equiv.) in dichloromethane was added extra-dry (2 mL) 3-oxazol-2-ylpropionaldehyde (CAS No. 1214937-88-8, 31.2 mg, 0.25 mmol, 1.4 equiv.) at 0° C., followed by sodium acetate (36.6 mg, 0.446 mmol, 2.0 equiv.) and sodium triacetoxyborohydride (CAS No. 56553-60-7, 68 mg, 0.321 mmol, 1.8 equiv.). The reaction mixture was stirred at 0° C. for 5 minutes and at room temperature for 3 hours. To the reaction mixture was added saturated NaHCO3 solution (20 mL) 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 chromatography on silica gel (gradient 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (43 mg, 72% yield) as a light brown oil. LCMS: m / z 337.2 ([{35Cl}M+H] + ), 339.2([{37Cl}M+H] + ),ESI pos.

[0224] Step B: 5-[5-methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidyl]amine (Example 20, Step A) (43 mg, 0.128 mmol, 1.0 equiv.), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)coumaran-4-ol (Example 1, Step C) (55.8 mg, 0.192 mmol, 1.5 equiv.), and cesium carbonate (125 mg, 0.383 mmol, 3.0 equiv.) in 1,4-dioxane (1.6 mL) and water (0.4 mL) was placed under argon, and finally XPhos Pd G3 gt (16 mg, 0.019 mmol, 0.15 equiv.) 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, quenched with water (10 mL) and saturated aqueous NH4Cl (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 chromatography on silica gel (gradient 0% to 50% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (19 mg, 32% yield) as a pale yellow foam. LCMS: m / z 437.3 [M+H] + ,ESI pos.

[0225] Example 21: 5-[5-methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0226] Step A: 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]-1-piperidyl]propanenitrile To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidyl]amine (Example 1, Step E) amine (500 mg, 2.2 mmol, 1.0 equiv.) in extra-dry tetrahydrofuran (8 mL) and extra-dry N,N-dimethylformamide (8 mL), N,N-diisopropylethylamine (710 mg, 0.934 mL, 5.5 mmol, 2.5 equiv.) was added, followed by dropwise addition of 3-bromopropionitrile (CAS No. 2417-90-5, 465 mg, 0.287 mL, 3.3 mmol, 1.5 equiv.). The reaction mixture was stirred at 60° C. for 48 hours. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (2×60 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 (425 mg, 65% yield) as a light brown oil. LCMS: m / z 281.2 ([{35Cl}M+H] + ), 283.2([{37Cl}M+H] + ),ESI pos.

[0227] Step B: 3-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]propanenitrile A mixture of 3-[(3R)-3-[(6-chloro-5-methyl-1,2,4-triazin-3-yl)amino]piperidino]propionitrile (Example 22, Step A, extra dry) (200 mg, 0.712 mmol, 1.0 equiv.) was dissolved in 1,4-dioxane, extra dry (4 mL) and water (1 mL). Then, 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)coumaran-4-ol (Example 1, Step C) (280 mg, 1.07 mmol, 1.5 equiv.), cesium carbonate (696 mg, 2.2 mmol, 3.0 equiv.), and XPhos Pd g3 (90.5 mg, 0.10 mmol, 0.15 equiv.) were added under an argon atmosphere. 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) and 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% in dichloromethane (dichloromethane:methanol:NH4OH 110:10:1)) to afford the title compound (240 mg, 80% yield) as a light brown oil. LCMS: m / z 381.3 [M+H] + ,ESI pos.

[0228] Step C: 5-[5-methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol To a solution of 3-[(3R)-3-[[6-(4-hydroxycoumaran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]piperidino]propionitrile (Example 22, Step B) (100 mg, 0.237 mmol, 1.0 equiv.) in N,N-dimethylformamide was added extra-dry (1 mL) L-proline (CAS No. 147-85-3, 8.2 mg, 0.071 μmol, 0.30 equiv.), followed by sodium azide (CAS No. 26628-22-8, 69 mg, 1.06 mmol, 4.5 equiv.). The reaction mixture was stirred at 115° C. for 16 hours. After 16 hours, L-proline (8.2 mg, 0.071 mmol, 0.3 equiv.) and sodium azide (69 mg, 1.06 mmol, 4.5 equiv.) were added again. The reaction mixture was stirred at 115° C. for 48 hours. The reaction mixture was quenched with water (approx. 5 mL) and then extracted with ethyl acetate (approx. 25 mL). The product was in the aqueous phase. The aqueous layer was back-extracted four times with ethyl acetate (approx. 25 mL). The aqueous layer was concentrated in vacuo. The crude product was then purified by preparative HPLC (column: YMC-Triart C 18 Purification by chromatography (HPLC, 12 nm, 5 μm, 100 × 30 mm; conditions: ACN / water + 0.1% HCOOH; gradient: 5 to 50% ACN, run time 4.5 min) afforded the title (3 mg, 3% yield) as a light brown lyophilized solid. LCMS: m / z 242.3 [M+H] + ,ESI pos.

[0229] Example 22: 5-[5-methyl-3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol [ka]

[0230] Step A: 6-chloro-5-methyl-N-[(3R)-1-(3-methylsulfonylpropyl)-3-piperidyl]-1,2,4-triazin-3-amine To a mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-3-piperidyl]amine (Example 1, Step E) (100 mg, 0.439 mmol, 1.0 equiv.) in extra-dry tetrahydrofuran (1.6 mL) and extra-dry N,N-dimethylformamide (1.6 mL) was added N,N-diisopropylethylamine (142.0 mg, 0.187 mL, 1.1 mmol, 2.5 equiv.), followed by the dropwise addition of 1-bromo-3-mesylpropane (CAS No. 859940-73-1, 132.5 mg, 0.659 mmol, 1.5 equiv.). The reaction mixture was stirred at 60° C. for 12 hours. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (2×40 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 give the title compound (111 mg, 65% yield) as an orange oil. LCMS: m / z 348.2 [M+H] + ,ESI pos.

[0231] Step B: 5-[3-[[(3R)-1-(3-mesylpropyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]coumaran-4-ol A mixture of (6-chloro-5-methyl-1,2,4-triazin-3-yl)-[(3R)-1-(3-mesylpropyl)-3-piperidyl]amine (Example 23, Step A) (108 mg, 0.310 mmol, 1.0 equiv), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)coumaran-4-ol (Example 1, Step C) (144.67 mg, 0.497 mmol, 1.6 equiv), cesium carbonate (305 mg, 0.936 mmol, 3.0 equiv) and XPhos Pd g3 (39.4 mg, 0.047 mmol, 0.15 equiv) 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 half-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 crude product was purified by flash chromatography (Si-amine, 12 g, gradient 0% to 10% methanol in DCM) to afford the title compound (108 mg, 70% yield) as a yellow solid. LCMS: m / z 448.3 [M+H] + ,ESI pos.

[0232] Example A The compounds of formula I can be used in a manner known per se as active ingredient to produce tablets of the following composition: Per tablet Active ingredient 200mg Microcrystalline cellulose 155mg Cornstarch 25mg Talc 25mg Hydroxypropyl methylcellulose 20mg 425mg

[0233] Example B The compounds of formula I can be used in a manner known per se as active ingredient to prepare capsules of the following composition: Per capsule Active ingredient 100.0mg Cornstarch 20.0mg Lactose 95.0mg Talc 4.5mg Magnesium stearate 0.5mg 220.0mg

Claims

1. Formula I: 【Chemistry 31】 [In the formula, R １ and R ５ , and the atoms to which they are attached, i. a 4-6 membered heterocycle containing a single O heteroatom, or ii. Forms any of 4- to 5-membered cycloalkyl rings; R ２ and R ３ is selected from H and alkyl, R ２ or R ３ Only one of may be H or alkyl; R ４ Below: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, cycloalkyl substituted with hydroxy or alkoxy; v. cycloalkylalkyl substituted with hydroxyl, alkoxy or —COOH; vi. a 4-6 membered heterocycle containing a single O heteroatom; vii. (CH ２ ) ｎ C(O)OH, (CH ２ ) ｎ C(O)OCH ３ , or (CH ２ ) ｎ C(O)NHCH ３ wherein n is 2 or 3; viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is tetrazole, oxadiazole, or oxazole heteroaryl; ix. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; x. -(CH ２ ) ｎ S (O) ２ CH ３ or (CH ２ ) ｎ CN, where n is 3; xi. -(CH ２ ) ｎ C(O)NR'R'' where n is 3 and R' and R'' are both CH ３ or R' is CH ３ and R″ is hydroxylalkyl, or R′ and R″ together with the N to which they are attached represent a. a 5-membered heterocycle, said heterocycle being optionally substituted with OH, or b. A 6-membered heterocycle further containing one O heteroatom (These form either Selected from: R ６ is H or —OH, R ４ is R ６ can be alkyl only if is —OH] and pharmaceutically acceptable salts thereof.

2. R １ and R ５ and the atoms to which they are attached form a 5-membered heterocycle containing a single O heteroatom.

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

4. R ４ but the following: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. a 4-membered heterocycle containing a single O heteroatom; vi. (CH ２ ) ｎ C(O)OH, (CH ２ ) ｎ C(O)OCH ３ , or (CH ２ ) ｎ C(O)NHCH ３ wherein n is 3; vii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazoleheteroaryl; viii. heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; ix. -(CH ２ ) ｎ S (O) ２ CH ３ or (CH ２ ) ｎ CN, where n is 3; x. -(CH ２ ) ｎ C(O)NR'R'' where n is 3 and R' and R'' are both CH ３ or R' is CH ３ and R″ is hydroxylalkyl, or R′ and R″ together with the N to which they are attached represent a. a 5-membered heterocycle, said heterocycle being optionally substituted with OH, or b. A 6-membered heterocycle further containing one O heteroatom (These form either Selected from: R ４ is R ６ may be alkyl only if —OH; The compound according to any one of claims 1 to 3.

5. R ４ but the following: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. a 4-membered heterocycle containing a single O heteroatom; vi. (CH ２ ) ｎ C(O)OH, (CH ２ ) ｎ C(O)OCH ３ , or (CH ２ ) ｎ C(O)NHCH ３ , (wherein n is 3), vii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is an oxazole heteroaryl. Selected from: R ４ is R ６ may be alkyl only if —OH; The compound according to any one of claims 1 to 3.

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

7. R ４ but, i. cycloalkyl substituted with hydroxy or alkoxy, and ii. Cycloalkylalkyl substituted with hydroxy or alkoxy The compound according to any one of claims 1 to 3, selected from:

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

9. 2. The compound of claim 1 , R １ and R ５ and the atoms to which they are attached form a 5-membered heterocycle containing a single O heteroatom; R ２ is H and R ３ is methyl; R ４ Below: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. a 4-membered heterocycle containing a single O heteroatom; vi. (CH ２ ) ｎ C(O)OH, (CH ２ ) ｎ C(O)OCH ３ , or (CH ２ ) ｎ C(O)NHCH ３ wherein n is 3; vii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is oxazoleheteroaryl; viii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is pyrazole; ix. -(CH ２ ) ｎ S (O) ２ CH ３ or (CH ２ ) ｎ CN, where n is 3; x. -(CH ２ ) ｎ C(O)NR'R'' where n is 3 and R' and R'' are both CH ３ or R' is CH ３ and R″ is hydroxylalkyl, or R′ and R″ together with the N to which they are attached represent a. a 5-membered heterocycle, said heterocycle being optionally substituted with OH, or b. A 6-membered heterocycle further containing one O heteroatom (These form either Selected from: R ６ is H or —OH, R ４ is R ６ may be alkyl only if —OH; Compounds and pharmaceutically acceptable salts.

10. 2. The compound of claim 1 , R １ and R ５ and the atoms to which they are attached form a 5-membered heterocycle containing a single O heteroatom; R ２ is H and R ３ is methyl; R ４ Below: i. alkyl, ii. hydroxyalkyl, iii. alkoxyalkyl, iv. cycloalkylalkyl substituted with hydroxy; v. a 4-membered heterocycle containing a single O heteroatom; vi. (CH ２ ) ｎ C(O)OH, (CH ２ ) ｎ C(O)OCH ３ , or (CH ２ ) ｎ C(O)NHCH ３ wherein n is 3; vii. Heteroarylalkyl, wherein the heteroaryl in the heteroarylalkyl is an oxazole heteroaryl. Selected from: R ６ is H or —OH, R ４ is R ６ may be alkyl only if —OH; Compounds and pharmaceutically acceptable salts.

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

12. The compound is 5-[5-methyl-3-[[rac-(3R)-1-(2-hydroxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-[(1-hydroxycyclopropyl)methyl]-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidyl]amino]-5-methyl-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; Formate salts; 5-[3-[[(3R)-1-(2-methoxyethyl)-3-piperidyl]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-[[rac-(3R)-1-(2-methoxypropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-methyl-3-[[(3R)-1-(oxetan-3-yl)-3-piperidyl]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-piperidyl]butanoic acid; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]; Methyl 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butanoate; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N-methyl-butanamide; 5-[5-methyl-3-[[(3R)-1-(2-oxazol-2-ylethyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; and pharmaceutically acceptable salts thereof.

13. The compound is 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N,N-dimethyl-butanamide; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-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-piperidyl]-N-(2-hydroxyethyl)-N-methyl-butanamide; 1-(3-hydroxypyrrolidin-1-yl)-4-[rac-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butan-1-one; Formic acid; 1-(3-hydroxypyrrolidin-1-yl)-4-[rac-(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]butan-1-one; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-1-morpholino-butan-1-one; 5-[5-methyl-3-[[(3R)-1-(2-pyrazol-1-ylethyl)-3-piperidyl]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-piperidyl]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-piperidyl]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-piperidyl]butanenitrile; 5-[5-methyl-3-[[(3R)-1-(3-oxazol-2-ylpropyl)-3-piperidyl]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-piperidyl]-N,N-dimethyl-butanamide; formic acid'' 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-N,N-dimethyl-butanamide; formic acid; 5-[5-methyl-3-[[(3R)-1-[2-(1H-tetrazol-5-yl)ethyl]-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; 5-[5-methyl-3-[[(3R)-1-(3-methylsulfonylpropyl)-3-piperidyl]amino]-1,2,4-triazin-6-yl]-2,3-dihydrobenzofuran-4-ol; and pharmaceutically acceptable salts thereof.

14. 2. The compound of 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-piperidyl]butanoic acid or a pharmaceutically acceptable salt thereof.

15. The compound is 5-[5-methyl-3-[[(3R)-1-[3-(1H-tetrazol-5-yl)propyl]-3-piperidyl]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-piperidyl]-N,N-dimethyl-butanamide; 4-[(3R)-3-[[6-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-5-methyl-1,2,4-triazin-3-yl]amino]-1-piperidyl]-1-pyrrolidin-1-yl-butan-1-one; and pharmaceutically acceptable salts thereof.

16. A compound according to any one of claims 1 to 15 for use as a therapeutically active substance.

17. 16. A compound according to any one of claims 1 to 15 for use in the treatment or prevention of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15 and a therapeutically inert carrier.

19. 16. Use of a compound according to any one of claims 1 to 15 for the treatment or prevention of a disease, disorder or condition, wherein said disease, disorder or condition is responsive to NLRP3 inhibition.

20. A compound according to any one of claims 1 to 15 for use in the treatment or prevention of a cardiovascular disease, disorder or condition.

21. 20. Use of a compound according to any one of claims 1 to 15 in the treatment or prevention of a cardiovascular disease, disorder or condition.

22. 20. Use of a compound according to any one of claims 1 to 15 for the preparation of a medicament for treating or preventing a cardiovascular disease, disorder or condition.

23. 16. A method of inhibiting NLRP3, comprising administering an effective amount of a compound according to any one of claims 1 to 15 to inhibit NLRP3.

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

25. The invention as hereinbefore described.

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