NLRP3 inhibitors

Abstract

The present invention relates to compounds of general formula I TIFF2025540258000067.tif35161 [In the formula, R 1 , R 2 , X and W are as described herein, compositions comprising said compounds, and methods of using said compounds.

Description

[Technical Field]

[0001] The present invention relates to organic compounds useful for the treatment and / or prevention in mammals, in particular compounds that modulate NLRP3 inhibition. The present invention relates to a compound of formula I [ka] [In the formula, R 1 is H or alkyl; R 2 teeth, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, optionally substituted with 1-2 substituents selected from alkyl, OH, halo, haloalkyl, hydroxyalkyl, or oxo; ii. -CH2-heterocycle, which is a 5-6 membered heterocycle containing a single O heteroatom; iii. 4-6 membered cycloalkyl optionally substituted with 1-2 substituents independently selected from alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl Selected from; X is -O-, -CH2-, -NH-, or -N(CH3)-; W is the ring system A and B [ka] [ka] (R 3 is H, halo, alkyl, alkoxy, or alkoxyalkyl, where R 1 and R 3 One of them is H and the other is not H; R 4 is halo, alkyl, alkoxy, cyano, haloalkyl, or haloalkoxy; R 5 Is H; or R 4 and R 5 , and the atoms to which they are attached, i) a 4- to 5-membered cycloalkyl optionally substituted with oxo; or ii) a 5-membered heterocycle containing a single O heteroatom optionally substituted with 1 to 2 substituents independently selected from alkyl and halo; Forming; R 6 is halo, haloalkyl, or OH; R 7 is H or F) Novel compounds; and pharmaceutically acceptable salts thereof.

[0002] Furthermore, the present invention includes all racemic mixtures, all their corresponding enantiomers and / or optical isomers. [Background technology]

[0003] The NOD-like receptor (NLR) family pyrin domain-containing protein 3 (NLRP3) inflammasome is a component of the inflammatory process, and its aberrant activity is pathogenic in genetic disorders such as cryopyrin-associated periodic syndromes (CAPS), and complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer's disease, and atherosclerosis.

[0004] 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.

[0005] Caspase-1 cleaves pro-IL-1β and pro-IL-18 to their active forms, which are secreted from cells. Active caspase-1 also cleaves gasdermin-D to induce 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 protein (HMGB1). Caspase-1 also cleaves intracellular IL-1R2, leading to its degradation and enabling the release of IL-1α. In human cells, caspase-1 can also regulate the processing and secretion of IL-37. Several other caspase-1 substrates, such as components of the cytoskeleton and glycolytic pathways, may contribute to caspase-1-dependent inflammation.

[0006] 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.

[0007] 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 synergize to induce IFN-γ production from memory T cells and NK cells, driving Th1 responses.

[0008] 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 key component of the inflammatory process. NLRP3 is also involved in the pathogenesis of several complex diseases, including metabolic disorders such as type 2 diabetes, atherosclerosis, obesity, and gout, among others.

[0009] 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 implicated in several central nervous system conditions, including Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, and brain injury due to 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 be involved in several 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 is involved in the development of liver disease, kidney disease, and aging. Many of these associations are due to the involvement of NLRP3. - / - Although defined using 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.

[0010] 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 in response to 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.

[0011] 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.

[0012] There is a need to provide compounds that have improved pharmacological and / or physiological and / or physicochemical properties and / or that provide useful alternatives to known compounds. Summary of the Invention

[0013] The present invention relates to a compound of formula I [ka] [In the formula, R 1 is H or alkyl; R 2 teeth, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, optionally substituted with 1-2 substituents selected from alkyl, OH, halo, haloalkyl, hydroxyalkyl, or oxo; ii. -CH2-heterocycle, which is a 5-6 membered heterocycle containing a single O heteroatom; iii. 4-6 membered cycloalkyl optionally substituted with 1-2 substituents independently selected from alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl Selected from; X is -O-, -CH2-, -NH-, or -N(CH3)-; W is the ring system A and B [ka] [ka] (R 3 is H, halo, alkyl, alkoxy, or alkoxyalkyl, where R 1 and R 3 One of them is H and the other is not H; R 4 is halo, alkyl, alkoxy, cyano, haloalkyl, or haloalkoxy; R 5 Is H; or R 4 and R 5 , and the atoms to which they are attached, i) a 4- to 5-membered cycloalkyl optionally substituted with oxo; or ii) a 5-membered heterocycle containing a single O heteroatom optionally substituted with 1 to 2 substituents independently selected from alkyl and halo; Forming; R 6 is halo, haloalkyl, or OH; R 7 is H or F) Select from Novel compounds; and pharmaceutically acceptable salts thereof.

[0014] 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 Contains alkyl. 1~6 Examples of alkyl include methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl and pentyl. Particular alkyl groups include methyl and ethyl.

[0015] The term "alkoxy" refers to a group in which R' is C 1~6 represents a group of the 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.

[0016] 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 are methoxymethyl and methoxyethyl.

[0017] 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, and cyclohexyl. A specific example is cyclobutyl.

[0018] The terms "halogen," "halide," and "halo" are used interchangeably herein and refer to fluoro, chloro, bromo, or iodo.

[0019] The term "haloalkyl" refers to C 1~6 C in which at least one hydrogen atom of the alkyl group is replaced by the same or different halogen atom 1~6 represents an alkyl group. Examples of haloalkyl include fluoromethyl, difluoromethyl and trifluoromethyl.

[0020] The term "haloalkoxy" refers to C 1~6 C in which at least one hydrogen atom of the alkoxy group is replaced by the same or different halogen atom 1~6represents an alkoxy group. Examples of haloalkoxy are difluoromethoxy, trifluoromethoxy, difluoroethoxy and trifluoroethoxy.

[0021] The term "heterocycle ring" 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 are azetidinyl, diazepanyl, pyrrolidinyl, tetrahydrofuranyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, and piperazinyl. Examples of polycyclic saturated heterocycles are azaspiroheptanyl, diazaspiroheptanyl, azaspirooctanyl, diazospirooctanyl, diazaspirononanyl, oxaazaspirooctanyl, and oxadiazaspirononanyl. A specific example of a heterocycle is piperidinyl.

[0022] The term "hydroxy" refers to an --OH group.

[0023] 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, hydroxymethylethyl, hydroxymethylpropyl, and dihydroxypropyl. The term "cyano" refers to the group -C≡N.

[0024] The term "oxo" refers to a divalent oxygen atom =O.

[0025] 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 salts formed with hydrochloric acid to produce hydrochloride, dihydrochloride, or trihydrochloride.

[0026] The abbreviation uM means micromolar and is equivalent to the symbol μM.

[0027] The abbreviation uL means microliter and is equivalent to the symbol μL.

[0028] The abbreviation ug stands for microgram and is equivalent to the symbol μg.

[0029] 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.

[0030] According to the Cahn-Ingold-Prelog rules, the asymmetric carbon atom can be of the "R" or "S" configuration.

[0031] Embodiments of the present invention also provide 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.

[0032] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 1 is H or alkyl, and R 3 is H, alkyl or alkoxyalkyl, where R 1 or R 3 One of them is H and the other is not H.

[0033] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 1 is H or alkyl, and R 3 is H or alkyl, where R 1 or R 3 One of the groups is H and the other is alkyl

[0034] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 1 is H and R 3 is alkyl.

[0035] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 2 teeth, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, optionally substituted with alkyl, halo, haloalkyl, hydroxyalkyl, or oxo; ii. -CH2-heterocycle, which is a 5-6 membered heterocycle containing a single O heteroatom; iii. 4-6 membered cycloalkyl optionally substituted with 1-2 substituents independently selected from alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl is selected from.

[0036] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 2 teeth, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, optionally substituted with alkyl or with both alkyl and OH; ii. 4-6 membered cycloalkyl optionally substituted with 1-2 substituents independently selected from alkyl and OH.

[0037] Embodiments of the present invention provide compounds according to Formula I as described herein, wherein R 2 is a 5-6 membered heterocycle containing a single O or a single N heteroatom, the heterocycle being optionally substituted by alkyl or by both alkyl and OH. Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 2 teeth, i. 6-membered heterocycles containing a single N heteroatom substituted with alkyl; ii. -CH2-heterocycle, which is a 5-6 membered heterocycle containing a single O heteroatom; iii. 4-membered cycloalkyl substituted with alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl is selected from.

[0038] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 2 teeth, i. 6-membered heterocycles containing a single N heteroatom substituted with alkyl, and ii. 4-membered cycloalkyl substituted with alkyl and OH is selected from.

[0039] An embodiment of the present invention provides a compound according to Formula I described herein, wherein X is O or —CH 2 —.

[0040] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 7 is H.

[0041] An embodiment of the present invention provides a compound according to formula I described herein, wherein W is a ring system A [ka] is.

[0042] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 5 is H or R 4 and R 5 , and the atoms to which they are attached, i. a 5-membered cycloalkyl ring, or ii. Forms a 5-membered heterocycle containing a single O heteroatom.

[0043] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 5 is H or R 4 and R 5 , and the atoms to which they are attached, i. 4-5 membered cycloalkyl optionally substituted with oxo; or ii. Forms a 5-membered heterocycle containing a single O heteroatom.

[0044] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 5 is H.

[0045] Embodiments of the present invention provide compounds according to Formula I described herein, wherein W is selected from the group consisting of ring systems A, C and D. [ka] [ka] [ka] is selected from During the ceremony, R 3 is H or alkyl, where R 1 and R 3 One of them is H and the other is not H; R 4 is alkyl, cyano, haloalkyl, or haloalkoxy; R 5 is H; R 6 is OH; Y is —CH 2 — or —O—.

[0046] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 4 is halo, cyano, haloalkyl or haloalkoxy.

[0047] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 4 is alkyl, cyano, haloalkyl, or haloalkoxy.

[0048] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R4 is cyano.

[0049] Embodiments of the present invention provide compounds according to Formula I described herein, wherein R 6 is OH.

[0050] An embodiment of the present invention is a compound according to Formula I as described herein. [In the formula, R 1 is H or alkyl; R 2 teeth, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, optionally substituted with alkyl or with both alkyl and OH; ii. 4- to 6-membered cycloalkyl optionally substituted with 1 to 2 substituents independently selected from alkyl and OH Selected from; X is -O- or -CH2-; W is the ring system A, C and D [ka] [ka] [ka] (In the formula, R 3 is H or alkyl, where R 1 and R 3 One of them is H and the other is not H; R 4 is alkyl, cyano, haloalkyl, or haloalkoxy; R 5 is H; R 6 is OH; Y is CH2 or O Select from and pharmaceutically acceptable salts thereof.

[0051] An embodiment of the present invention is a compound according to Formula I as described herein. [In the formula, R 1 is H or alkyl; R 2 is a 5-6 membered heterocycle containing a single O or a single N heteroatom, optionally substituted by alkyl or by both alkyl and OH; X is -O- or -CH2-; W is the ring system C and D [ka] [ka] (In the formula, R 3 is H or alkyl, where R 1 and R 3 One of them is H and the other is not H; R 4 is alkyl, cyano, haloalkyl, or haloalkoxy; R 5 is H; R 6 is OH; Y is CH2 or O Select from and pharmaceutically acceptable salts thereof.

[0052] An embodiment of the present invention is a compound according to Formula I as described herein. (In the formula, R 1 is H or alkyl; R 2 teeth, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, optionally substituted with alkyl, halo, haloalkyl, hydroxyalkyl, or oxo; ii. -CH2-heterocycle, which is a 5-6 membered heterocycle containing a single O heteroatom; iii. 4-6 membered cycloalkyl optionally substituted with 1-2 substituents independently selected from alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl Selected from; X is -O-, -CH2-, -NH-, or -N(CH3)-; W is the ring system A and B [ka] [ka] Selected from; R 3 is H, halo, alkyl, alkoxy, or alkoxyalkyl, where R 1 and R 3 One of them is H and the other is not H; R 4 is halo, alkyl, alkoxy, cyano, haloalkyl, or haloalkoxy; R 5 Is H; or R 4 and R 5 , and the atoms to which they are attached, i. 4-5 membered cycloalkyl optionally substituted with oxo; or ii. A 5-membered heterocycle containing a single O heteroatom optionally substituted with 1 to 2 substituents independently selected from alkyl and halo. Forming; R 6 is halo, haloalkyl, or OH; R 7 is H or F) and pharmaceutically acceptable salts thereof.

[0053] An embodiment of the present invention is a compound according to Formula I as described herein. (In the formula, R 1 is H or alkyl, and R 3 is H or alkyl, where R 1 or R 3 one of which is H and the other is alkyl; R 2 teeth, i. 6-membered heterocycles containing a single N heteroatom substituted with alkyl, and ii. 4-membered cycloalkyl substituted with alkyl and OH Selected from; X is O or -CH2-; W is the ring system A: [ka] and; R 4 is cyano; R 5 is H; R 6 is OH); and pharmaceutically acceptable salts thereof.

[0054] An embodiment of the present invention is a compound according to Formula I as described herein. (In the formula, R 1 is H and R 3 is alkyl; R 2 teeth, i. 6-membered heterocycles containing a single N heteroatom substituted with alkyl, and ii. 4-membered cycloalkyl substituted with alkyl and OH Selected from; X is O or -CH2-; W is the ring system A: [ka] and; R 4 is cyano; R 5 is H; R 6 is OH); and pharmaceutically acceptable salts thereof.

[0055] Specific examples of compounds of formula I described herein include: 4-[8-[(3R)-1-Ethyl-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; Formic acid; 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 3-Hydroxy-4-[8-(3-hydroxy-3-methyl-cyclobutyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; and pharmaceutically acceptable salts thereof.

[0056] Other specific examples of compounds of formula I described herein are: 3-Hydroxy-4-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile;2,2,2-Trifluoroacetic acid; 3-Hydroxy-4-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol;2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol;2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol; 4-[8-[(3R,5S)-1-ethyl-5-hydroxy-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethyl)phenol; 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid; 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol; 2,2,2-trifluoroacetic acid 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol;2,2,2-trifluoroacetic acid; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol; 4-[8-[(3R)-1-Ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-benzonitrile;2,2,2-Trifluoroacetic acid; 4-[8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-benzonitrile; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol;2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol; 3-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol;2,2,2-trifluoroacetic acid; 3-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol; 5-[8-[(1R,3S)-3-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; 5-[8-[(3S,4R)-3-hydroxytetrahydropyran-4-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; and pharmaceutically acceptable salts thereof.

[0057] Other specific examples of compounds of formula I described herein are: 5-[8-(4-hydroxycyclohexyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; and pharmaceutically acceptable salts thereof.

[0058] Preferred examples of compounds of formula I described herein are: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol;2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol; and pharmaceutically acceptable salts thereof.

[0059] 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 compound 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 dosage and concentration used. The pH of the formulation will depend primarily on the particular application and the concentration of the compound, but preferably ranges from about 3 to about 8. In one example, the compound of Formula I is formulated in an acetate buffer at pH 5. In another embodiment, the compound of Formula I is sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0060] The compositions are formulated, dosed, and administered in a manner consistent with good medical practice. Factors to consider in this context 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 scheduling, and other factors known to medical practitioners.

[0061] The compounds of the present invention may 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 desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0062] The compounds of the present invention may be administered in any convenient dosage form, such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain ingredients conventional in pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and additional active agents.

[0063] 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, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifying agents, glidants, processing aids, colorants, sweeteners, flavors, flavorings, diluents, and other known additives to present the drug (i.e., the compound of the present invention or a pharmaceutical composition thereof) aesthetically or to aid in the manufacture of the pharmaceutical product (i.e., a drug product).

[0064] 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, dragees, hard gelatin capsules, injectable solutions or topical preparations, such as lactose, corn starch or its derivatives, talc, stearic acid or its salts, and can be used, for example, as such adjuvants for tablets, dragees and hard gelatin capsules.

[0065] Suitable adjuvants for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semisolid and liquid polyols, etc.

[0066] Suitable adjuvants for the production of solutions and syrups are, for example, water, polyols, saccharose, invert sugar, glucose etc.

[0067] Suitable adjuvants for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils and the like.

[0068] Suitable adjuvants for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solid or liquid polyols etc.

[0069] Suitable adjuvants for topical ophthalmic formulations are, for example, cyclodextrin, mannitol, or many other carriers and excipients known in the art.

[0070] 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.

[0071] The dosage can vary widely and will, 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 equal amounts. For topical administration, the formulation may contain 0.001% to 15% by weight of the pharmaceutical agent, and the required dose, which may be between 0.1 and 25 mg, may be administered either in a single dose per day or per week, or in multiple doses (2 to 4 times) per day or per week. However, it will be apparent that the upper or lower limits given herein may be exceeded if indicated.

[0072] An embodiment of the present invention is a compound according to Formula I as described herein for use as a therapeutically active substance.

[0073] An 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.

[0074] An 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.

[0075] 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.

[0076] 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 (Menu et al., Clinical and Experimental Immunology, 166:1-15, 2011; Strowig et al., Nature, 481:278-286, 2012).

[0077] In one embodiment, the disease, disorder or condition is selected from the following: (i) Inflammation; (ii) autoimmune diseases; (iii) cancer; (iv) infectious diseases; (v) central nervous system disorders; (vi) metabolic diseases; (vii) cardiovascular disease; (viii) respiratory diseases; (ix) liver disease; (x) Kidney disease; (xi) eye diseases; (xii) skin diseases; (xiii) lymphatic status; (xiv) mental disorder; (xv) graft-versus-host disease; (xvi) allodynia; (xvii) diabetes-related conditions; and (xviii) Any disease in which an individual is determined to have a germline or somatic non-silent mutation in NLRP3.

[0078] In another embodiment, the disease, disorder or condition is selected from the following: (i) Cancer; (ii) infectious diseases; (iii) central nervous system disorders; (iv) cardiovascular disease; (v) liver disease; (vi) eye disease; and (vii) Skin diseases.

[0079] 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) neurological conditions such as multiple sclerosis or encephalomyelitis; (x) 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 co-infection), 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; (xi) 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; (xii) lymphatic conditions such as Castleman's disease; (xiii) a condition of the immune system, or a condition involving it, such as hyper-IgE syndrome, lepromatous leprosy, familial hemophagocytic lymphohistiocytosis, or graft-versus-host disease; (xiv) 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; (xv) Cancer, including those listed above; (xvi) burns, wounds, trauma, hemorrhage, or stroke; (xvii) radiation exposure; (xviii) metabolic diseases such as type 2 diabetes (T2D), atherosclerosis, obesity, gout, or pseudogout; and / or (xix) Pain such as inflammatory hyperalgesia, pelvic pain, allodynia, neuropathic pain, or cancer-induced bone pain.

[0080] An 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 the following: inflammation; autoimmune diseases; cancer; infectious diseases; central nervous system diseases; metabolic diseases; cardiovascular disease; respiratory diseases; liver disease; kidney disease; eye diseases; skin diseases; Lymphatic condition; Mental disorders; graft-versus-host disease; allodynia; diabetes-related conditions; and Any disease in which an individual is determined to have a germline or somatic non-silent mutation in NLRP3.

[0081] An embodiment of the present invention is the use of a compound according to formula I as described herein in the treatment or prevention of a disease, disorder or condition, which disease, disorder or condition is responsive to NLRP3 inhibition.

[0082] An embodiment of the present invention is the use of a compound according to Formula I as described herein in the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

[0083] An 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.

[0084] An 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 Alzheimer's disease and Parkinson's disease.

[0085] An 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.

[0086] An 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 Alzheimer's disease and Parkinson's disease.

[0087] An 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] An embodiment of the present invention is a method for the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease, the method comprising administering an effective amount of a compound according to Formula I as described herein.

[0089] An embodiment of the present invention is a method of treating or preventing a disease, disorder or condition selected from asthma and COPD, the method comprising administering an effective amount of a compound according to formula I as described herein.

[0090] An embodiment of the present invention relates to a method of inhibiting NLRP3, the method comprising administering an effective amount of a compound according to Formula I described herein.

[0091] Also an embodiment of the invention is a compound of formula I as described herein when prepared according to any one of the processes described.

[0092] An embodiment of the present invention is a pharmaceutical composition comprising a compound according to Formula I, as described herein, and a therapeutically inert carrier.

[0093] Assay procedure NLRP3 and pyroptosis It is well established that activation of NLRP3 leads to cell pyroptosis, a hallmark of which 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, it is expected that inhibitors of NLRP3 will block pyroptosis and the release of pro-inflammatory cytokines (e.g., IL-1β) from cells.

[0094] 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 THP-1 cells were grown to a concentration of 625,000 cells / ml. On the day of the experiment, the cells were harvested and resuspended in RPMI medium (without FBS). The 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 plates prepared in this manner were used for compound screening.

[0095] THP-1 cell pyroptosis assay For compound screening, the following method stepwise assay was followed. THP-1 cells (25,000 cells / well) are seeded in 40 μl of RPMI medium (without FBS) containing 1.0 μg / ml LPS in 96-well black-walled, clear-bottom cell culture plates coated with poly-D-lysine (VWR no. 734-0317). 5 μl of compound (8-point half-log dilutions with the highest dose at 10 μM) or vehicle (FAC in DMSO 0.1%) 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)

[0096] The results of the pyroptosis assay were compared with those of THP IC 50 These are summarized in Table 1 below.

[0097] 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 several compounds in human whole blood according to the following protocol.

[0098] Human whole blood in Li-heparin tubes was obtained from healthy donors from a volunteer donor panel. Plate out 80 μl of whole blood containing 1 μg / ml LPS into a 96-well clear-bottom cell culture plate (Corning #3585). 10 μl of compound (8-point half-log dilutions at the highest dose of 10 μM) or vehicle (FAC in DMSO 0.1%) is added to the appropriate wells. Incubate at 37°C, 5% CO2 for 3 hours. Add 10 μl of nigericin (Sigma #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 minutes to pellet the cells, and 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)

[0099] Human whole blood assay results HWB IC 50 These are summarized in Table 1 below.

[0100] hERG screening assay Cardiac arrhythmias are one of the most common side effects leading to drug failure during small molecule drug development. 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.

[0101] 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.

[0102] 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.

[0103] electrophysiology The effects of compounds on hERG K+-current parameters will be evaluated at two concentrations in at least four cells.

[0104] 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.

[0105] 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.

[0106] 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]

[0107] 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).

[0108] The results of the hERG assay were analyzed using the hERG IC 20 The results are summarized in Table 2 below.

[0109] Intracellular 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.

[0110] P-gp is expressed in the apical membrane of the monolayer.

[0111] The adhesion of the cell monolayer and the functional activity of P-gp are confirmed by the addition of the cell-impermeable marker Lucifer Yellow and the reference P-gp substrate edoxaban, respectively.

[0112] 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 intracellular 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.

[0113] 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.

[0114] Microsomal stability: Incubations of test compounds at 1 μM 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 compounds, 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. Samples are then cooled and centrifuged, after which the supernatants are analyzed by LC-MS / MS2.

[0115] Metabolic stability in hepatocytes: Assay Description: Biological materials. Cryopreserved hepatocytes [mouse, rat, rabbit, monkey, human (mixed sex)] are obtained. Hepatocyte viability after reconstitution is at least 80% throughout the study. Ready-to-use rat / human HepatoPac® cultures [long-term hepatocyte co-cultures; pooled (for humans, n=5 males and n=5 females)] containing stromal mouse fibroblasts (negative control; pooled) are obtained, along with incubation plates, application medium, and maintenance medium.

[0116] 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; 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 compounds (e.g., 1 μM) to wells (1 × 105 cells / well), the 96-well hepatocyte suspension culture plate was incubated at 37 °C in 5% CO2. Samples were quenched at designated time points up to 2 hours by adding acetonitrile (containing an internal standard) to the incubation wells.

[0117] Incubation of test substances (e.g., 1 μM, 0.1% v / v DMSO) performed in metabolic suspension assays using HepatoPac® is carried out in 96-well plates containing either co-cultures of adherent hepatocytes and mouse fibroblast control cells, or control cells alone (5% CO2 atmosphere and 37°C). The incubation medium for human HepatoPac® is the same as 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.

[0118] The samples are then appropriately centrifuged and the supernatants are analyzed by LC-MS / MS. n=1 or 2 incubations are performed. [Table 2] [Table 3] [Brief explanation of the drawings]

[0119] [Figure 1] Figure 1 shows the voltage pattern used for cells that were held at a resting voltage of -80 mV (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. DETAILED DESCRIPTION OF THE INVENTION

[0120] The invention will now be illustrated by the following examples, which have no limiting character.

[0121] Where preparations are obtained as mixtures of enantiomers or diastereomers, the pure enantiomers or diastereomers may be obtained by the methods described herein or by methods known to those skilled in the art, such as, for example, chiral chromatography or crystallization.

[0122] Experimental Method [Table 4] Unless otherwise specified, all examples and intermediates were prepared under a nitrogen atmosphere.

[0123] Intermediates: Intermediate 1: (4-cyano-2-hydroxy-6-methyl-phenyl)boronic acid [ka] Step A: 4-amino-3-methoxy-5-methyl-benzonitrile Two batches were run in parallel.

[0124] To a solution of commercially available 4-bromo-2-methoxy-6-methylbenzenamine (CAS No. 348169-39-1, 25.0 g, 115 mmol, 1.00 equiv.) in DMF (250 mL) was added Zn(CN) (13.5 g, 115 mmol, 7.34 mL, 1.00 equiv.) and Pd(PPh) (66.8 g, 57.8 mmol, 0.50 equiv.). The reaction mixture was stirred at 100 °C for 12 h. The reaction mixture was poured into water (1.50 L) and extracted with ethyl acetate (1 L × 3). The organic phase was washed with brine (1 L × 3), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue that was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 100 / 1 to 0 / 1) to give the title compound (28.0 g, 75% yield) as a yellow solid. 1 H NMR(DMSO-d6)δ 7.05(s,2H),5.47(bs,2H),3.81(s,3H),2.09(s,3H).

[0125] Step B: 4-Bromo-3-methoxy-5-methyl-benzonitrile To a solution of CuBr (46.4 g, 323 mmol, 9.86 mL, 1.50 equiv) in MeCN (180 mL), t-BuONO (33.3 g, 323 mmol, 38.5 mL, 1.50 equiv) was added and stirred at 65 °C. Then, a solution of the above intermediate 2B 4-bromo-3-methoxy-5-methyl-benzonitrile (35.0 g, 215 mmol, 1.00 equiv) in MeCN (180 mL) was added at 65 °C. The mixture was stirred at 65 °C for 3.5 h. After completion, saturated aqueous NaSO (400 mL) and saturated aqueous NH Cl (200 mL) were added to the mixture, which was then extracted with ethyl acetate (500 mL × 3). The organic phase was washed with brine (500 mL × 2), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=100 / 1 to 0 / 1, Rf=0.75) to give the title compound (20.7 g, yield 42%) as a white solid. 1 H NMR(DMSO-d6)δ 7.43,7.40(2s,1H each),3.90(s,3H),2.37(s,3H).

[0126] Step C: 3-Methoxy-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile To a solution of the above 4-bromo-3-methoxy-5-methyl-benzonitrile (18.0 g, 79.6 mmol, 1.00 equiv.) in DMF (180 mL) was added B2Pin2 (30.3 g, 119 mmol, 1.50 equiv.) and AcOK (35.1 g, 358 mmol, 4.50 equiv.). The mixture was stirred at 20 °C for 0.5 h, and Pd(dppf)Cl2·CHCl2 (13.0 g, 15.9 mmol, 0.20 equiv.) was added. The mixture was stirred at 100 °C for 12 h. The mixture was filtered through diatomaceous earth, diluted with HO (500 mL), and extracted with ethyl acetate (800 mL × 3). The organic phase was washed with brine (800 mL × 3), dried over NaSO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate=100 / 1 to 1 / 1, Rf=0.30) to give the title compound (18.0 g, yield 83%) as a white solid. 1 H NMR(DMSO-d6)δ 7.22,7.21(2s,1H each),3.75(s,3H),2.27(s,3H),1.30(s,12H).

[0127] Step D: (4-cyano-2-hydroxy-6-methyl-phenyl)boronic acid A solution of the above 3-methoxy-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (17.0 g, 96.0 mmol, 1.00 equiv) in dichloromethane (170 mL) was cooled to 0 °C, and BBr (38.9 g, 155 mmol, 2.50 equiv) was added dropwise at 0 °C. The mixture was stirred at 0 °C for 0.5 h. The mixture was poured into HO (200 mL), filtered, and the cake collected and triturated with EtOAc (20 mL) to give the title compound (4.67 g, 42% yield) as a gray solid. LCMS: m / z 178.1 [M+H] + ,ESI pos.

[0128] Intermediate 2: 3,6-dichloro-4-(3-chloropropyl)pyridazine [ka] To a stirred suspension of 3,6-dichloropyridazine (CAS No. 141-30-0, 5.0 g, 33.6 mmol, 1.0 equiv.) in water (125 mL) was added H2SO4 (6.6 g, 67.1 mmol, 2.0 equiv.). After heating the mixture to 70 °C, 4-chlorobutyric acid (CAS No. 627-00-9, 4.50 g, 36.9 mmol, 1.1 equiv.) was added, followed by a solution of silver nitrate (1.30 g, 7.65 mmol, 0.23 equiv.) in water (2.5 mL) over 1 min. At this point, the mixture became milky white in appearance, and a solution of ammonium persulfate (25.8 g, 113.1 mmol, 3.40 equiv.) in water (65 mL) was added slowly over 20–30 min, causing the product to form a sticky precipitate. The reaction mixture was stirred at 70 °C for 1 h. After the reaction was complete, the mixture was cooled to room temperature, poured into ice, and basified with concentrated aqueous ammonia to adjust the pH to approximately 8, maintaining the temperature below 5° C. The aqueous phase was extracted with dichloromethane (3×400 mL), and the combined extracts were dried over NaSO, filtered, concentrated under reduced pressure, and purified by reverse-phase flash (CombiFlash 0.1% TFA aqueous-ACN conditions), followed by lyophilization to give the title compound (4.20 g, 56% yield) as a colorless oil. LCMS: m / z 226.9 [M+H] + ,ESI pos.

[0129] Intermediate 3: 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((2-(trimethylsilyl)ethoxy)methoxy)benzonitrile [ka] Step A: 4-Bromo-2-methoxy-6-methylaniline To a solution of 2-methoxy-6-methylaniline (CAS number 50868-73-0, 50.0 g, 364.5 mmol, 1.0 equiv.) in methanol (150 mL) and acetic acid (50.0 mL, 874.2 mmol, 2.4 equiv.), Br2 (22.4 mL, 437.4 mmol, 1.2 equiv.) was slowly added dropwise at 0 °C, and then stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was diluted with water (300 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (hexane / EtOAc, 1:0 to 10:1), then slurried with petroleum ether (30 mL), filtered, and the cake was collected to give the title compound (30.3 g, 39% yield) as a brown solid. 1 H NMR(400MHz,DMSO-d6)δ 6.82(d,1H),6.78(d,1H),4.57(m,2H),3.77(s,3H),2.06(s,3H).

[0130] Step B: 4-amino-3-methoxy-5-methylbenzonitrile To a solution of the aforementioned 4-bromo-2-methoxy-6-methyl-aniline (30.3 g, 140.2 mmol, 1.0 equiv.) in DMF (260 mL) was added Zn(CN) (16.5 g, 140.5 mmol, 1.00 equiv.) and Pd(PPh) (32.4 g, 28.1 mmol, 0.2 equiv.). The mixture was stirred at 100 °C under a nitrogen atmosphere for 12 h. After the reaction was complete, the mixture was cooled to 20 °C, poured into HO (500 mL), and extracted with ethyl acetate (300 mL × 3). The organic phase was washed with brine (200 mL × 3), dried over NaSO, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (hexane / EtOAc, 1:0 to 5:1) to give the title compound (22.0 g, 97% yield) as a pink solid. 1 H NMR(400MHz,DMSO-d6)δ 7.05(s,2H),5.48(s,2H),3.81(s,3H),2.09(s,3H).

[0131] Step C: 4-Bromo-3-methoxy-5-methylbenzonitrile To a solution of the above 4-amino-3-methoxy-5-methyl-benzonitrile (22.0 g, 135.6 mmol, 1.0 equiv.) and CuBr (29.2 g, 203.5 mmol, 1.5 equiv.) in ACN (220 mL), butyl nitrite (20.98 g, 203.47 mmol, 1.5 equiv.) was added, and the mixture was stirred at 65 °C for 2 h. After the reaction was complete, the mixture was cooled to 20 °C, diluted with ACN (150 mL), filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (hexane / EtOAc, 1:0 to 5:1). The residue was then slurried with a mixture of ethyl acetate (30 mL) and petroleum ether (3 mL), filtered, and the cake was collected to give the title compound (18.0 g, 59% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ 7.46(s,1H),7.43(s,1H),3.91(s,3H),2.39(s,3H).

[0132] Step D: 4-Bromo-3-hydroxy-5-methylbenzonitrile To a solution of the above 4-bromo-3-methoxy-5-methyl-benzonitrile (6.0 g, 26.5 mmol, 1.0 equiv.) in DCM (30 mL) was added BBr3 (30.0 mL, 316.1 mmol, 11.9 equiv.) at 0 °C, and the mixture was stirred at 25 °C for 1 h. After the reaction was completed, it was combined with another batch (10 g), then diluted with water (500 mL), and extracted with ethyl acetate (150 mL × 3). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (hexane / EtOAc, 1:0 to 10:1) to give the title compound (13.3 g, 89% yield) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ 10.15(s,1H),6.42(d,1H),6.24(d,1H),1.51(s,3H).

[0133] Step E: 4-Bromo-3-methyl-5-((2-(trimethylsilyl)ethoxy)methoxy)benzonitrile To a solution of the above 4-bromo-3-hydroxy-5-methyl-benzonitrile (13.3 g, 62.7 mmol, 1.0 equiv.) in DMF (60 mL), CsCO (33.3 g, 102.1 mmol, 1.63 equiv.) was added at 0° C., followed by stirring at 20° C. for 0.5 h, and then 2-(trimethylsilyl)ethoxymethyl chloride (15.5 mL, 87.8 mmol, 1.4 equiv.) was added dropwise at 0° C., followed by stirring at 20° C. for 1 h. After the reaction was complete, it was diluted with water (200 mL) and extracted with MTBE (150 mL×3). The combined organic phase was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (hexane / EtOAc, 1:0 to 10:1) to give the title compound (16.3 g, 76% yield) as a white solid. 1 H NMR(400MHz,CDCl3)δ 7.27(d,1H),7.18(d,1H),5.30(s,2H),3.77(t,2H),2.44(s,3H),0.95(t,2H),0(s,9H).

[0134] Step F: 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((2-(trimethylsilyl)ethoxy)methoxy)benzonitrile To a solution of the aforementioned 4-bromo-3-methyl-5-(2-trimethylsilylethoxymethoxy)benzonitrile (10.0 g, 29.2 mmol, 1.0 equiv.) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (14.8 g, 58.4 mmol, 2.0 equiv.) in 1,4-dioxane (100 mL) was added CsCO (19.0 g, 58.4 mmol, 2.0 equiv.) and tris(4-methoxy-3,5-dimethylphenyl)phosphane (CAS No. 121898-64-4, 1.28 g, 2.92 mmol, 0.1 equiv.). Pd(OAc) (0.66 g, 2.92 mmol, 0.1 equiv) was added under N2 and stirred at 95 °C for 5 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water (150 mL) and ethyl acetate (50 mL), filtered, and the filtrate was extracted with ethyl acetate (100 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 and purified by column chromatography (hexane / EtOAc, 1:0 to 10:1) (17 g), then triturated with MeOH (51 mL), filtered, and the cake was collected to give the title compound (8.38 g, 74% yield) as a white solid. 1 H NMR(400MHz,CDCl3)δ 7.13(s,1H),7.07(s,1H),5.18(s,2H),3.72(t,2H),2.36(s,3H),1.38(s,12H),0.94(t,2H),0.00(s,9H).

[0135] Intermediate 4: (1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexanamine [ka] To a solution of 2-aminocyclohexanol (10.0 g, 86.83 mmol, 1.0 equiv.) in DCM (100 mL), TEA (14.5 mL, 104.2 mmol, 1.2 equiv.) and tert-butyldimethylchlorosilane (15.7 g, 104.2 mmol, 1.2 equiv.) were added, and the mixture was stirred at 20 °C for 12 h. After the reaction was completed, the reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (60 mL × 3). The combined organic phase was washed with brine (100 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified using a silica gel column (petroleum ether:ethyl acetate = 1:0 to 3:1) to give the title compound (20.5 g, 82% yield) as a yellow solid. 1 H NMR(400MHz,CDCl3)δ[ppm]:3.20-3.14(m,1H),2.54-2.48(m,1H),1.86-1.82(m,2H),1 .70-1.50(m,2H),1.28-1.21(m,3H),1.14-1.04(m,1H),0.90(s,9H),0.09-0.07(d,6H).

[0136] Intermediate 5: tert-Butyl-[(1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane [ka] Step A: Methyl 2-(3,6-dichloropyridazin-4-yl)oxyacetate To a solution of methyl glycolate (CAS no. 96-35-5, 16.4 mL, 212.6 mmol, 1.0 equiv.) in THF (100 mL), NaH (12.8 g, 318.9 mmol, 1.5 equiv., purity: 60% in mineral oil) was added portionwise at 25 °C under N , and the resulting mixture was stirred at 25 °C for 10 min. Then, 3,4,6-trichloropyridazine (CAS no. 6082-66-2, 39.0 g, 212.6 mmol, 1.0 equiv.) was added to the above mixture, and the reaction mixture was stirred at 25 °C under N for 1 h. After the reaction was complete, the mixture was quenched with HO (100 mL) and extracted with EtOAc (150 mL × 3). The combined organic phase was washed with brine (100 mL × 2), dried over anhydrous Na SO , filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1 to 2:1) to give the title compound (20.3 g, 40% yield) as a yellow solid. LCMS: 236.9 [M+H] + ,ESI pos.

[0137] Step B: 2-(3,6-dichloropyridazin-4-yl)oxyacetaldehyde A solution of methyl 2-(3,6-dichloropyridazin-4-yl)oxyacetate (0.18 g, 0.76 mmol, 1 equiv.) in THF (4 mL) was cooled to −65° C., and then DIBAL-H (1 M in toluene, 2.1 mL, 2.1 mmol, 2.77 equiv.) was added dropwise to the above mixture under N. The reaction mixture was then stirred at −65° C. for 2 hours under N. After the reaction was completed, the reaction mixture was quenched with ice water (5 mL) at 0° C. and extracted with EtOAc (15 mL×2). The combined phases were washed with brine (10 mL×2), filtered, and the filtrate was dried over NaSO, filtered, and concentrated in vacuo to give the crude product (180 mg, 57% yield) as a yellow oil. The crude product was used directly in the next step without further purification.

[0138] Step C: (1R,2R)-2-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]-cyclohexanamine To a solution of 2-(3,6-dichloropyridazin-4-yl)oxyacetaldehyde (180 mg, crude from the above step) in DCE (4 mL) was added trans-(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexanamine (199.5 mg, 0.87 mmol, 1.0 equiv.). The mixture was stirred at 20° C. for 15 hours, and then NaBH(OAc) (460.7 mg, 2.17 mmol, 2.5 equiv.) was added to the above mixture. The reaction mixture was stirred at 20° C. for 15 minutes. After the reaction was complete, the reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL × 2). The combined extracts were washed with brine (10 mL), filtered, and the filtrate was dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate=1:1) to give the title compound (50.0 mg, 12% yield over two steps) as a yellow oil. LCMS: m / z 420.2 [M+H] + ,ESI pos.

[0139] Step D: tert-Butyl-[(1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane To a solution of (1R,2R)-2-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]-cyclohexanamine (100.0 mg, 0.24 mmol, 1.0 equiv.), CsCO (147.2 mg, 0.45 mmol, 1.9 equiv.) in 1,4-dioxane (2 mL) was added Pd-PEPPSI-IHEPTCl (19.9 mg, 0.02 mmol, 0.1 equiv.) under N. The reaction mixture was then stirred at 100 °C under N for 4 h. After the reaction was complete, the reaction mixture was cooled to room temperature, poured into ice (20 mL), and extracted with ethyl acetate (50 mL × 2). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate=5:1, Rf=0.6) to give the title compound (15.0 mg, 12% yield) as a yellow oil. LCMS: m / z 384.2, [M+H] + ,ESI pos.

[0140] Intermediate 6: tert-butyl-[(1R,2R)-2-(3-chloro-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane [ka] Step A: 2-(3,6-dichloropyridazin-4-yl)oxyethanol To a solution of methyl 2-(3,6-dichloropyridazin-4-yl)oxyacetate (16.5 g, 69.6 mmol, 1.0 equiv.) in methanol (180 mL) and THF (90 mL) was added NaBH4 (5266.6 mL, 139.2 mmol, 2.0 equiv.) at 0 °C, and the reaction mixture was then stirred at 25 °C for 1 h. After the reaction was complete, the mixture was quenched with saturated aqueous NH4Cl (100 mL) and washed with EtOAc (100 mL × 2). The organic phase was washed with brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1 to 1:1) to give the title compound (9.8 g, 67% yield) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ[ppm]:7.75(s,1H),5.03(t,1H),4.32(t,2H),3.80-3.70(m,2H).

[0141] Step B: 2-(3,6-dichloropyridazin-4-yl)oxyethyl ethanesulfonate To a solution of 2-(3,6-dichloropyridazin-4-yl)oxyethanol (1.0 g, 4.78 mmol, 1.0 equiv.) in DCM (10 mL) was added DIEA (2.37 mL, 14.35 mmol, 3.0 equiv.), and then a solution of ethanesulfonyl chloride (CAS No. 594-44-5, 0.68 mL, 7.18 mmol, 1.5 equiv.) in DCM (2 mL) was added dropwise, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was complete, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phase was washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate 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 (1.0 g, 63% yield) as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ[ppm]:7.80(s,1H),4.65-4.52(m,4H),3.44-3.37(m,2H),1.26(t,3H).

[0142] Step C: 2-(3,6-dichloro-5-methyl-pyridazin-4-yl)oxyethyl ethanesulfonate To a stirred solution of 2-(3,6-dichloropyridazin-4-yl)oxyethyl ethanesulfonate (4.0 g, 13.3 mmol, 1.0 equiv.) and silver nitrate (1.53 g, 9.01 mmol, 0.68 equiv.) in water (30 mL) was added CH3COOH (2.28 mL, 39.9 mmol, 3.0 equiv.). The mixture was heated to 50°C, and then a solution of sulfuric acid (2.17 mL, 39.9 mmol, 3.0 equiv.) in water (15 mL) was added dropwise to the mixture. The temperature was then raised to 70°C, and a solution of ammonium persulfate (9093.1 mg, 39.85 mmol, 3.0 equiv.) in water (15 mL) was added dropwise to the mixture over 30 minutes. The reaction mixture was then heated to 70°C and stirred for an additional 30 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. EtOAc (150 mL) was added to the mixture, and the precipitate was filtered off. The filtrate was washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1) to give the title compound (1200.0 mg, 26% yield) as a colorless oil. LCMS: m / z 315.0 [M+H] + ,ESI pos.

[0143] Step D: tert-Butyl-[(1R,2R)-2-(3-chloro-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane To a solution of 2-(3,6-dichloro-5-methyl-pyridazin-4-yl)oxyethyl ethanesulfonate (1000.0 mg, 3.17 mmol, 1.0 equiv.), trans-(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexanamine (1000.0 mg, 4.36 mmol, 1.37 equiv.) in DMF (5 mL) was added DIEA (736.8 mg, 5.71 mmol, 1.8 equiv.), and the reaction mixture was then stirred at 80° C. under N for 12 h. After the reaction was complete, the reaction mixture was subjected to reverse-phase flash chromatography (Xtimate C column). 18 , 250 mm × 50 mm × 10 μm; mobile phase: [water (0.1% ammonia hydroxide, v / v)-ACN]; B%: 5% to 70%, 10 min) to give the title compound (15.0 mg, 1% yield) as a white solid. LCMS: m / z 398.2 [M+H] + ,ESI pos.

[0144] Intermediate 7: (3S,4R)-3-[tert-butyl(dimethyl)silyl]oxytetrahydropyran-4-amine [ka] To a solution of trans-4-aminocyclohexanol (3.0 g, 26.05 mmol, 1.0 equiv.) in DCM (30 mL) was added TEA (4.36 mL, 31.26 mmol, 1.2 equiv.) and tert-butyldimethylchlorosilane (4.71 g, 31.26 mmol, 1.2 equiv.), followed by stirring at 20 °C for 12 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column (ethyl acetate:MeOH = 1:0 to 0:1) to give the title compound (2.1 g, 32% yield) as a yellow oil.

[0145] Intermediate 8: (1R,3S)-3-[tert-butyl(dimethyl)silyl]oxycyclohexanamine [ka] To a solution of (1S,3R)-3-aminocyclohexanol (0.9 g, 7.81 mmol, 1.0 equiv) in DCM (10 mL) was added TEA (3.27 mL, 23.44 mmol, 3.0 equiv) and tert-butyldimethylchlorosilane (1.41 g, 9.38 mmol, 1.2 equiv), and the reaction was stirred at 20 °C for 12 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was combined with another batch, and the combined crude was purified by silica gel column (PE:EA = 1:0 to 0:1) to give the title compound as a colorless oil (1.2 g, 60% yield).

[0146] Intermediate 9: trans-4-[tert-butyl(dimethyl)silyl]oxycyclohexanamine [ka] To a solution of trans-4-aminocyclohexanol (3.0 g, 26.05 mmol, 1.0 equiv.) in DCM (30 mL), TEA (4.36 mL, 31.26 mmol, 1.2 equiv.) and tert-butyldimethylchlorosilane (4.71 g, 31.26 mmol, 1.2 equiv.) were added, and the mixture was stirred at 20 °C for 12 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate:MeOH = 1:0 to 0:1) to give the title compound as a yellow solid (2.1 g, 32% yield). [Example]

[0147] Example 1: 4-[8-[(3R)-1-Ethyl-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; Formic acid [ka] Step A: (3S)-3-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]piperidine-1-carboxylic acid tert-butyl ester According to WO 2020190793, to a solution of 3,6-dichloro-4-(2-iodoethoxy)pyridazine (CAS No. 2490352-76-4, prepared according to WO 2020190793, 300 mg, 893.6 umol, 1.00 eq) and (3R)-3-aminopiperidine-1-carboxylic acid tert-butyl ester (CAS No. 188111-79-7, 196.7 mg, 189.3 uL, 982.9 umol, 1.10 eq) in DMSO (5 mL) was added N,N-diisopropylethylamine (312 uL, 1.79 mmol, 2.00 eq). The mixture was stirred at 50° C. for 4 hours and then filtered. The reaction was quenched with water (50 mL) and extracted with ethyl acetate (2 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel flash chromatography using gradient elution (solid deposit, 30-80% ethyl acetate / heptane) to afford the title compound (202 mg, 58%) as a yellow oil. LCMS: m / z 391.3 [M+H] + ,ESI pos.

[0148] Step B: (3R)-3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)piperidine-1-carboxylic acid tert-butyl ester According to WO 2020190793, a mixture of the aforementioned (3S)-3-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]piperidine-1-carboxylic acid tert-butyl ester (202 mg, 516.2 μmol, 1.00 equiv.), palladium(II) acetate (CAS No. 3375-31-3, 9.27 mg, 41.3 μmol, 0.08 equiv.), (R)-(+)-Binap (CAS No. 76189-55-4, 41.79 mg, 67.1 μmol, 0.130 equiv.), and cesium carbonate (319.6 mg, 980.9 μmol, 1.90 equiv.) in toluene (4 mL) was degassed three times with N and stirred at 110 °C for 3 h. After cooling to room temperature, the reaction mixture was diluted with water and extracted with ethyl acetate (2 × 20 mL). The crude material was purified by column flash chromatography on silica gel (24 g, 0-50% EtOAc in Heptane) to give the title compound (54 mg, 29% yield) as a white powder. LCMS: m / z 355.3 [M+H] + ,ESI pos.

[0149] Step C: (3R)-3-[3-(4-cyano-2-hydroxy-6-methyl-phenyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]piperidine-1-carboxylic acid tert-butyl ester A mixture of the aforementioned (3R)-3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)piperidine-1-carboxylic acid tert-butyl ester (54 mg, 0.152 mmol, 1.00 equiv.), (4-cyano-2-hydroxy-6-methyl-phenyl)boronic acid intermediate 1 (45.8 mg, 258.7 μmol, 1.70 equiv.), and cesium carbonate (148.8 mg, 456.6 μmol, 3.00 equiv.) in 1,4-dioxane (2 mL) and water (400 μL) was stirred in a sealed tube. Argon was bubbled through the mixture for 2 minutes, and then catalyst (XPhos Pd G3) (19.3 mg, 22.8 μmol, 0.15 equiv.) was finally added. The sealed tube was stirred at 100° C. for 3 hours. The reaction mixture was extracted with ethyl acetate (2 x 80 mL) and semi-saturated NH4Cl solution (20 mL). The organic layer was washed with water (20 mL) and brine (20 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel flash chromatography using gradient elution (solid deposit, 0-50% ethyl acetate / heptane) to afford the title compound (17 mg, 25%) as a pale yellow powder. LCMS: m / z 452.4 [M+H] + ,ESI pos.

[0150] Step D: 3-Hydroxy-5-methyl-4-[8-[(3R)-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]benzonitrile 1:1 Hydrochloric acid To a solution of the aforementioned (3R)-3-[3-(4-cyano-2-hydroxy-6-methyl-phenyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]piperidine-1-carboxylic acid tert-butyl ester (17 mg, 37.7 μmol, 1.00 equiv.) in dichloromethane (0.2 mL) and methanol (0.1 mL), hydrochloric acid (4 M in dioxane, 75.3 μL, 301.2 μmol, 8.00 equiv.) was added dropwise at room temperature. The reaction mixture was stirred at 23 °C for 2 hours. The mixture was concentrated in vacuo to give the crude product (150 mg) as a yellow oil, which was used in the next step without further purification. LCMS: m / z 348.3 ([{35Cl}MH]-), 350.3 ([{37Cl}MH]-), ESI neg.

[0151] Step E: 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; formic acid To a suspension of the aforementioned 3-hydroxy-5-methyl-4-[8-[(3R)-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]benzonitrile hydrochloride (18 mg, 46.4 μmol, 1.00 equiv.) in dichloromethane (0.1 mL) was added triethylamine (9.65 μL, 69.6 μmol, 1.50 equiv.). Then, under ice cooling, acetaldehyde (6.51 μL, 116.02 μmol, 2.50 equiv.) was added, followed by sodium triacetoxyborohydride (17.7 mg, 83.5 μmol, 1.80 equiv.). The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 45 minutes. After completion of the reaction, the reaction mixture was extracted with DCM, ammonium chloride, and water. The aqueous layer was back-extracted twice with DCM. The combined organic layers were washed with water and brine, then dried over sodium sulfate, filtered, concentrated in vacuo, and finally purified using RP HPLC (Gemini NX, 12 nm, 5 μm, 100×30 mm; ACN / water + 0.1% HCOOH with a gradient of 5-50 CAN) to give the title compound as a 1:1 formic acid (9 mg, 41%) as a light brown powder. LCMS: m / z 378.3 [M+H] + ,ESI pos. 1 H NMR(600MHz,DMSO-d6)δ ppm 10.31(br s,1H),9.56-9.85(m,1H),7.19-7.27(m,1H),7.12(d,1H),6.82-6.89(m,1H),5.15-5.19(m,1H),4.23-4.42(m,2H),3.52- 3.63(m,2H),3.40-3.52(m,1H),3.03-3.24(m,2H),2.70-2.97(m,1H),2.10(s,3H),1.60-2.06(m,5H),1.23-1.25(m,2H).

[0152] Example 2: 4-[8-[(3R)-1-Ethyl-3-piperidyl]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-3-yl]-3-hydroxy-5-methyl-benzonitrile [ka] Step A: (R)-3-chloro-8-(1-ethylpiperidin-3-yl)-5,6,7,8-tetrahydropyrido[2,3-c]pyridazine To a mixture of 3,6-dichloro-4-(3-chloropropyl)pyridazine intermediate 2 (500.0 mg, 2.22 mmol, 1.0 equiv.), (3R)-1-ethylpiperidin-3-amine (611.8 mg, 4.77 mmol, 2.15 equiv.) in DMF (4 mL), TEA (725 mg, 6.09 mmol, 2.75 equiv.) was added and stirred at 150 °C for 2 h. The reaction mixture was cooled to room temperature and purified by reverse-phase flash (CombiFlash 0.1% aqueous NH3HO-ACN) followed by lyophilization to give the title compound (111 mg, 18% yield) as a white solid. LCMS: m / z 281.0 [M+H]+, ESI pos.

[0153] Step B: (R)-4-(8-(1-ethylpiperidin-3-yl)-5,6,7,8-tetrahydropyrido[2,3-c]pyridazin-3-yl)-3-methyl-5-((2-(trimethylsilyl)ethoxy)methoxy)benzonitrile To a solution of the above 3-chloro-8-[(3R)-1-ethyl-3-piperidine]-6,7-dihydro-5H-pyrido[2,3-c]pyridazine (140 mg, 0.50 mmol, 1.0 equiv) and 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(2-trimethylsilylethoxymethoxy)benzonitrile intermediate 3 (291 mg, 0.75 mmol, 1.5 equiv) in 1,4-dioxane (4 mL) and water (0.8 mL) was added CsF (227.2 mg, 1.5 mmol, 3.0 equiv) and Xphos Pd G3 (42.3 mg, 0.05 mmol, 0.1 equiv) and then stirred at 100° C. under nitrogen for 4 hours. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by reverse-phase flash (CombiFlash 0.1% NH3H2O ​​aqueous-ACN condition) and then lyophilized to give the title compound (25.0 mg, 9% yield) as a yellow solid. LCMS: m / z 508.4 [M+H] + ,ESI pos.

[0154] Step C: 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-3-yl]-3-hydroxy-5-methyl-benzonitrile To a solution of the above 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-3-yl]-3-methyl-5-(2-trimethylsilylethoxymethoxy)benzonitrile (20.0 mg, 0.04 mmol, 1.0 equiv.) in DCM (1 mL) was added TFA (0.5 mL) and stirred under nitrogen at 25 °C for 1 h. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give the crude product, which was purified by reverse-phase flash (CombiFlash 0.1% NH3-HO aqueous-ACN conditions) and then lyophilized to give the title compound (13.1 mg, 84% yield) as a yellow solid. LCMS: m / z 378.3 [M+H]+, ESI pos. 1H NMR(400MHz,CD3OD)δ 7.15(s,1H),7.09(s,1H),7.04(s,1H),5.12-5.09(m,1H),3.55-3.49(m,2H),3.11-3.08(m,1H),2.97-2.94(m,1H),2. 81-2.78(m,2H),2.53-2.46(m,2H),2.22-2.20(m,1H),2.17(s,3H),2.00-1.92(m,5H),1.78-1.71(m,2H),1.15(t,3H).

[0155] Example 3: 3-Hydroxy-4-[8-(3-hydroxy-3-methyl-cyclobutyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile [ka] Step A: 3-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]-1-methyl-cyclobutanol Commercially available 3,6-dichloro-4-(2-iodoethoxy)pyridazine (CAS No. 2490352-76-4, 0.54 g, 1.69 mmol, 1.0 equiv.), 3-amino-1-methyl-cyclobutanol (CAS No. 1523606-23-6, 188.4 mg, 1.86 mmol, 1.1 equiv.), and DIPEA (1.18 mL, 6.77 mmol, 4.0 equiv.) were dissolved in DMSO (20 mL) and stirred at 50° C. for 48 h. The mixture was diluted with EtOAc (100 mL), washed with brine (100 mL) and 10 wt % aqueous LiCl (3×50 mL), dried using a phase separator, and concentrated in vacuo. The resulting residue was purified by flash chromatography (silica gel, 24 g, 0-10% MeOH in DCM) to give the title compound (119.0 mg, 23%) as a light brown solid. LCMS: m / z 292.1 ([35Cl][M+H]+, ESI pos.

[0156] Step B: 3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)-1-methyl-cyclobutanol Pd-176 (CAS 879689-47-1, 55.2 mg, 0.07 mmol, 0.2 equiv.), the aforementioned 3-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]-1-methyl-cyclobutanol (100 mg, 0.34 mmol, 1.0 equiv.), and cesium carbonate (334.6 mg, 1.03 mmol, 3.0 equiv.) were dissolved in t-BuOH (4 mL). The mixture was degassed (N, 5 min) and then stirred at 90 °C for 3 h. The mixture was filtered through a plug of Celite, and the Celite was washed with EtOAc (30 mL). The filtrate was dry-loaded onto silica gel and purified by flash chromatography (silica gel, 12 g, 0–100% EtOAc, followed by isocratic 10% MeOH in DCM) to afford the title compound (32.0 mg, 31%) as a pale yellow solid. LCMS m / z 256.1[M+H]+,ESI pos.

[0157] Step C: 3-Hydroxy-4-[8-(3-hydroxy-3-methyl-cyclobutyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile The above (4-cyano-2-hydroxy-6-methyl-phenyl)boronic acid (33.2 mg, 0.19 mmol, 1.5 equiv.), 3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)-1-methyl-cyclobutanol (32.0 mg, 0.13 mmol, 1.0 equiv.), Xphos Pd G3 (15.91 mg, 0.02 mmol, 0.15 equiv.), and cesium carbonate (110.09 mg, 0.34 mmol, 2.7 equiv.) were suspended in 1,4-dioxane (3 mL) and water (0.75 mL) and degassed with N2 (5 min). The reaction mixture was heated to 80 °C and stirred for 3 h. The reaction mixture was filtered through a plug of Celite, and the filtrate was dry-loaded onto silica gel. The product was purified by flash chromatography (silica gel, 4 g column, 0-10% (0.7 N ammonia in MeOH / DCM)) to give the title compound (8.95 mg, 19% yield) as a light brown solid. LCMS m / z 352.8 [M+H]+, ESI pos.

[0158] Example 4: 3-Hydroxy-4-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile;2,2,2-Trifluoroacetic acid [ka] Step A: 3-hydroxy-5-methyl-4-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]benzonitrile To a solution of 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((2-(trimethylsilyl)ethoxy)methoxy)benzonitrile (106.5 mg, 0.27 mmol, 1.5 equiv; Intermediate 3), NaCO (57.9 mg, 0.55 mmol, 3.0 equiv), tert-butyl-[(1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethylsilane (70.0 mg, 0.18 mmol, 1.0 equiv) in 1,4-dioxane (1 mL) and water (0.2 mL) was added XphosPdG (15.5 mg, 0.02 mmol, 0.1 equiv). The reaction mixture was then stirred under N2 at 95°C for 2 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 5:1) to give the title compound (20.0 mg, 12% yield) as an orange oil. LCMS: m / z 611.5 [M+H] + ,ESI pos.

[0159] Step B: 3-hydroxy-4-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid A solution of 3-hydroxy-5-methyl-4-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]benzonitrile (20.0 mg, 0.03 mmol, 1.0 equiv.) and HCl / dioxane (0.5 mL, 1.0 mmol, 2 M in dioxane) was stirred at 25 °C for 10 min. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Gemini 150 mm × 30 mm × 25 μm; mobile phase: [water (0.1% TFA, V / V)-ACN]; B%: 22%–56%, 13 min) to give the title compound (8.6 mg, 70% yield) as a white solid. LCMS: m / z 367.1 [M+H] + ,ESI pos.

[0160] Example 5: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol;2,2,2-trifluoroacetic acid [ka] Step A: 5-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol 6-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indan-4-ol (5.4 mg, 0.02 mmol, 1.0 equiv.), tert-butyl-[(1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazolidinyl]-2-methyl-4-oxazolidinyl]-2-methyl-2- ... To a solution of [(methyl-8-yl)cyclohexoxy]-dimethyl-silane (10.0 mg, 0.02 mmol, 1.0 equiv.) and CsF (9.0 mg, 0.06 mmol, 3.0 equiv.) was added XPhosPdG3 (3.4 mg, 0.01 mmol, 0.2 equiv.) under N2, and the reaction vessel was then sealed and heated in a microwave at 95 °C for 1 h under N2. After the reaction was complete, the reaction mixture was cooled to 25 °C and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 2:1) to give the title compound (7.0 mg, 71% yield) as a yellow oil. LCMS: m / z 496.3 [M+H] + ,ESI pos.

[0161] Step B: 5-(8-((1R,2R)-2-hydroxycyclohexyl)-7,8-dihydro-6H-pyridazino[4,3-b][1,4]oxazin-3-yl)-6-methyl-2,3-dihydro-1H-indan-4-ol; 2,2,2-trifluoroacetic acid A solution of the aforementioned 5-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol (10.0 mg, 0.02 mmol, 1.0 equiv.) and HCl / dioxane (0.1 mL, 0.2 mmol, 2 M in dioxane) was stirred at 20° C. for 10 minutes. After the reaction was complete, the reaction mixture was concentrated in vacuo, and the residue was purified by preparative HPLC (column: Xtimate C). 18 , 250 mm × 50 mm × 10 μm; mobile phase: [water (0.1% TFA, v / v)-ACN]; B%: 5% to 55%, 10 min) to give the title compound (2.7 mg, 35% yield) as a white solid. LCMS: m / z 382.2 [M+H] +,ESI pos.

[0162] Example 6: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol;2,2,2-trifluoroacetic acid [ka] Step A: 5-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dimethylbenzofuran-4-ol To a solution of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (11.9 mg, 0.05 mmol, 1.2 equiv; CAS number 2923540-32-1), tert-butyl-[(1R,2R)-2-(3-chloro-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane (15.0 mg, 0.04 mmol, 1.0 equiv) and CsF (17.2 mg, 0.11 mmol, 3.0 equiv) in 1,4-dioxane (1 mL) and water (0.1 mL) was added XphosPdG (6.4 mg, 0.01 mmol, 0.2 equiv) under N. The reaction vessel was sealed and heated in a microwave at 95° C. for 0.5 hours under N2. After the reaction was completed, the reaction mixture was cooled to 20° C. and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate=5:1) to give the title compound (15.0 mg, 74% yield) as a yellow oil. LCMS: m / z 498.2 [M+H] + ,ESI pos.

[0163] Step B: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol; 2,2,2-trifluoroacetic acid A mixture of the aforementioned 5-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol (15.0 mg, 0.03 mmol, 1.0 equiv.) and HCl / dioxane (1.51 mL, 3.01 mmol, 2 M in dioxane) was stirred at 20°C for 1 minute. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Xtimate C). 18 , 250 mm × 50 mm × 10 μm; Mobile phase: [water (0.1% TFA, v / v)-ACN]; B%: 5% to 60%, 18 min to give the title compound (7.2 mg, 48% yield) as a white solid. LCMS: m / z 383.45 [M+H] + ,ESI pos.

[0164] Example 7: 4-[8-[(3R,5S)-1-ethyl-5-hydroxy-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile [ka] Step A: tert-butyl (3R,5S)-3-amino-5-[tert-butyl(dimethyl)silyl]oxy-piperidine-1-carboxylate To a mixture of tert-butyl (3R,5S)-3-amino-5-hydroxy-piperidine-1-carboxylate (CAS number 1932513-59-1, 550.0 mg, 2.54 mmol, 1 equiv.) and triethylamine (0.47 mL, 3.36 mmol, 1.32 equiv.) in DCM (10 mL) was added tert-butyldimethylsilyl chloride (421.6 mg, 2.8 mmol, 1.1 equiv.). The reaction mixture was stirred at 25° C. for 20 hours. After the reaction was complete, the reaction mixture was treated with ethyl acetate (15 mL) and a mixture (30 mL) of water / brine (1:1, V / V). The aqueous layer was extracted with ethyl acetate (50 mL × 3). The organic layer was washed with brine (60 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:5 to 0:1) to give the title compound (380.0 mg, yield 45%) as a yellow oil. 1 H NMR(400MHz,CD3OD)δ[ppm]:3.89-3.77(m,2H),3.77-3.70(m,1H),2.96-2.74(m, 3H),2.16-2.06(m,1H),1.46(s,9H),1.44-1.38(m,1H),0.92(s,9H),0.12(d,6H).

[0165] Step B: tert-butyl (3S,5R)-3-[tert-butyl(dimethyl)silyl]oxy-5-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]piperidine-1-carboxylate To a solution of 2-(3,6-dichloropyridazin-4-yl)oxyacetaldehyde (540.0 mg, 1.3 mmol, 1.0 equiv) in DCE (11 mL) was added tert-butyl (3R,5S)-3-amino-5-[tert-butyl(dimethyl)silyl]oxy-piperidine-1-carboxylate (340.0 mg, 1.03 mmol, 0.79 equiv). The mixture was then stirred at 20 °C for 0.5 h. Then, NaBH(OAc) (691.1 mg, 3.26 mmol, 2.5 equiv) was added portionwise to the above mixture, and the resulting mixture was stirred at 20 °C for 0.5 h. After the reaction was complete, the reaction mixture was cooled to room temperature and quenched with water (30 mL). The aqueous phase was extracted with DCM (30 mL × 2). The combined phase was washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1 to 1:3) to give the title compound (200.0 mg, 29% yield) as a yellow oil. LCMS: m / z 521.2 [M+H] + ,ESI pos.

[0166] Step C: tert-butyl (3S,5R)-3-[tert-butyl(dimethyl)silyl]oxy-5-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)piperidine-1-carboxylate To a solution of tert-butyl (3S,5R)-3-[tert-butyl(dimethyl)silyl]oxy-5-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]piperidine-1-carboxylate (350.0 mg, 0.67 mmol, 1.0 equiv.) and CsCO (437.3 mg, 1.34 mmol, 2.0 equiv.) in 1,4-dioxane (6 mL), Pd-PEPPSI-IHEPTCl (CAS No. 1814936-54-3, 56.4 mg, 0.07 mmol, 0.1 equiv.) was added. The mixture was then stirred at 80° C. for 12 hours under N. After the reaction was complete, the reaction mixture was cooled to 20° C. The mixture was then concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:1 to 0:1) to give the title compound (140.0 mg, 22% yield) as a yellow oil. LCMS: m / z 485.1 [M+H] + ,ESI pos.

[0167] Step D: tert-butyl (3S,5R)-3-[tert-butyl(dimethyl)silyl]oxy-5-[3-[4-cyano-2-methyl-6-(2-trimethylsilylethoxymethoxy)phenyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]piperidine-1-carboxylate To a solution of 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(2-trimethylsilylethoxymethoxy)benzonitrile (60.1 mg, 0.15 mmol, 1.2 equiv.), tert-butyl (3S,5R)-3-[tert-butyl(dimethyl)silyl]oxy-5-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8yl)piperidine-1-carboxylate (120.0 mg, 0.13 mmol, 1.0 equiv.) and CsF (78.2 mg, 0.51 mmol, 4.0 equiv.) in 1,4-dioxane (2 mL) and water (0.4 mL) was added XphosPdG (12.0 mg, 0.01 mmol, 0.11 equiv.) under N. The reaction vessel was sealed and heated in a microwave at 95°C for 30 minutes. After the reaction was completed, the reaction mixture was cooled to 20°C and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1) to give the title compound (60.0 mg, 47% yield) as a yellow oil. LCMS: m / z 712.4 [M+H] + ,ESI pos.

[0168] Step E: 3-hydroxy-4-[8-[(3R,5S)-5-hydroxy-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid A solution of tert-butyl (3S,5R)-3-[tert-butyl(dimethyl)silyl]oxy-5-[3-[4-cyano-2-methyl-6-(2-trimethylsilylethoxymethoxy)phenyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]piperidine-1-carboxylate (50.0 mg, 0.05 mmol, 1.0 equiv.) in TFA (2.03 mL, 27.31 mmol) was stirred at 40° C. for 16 hours. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by reverse-phase flash chromatography (Xtimate C column). 18, 250 mm × 50 mm × 10 μm; Mobile phase: [water (0.1% TFA, v / v)-ACN]; B%: 5% to 40%, 13 min) to give the title compound (20.0 mg, 82% yield) as a yellow solid. LCMS: m / z 368.2 [M+H] + ,ESI pos.

[0169] Step F: 4-[8-[(3R,5S)-1-ethyl-5-hydroxy-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile 3-Hydroxy-4-[8-[(3R,5S)-5-hydroxy-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; 2,2,2 trifluoroacetic acid (15.0 mg, 0.03 mmol, 1.0 equiv.) and DIEA (10.1 mg, 0.08 mmol, 2.5 equiv.) were added to iodoethane (5.8 mg, 0.04 mmol, 1.2 equiv.) in DMF (1 mL). The reaction mixture was then stirred at 20 °C for 1 h. After the reaction was completed, water (0.1 mL) was added, and the mixture was then purified by preparative HPLC (column: Phenomenex Gemini, 150 mm x 25 mm x 10 μm; mobile phase: [water (0.1% NH4HCO3, V / V)-ACN]; B%: 22%-52%, 12 min) to give the title compound (5.47 mg, 45% yield) as a white solid. LCMS: m / z 396.2 [M+H] + ,ESI pos.

[0170] Example 8: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol [ka] Step A: 3,6-Dichloro-4-(2-iodoethoxy)pyridazine 2-Iodoethanol (107.0 μL, 1.37 mmol, 1.01 equiv) was dissolved in anhydrous THF (7 mL) and sodium hydride (60% in mineral oil) (82.0 mg, 2.05 mmol, 1.5 equiv) was added portionwise at 0° C. The mixture was stirred at room temperature for 15 minutes, then 3,4,6-trichloropyridazine (250.0 mg, 1.36 mmol, 1.0 equiv) was added and the mixture was stirred at 40° C. overnight (18 h). The reaction was left stirring for an additional day. The reaction mixture was quenched with water (100 mL) and extracted with EtOAc (2×100 mL). The combined organic extracts were dried (NaSO) and concentrated in vacuo. The resulting residue was purified by flash chromatography on silica gel (40 g column, 0-40% EtOAc in heptane) eluting with 30% to afford the title compound (88.0 mg, 20% yield) as a pale yellow solid. LCMS: m / z 318.9 2x 35 Cl[M+H] + ,ESI pos.

[0171] Step B: (1R,2R)-2-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]cyclohexanol 3,6-Dichloro-4-(2-iodoethoxy)pyridazine (418.0 mg, 1.31 mmol, 1.0 equiv.), (1R,2R)-2-aminocyclohexanol (227.0 mg, 1.97 mmol, 1.5 equiv.), and N,N-diisopropylethylamine (0.9 mL, 5.17 mmol, 3.94 equiv.) were dissolved in DMSO (8 mL) and stirred at 50 °C for 2 h. The reaction was allowed to react overnight (16 h). After this time, the reaction mixture was diluted with EtOAc (50 mL) and 50 wt.% brine (50 mL), and the layers were separated. The aqueous phase was extracted with EtOAc (2 × 50 mL), and the combined organics were dried (NaSO), filtered, and concentrated in vacuo. The crude product was combined with another batch and purified by silica gel chromatography (40 g cartridge, 0-10 (MeOH: EtOAc) eluting with 10% to give the title compound (124.0 mg, 28% yield) as a pale yellow solid. LCMS: m / z 306.1 [M+H] +,ESI pos.

[0172] Step C: (1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexanol Pd-176 (360.0 mg, 0.45 mmol, 0.17 equiv.) and cesium carbonate (2.6 g, 7.98 mmol, 2.96 equiv.) were added to a flask containing (1R,2R)-2-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]-cyclohexanol (825.0 mg, 2.69 mmol, 1.0 equiv.) in sparged tert-butanol (35 mL) (sonicated for 5 min while bubbling N). The reaction mixture was heated to 80 °C and left stirring for 2 h. The mixture was combined with other batches and filtered through a plug of Celite, which was washed with EtOAc (30 mL). The concentrated filtrate (1.69 g) was dry loaded onto Celite and purified by reverse-phase flash chromatography (C) eluting with 25% HCl. 18 Purification by HPLC (43 g column, 10-100% MeCN [0.1% formic acid]:0.1% formic acid in water) and extraction of the appropriate fractions with DCM (3 x 50 mL) followed by drying (Na2SO4), filtration and concentration afforded the title compound (76.0 mg, 10% yield) as a pale yellow solid. LCMS: m / z 270.1 / 272.1 [M+H] + ,ESI pos.

[0173] Step D: (1R,2R)-2-[3-(4-benzyloxy-6-methyl-2,3-dihydrobenzofuran-5-yl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]cyclohexanol A mixture of 2-(4-benzyloxy-6-methyl-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS number 2923540-0, 82.0 mg, 0.22 mmol, 1.59 equiv.), (1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexanol (38.0 mg, 0.14 mmol, 1.0 equiv.), and cesium carbonate (136.0 mg, 0.42 mmol, 2.96 equiv.) in water (0.400 mL) and 1,4-dioxane (2 mL) was degassed with N (sonication, 5 min). XPhos Pd G3 (18.0 mg, 0.02 mmol, 0.15 equiv) was added, and the reaction mixture was heated to 80 °C and stirred for 4 h. The reaction was cooled and stirred over the weekend, but no change was observed. Additional 2-(4-benzyloxy-6-methyl-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (27.0 mg, 0.07 mmol, 0.52 equiv) and cesium carbonate (49.0 mg, 0.15 mmol, 1.07 equiv) were added, and the reaction mixture was sparged as before. XPhos Pd G3 (12.0 mg, 0.01 mmol, 0.1 equiv) was then added, and the reaction mixture was heated as before for 2 h. The reaction mixture was dry-loaded onto silica gel. The product was purified by flash chromatography on silica gel (24 g column, 0-10% MeOH:EtOAc) eluting with 5% to give the title compound (13.0 mg, 18% yield) as a pale yellow solid. LCMS: m / z 474.3 [M+H] + ,ESI pos.

[0174] Step E: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol Pd / C (type 87) (16.0 mg, 0.01 mmol, 0.1 equiv.) was added to a stirred solution of (1R,2R)-2-[3-(4-benzyloxy-6-methyl-2,3-dihydrobenzofuran-5-yl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]cyclohexanol (36.0 mg, 0.08 mmol, 1.0 equiv.) in ethanol (3 mL). The hydrogenation vessel was placed under an atmosphere of hydrogen (2 bar) at room temperature and stirred vigorously for 2 h. The reaction mixture was filtered through Celite, rinsed with EtOH, and concentrated under reduced pressure to give a crude residue (34 mg). The crude material was dissolved in 1.58 mL of DMSO, filtered, and purified by Waters X-Select CSH C 18 Purification was performed by reverse-phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organizer, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) using an ODB preparative column, 130 Å, 5 μm, 30 mm × 100 mm, at a flow rate of 40 mL / min, eluting with a 0.1% formic acid in water-MeCN gradient over 8.5 min using UV across all wavelengths with PDA, QDa, and ELS detectors. An at-column dilution pump provided 2 mL / min of methanol throughout the method, which is included in the MeCN percentages below. Gradient information: 0.0-0.5 min, 12.5% ​​MeCN; 0.5-5.5 min, ramp from 12.5% ​​MeCN to 42.5% MeCN; 5.5-5.6 min, ramp from 42.5% MeCN to 100% MeCN; 5.6-8.5 min, hold at 100% MeCN. Clean fractions were evaporated in a Genevac. The dried fractions were transferred to a vial and lyophilized to afford the title compound (16.0 mg, 53% yield) as an off-white amorphous lyophilized solid. LCMS: m / z 384.2 [M+H] + ,ESI pos.

[0175] Example 9: 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethyl)phenol [ka] 3-Methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(trifluoromethyl)phenol (CAS number 2557358-8, 66.0 mg, 0.22 mmol, 1.55 equiv.), (1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexanol (38.0 mg, 0.14 mmol, 1.0 equiv.; Example 8, Step C), and cesium carbonate (131.0 mg, 0.4 mmol, 2.85 equiv.) in water (0.400 mL) and 1,4-dioxane (2 mL) were degassed with N. XPhos Pd G3 (18.0 mg, 0.02 mmol, 0.15 equiv) was added, and the reaction mixture was heated to 80 °C and stirred for 3 h. The reaction mixture was dry-loaded onto silica gel. The product was purified by flash chromatography on silica gel (0-60% (10% MeOH [0.7 M NH3] in DCM):DCM) eluting at 45% to give the product as a yellow gum with some minor impurities. The residue was triturated with TBME and PE 40:60. The resulting precipitate was filtered and dried for 24 h to give the title compound (24.0 mg, 39% yield) as a yellow solid. LCMS: m / z 410.2 [M+H] + ,ESI pos.

[0176] Example 10: 4-[8-[(3R)-1-Ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid [ka] Step A: tert-butyl (3R)-3-[2-(3,6-dihydropyridazin-4-yl)oxyethylamino]pyrrolidine-1-carboxylate To a solution of 2-(3,6-dichloropyridazin-4-yl)oxyethyl ethanesulfonate (2600.0 mg, 8.63 mmol, 1.0 equiv.) and (R)-(+)-1-BOC-3-aminopyrrolidine (3216.1 mg, 17.27 mmol, 2.0 equiv.) in DMSO (13 mL), DIEA (1670.6 mg, 12.95 mmol, 1.5 equiv.) was added. The reaction mixture was then stirred at 80° C. for 12 hours. After the reaction was complete, the mixture was poured into water (30 mL) and extracted with EtOAc (200 mL×2). The combined organic phase was washed with brine (100 mL×2), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=0:1 to 1:1) to give the title compound (2.0 g, yield 61%) as a yellow solid. 1 H NMR(400MHz,CDCl3)δ[ppm]:7.06(s,1H),4.45-4.32(m,1H),3.78-3.65(m,3H),3.63 -3.51(m,3H),3.39-3.23(m,2H),2.26-2.13(m,1H),2.05-1.95(m,1H),1.48(s,9H).

[0177] Step B: tert-Butyl (3R)-3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)pyrrolidine-1-carboxylate To a solution of tert-butyl (3R)-3-[2-(3,6-dichloropyridazin-4-yl)oxyethylamino]pyrrolidine-1-carboxylate (2.0 g, 5.3 mmol, 1.0 equiv.), BINAP (429.1 mg, 0.69 mmol, 0.13 equiv.), and CsCO (3281.7 mg, 10.07 mmol, 1.9 equiv.) in toluene (20 mL) was added Pd(OAc) (95.2 mg, 0.42 mmol, 0.08 equiv.) at 25 °C under N. The reaction mixture was then stirred at 110 °C for 2 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine (20 mL × 2), dried over sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 5:1) to give the title compound (160.0 mg, 9% yield) as a yellow solid. LCMS: m / z 341.1 [M+H] + ,ESI pos.

[0178] Step C: 3-chloro-8-[(3R)-pyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazine A solution of tert-butyl (3R)-3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)pyrrolidine-1-carboxylate (160.0 mg, 0.47 mmol, 1.0 equiv.) in DCM (1 mL) and TFA (0.5 mL) was stirred at 25° C. for 0.5 h. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by reverse-phase flash chromatography (column: Xtimate C). 18 , 250 mm × 50 mm × 10 μm; mobile phase: [water (0.1% ammonium hydroxide v / v)-ACN]; B%: 5% to 35%, 10 min) to give the title compound (30.0 mg, 27% yield) as a yellow oil. LCMS: m / z 241.0 [M+H] + ,ESI pos.

[0179] Step D: 3-chloro-8-[(3R)-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazine To a solution of 3-chloro-8-[(3R)-pyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazine (30.0 mg, 0.12 mmol, 1.0 equiv.), DIEA (32.16 mg, 0.25 mmol, 2.0 equiv.) in DMF (1 mL) was added iodoethane (0.01 mL, 0.19 mmol, 1.5 equiv.). The mixture was then stirred at 20°C for 1 hour. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by reverse-phase flash chromatography (Xtimate C column). 18 , 250 mm × 40 mm × 10 μm; mobile phase: [water (0.1% ammonium hydroxide v / v)-MeCN]; B%: 5% to 38%, 15 min), followed by lyophilization to give the title compound (22.0 mg, 66% yield) as a yellow solid. LCMS: m / z 269.1 [M+H] + ,ESI pos.

[0180] Step E: 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(2-trimethylsilylethoxymethoxy)benzonitrile To a mixture of CsF (37.3 mg, 0.25 mmol, 3.0 equiv.), 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((2-(trimethylsilyl)ethoxy)methoxy)benzonitrile (47.8 mg, 0.12 mmol, 1.5 equiv., Intermediate 3), and 3-chloro-8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazine (22.0 mg, 0.08 mmol, 1.0 equiv.) in 1,4-dioxane (1 mL) and water (0.2 mL) was added XPhos Pd G3 (6.9 mg, 0.01 mmol, 0.1 equiv.) under N2 at 25 °C. The mixture was stirred at 95 °C for 2.5 h. After the reaction was complete, the reaction mixture was cooled to room temperature. EtOAc (30 mL) and brine (20 mL) were added to the mixture, and the layers were separated. The aqueous phase was extracted with EtOAc (30 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1) to give the title compound (21.0 mg, 42% yield) as a yellow solid. 1 H NMR(400MHz,CD3OD)δ[ppm]:7.54(s,1H),7.42(s,1H),7.09(s,1H),5.36(s,2H),5.01-5.00(m,1H),4.78-4.75(m,2H),4.05-4.00(m,1H),3 .82-3.80(m,2H),3.67(t,2H),3.35-3.34(m,3H),3.27-3.18(m,1H),2 .85-2.70(m,2H),2.22(s,3H),1.39(t,3H),0.91(t,2H),0.00(s,9H).

[0181] Step F: 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(2-trimethylsilylethoxymethoxy)benzonitrile (21.0 mg, 0.03 mmol, 1.0 equiv.) in DCM (1 mL) and TFA (0.5 mL) was stirred at 25 °C for 1 h. After the reaction was complete, the reaction mixture was concentrated in vacuo, dissolved in MeOH (2 mL), and basified to pH = 7 with NH3·H2O. The mixture was then filtered, and the filtrate was concentrated in vacuo. The residue was purified by preparative HPLC (column: Xtimate C 18 , 250 mm × 50 mm × 10 μm; mobile phase: [water (0.1% TFA, v / v)-ACN]; B%: 5% to 35%, 10 min) followed by lyophilization to give the title compound (10.98 mg, 62% yield) as a pale yellow solid. LCMS: m / z 366.1 [M+H] + ,ESI pos.

[0182] Example 11: 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol; 2,2,2-trifluoroacetic acid [ka] Step A: 1-benzyloxy-2-bromo-3,5-dimethyl-benzene To a solution of 2-bromo-3,5-dimethyl-phenol (5.0 g, 24.9 mmol, 1.0 equiv.), bromomethylbenzene (5.1 g, 29.8 mmol, 1.2 equiv.) in DMF (50 mL) was added CsCO (16.2 g, 49.7 mmol, 2.0 equiv.). The reaction mixture was stirred at 25 °C under a nitrogen atmosphere for 1 h. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL × 3). The combined organic phase was washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica flash chromatography (petroleum ether) to give the title compound (7.0 g, 87% yield) as a colorless oil. 1 H NMR(400MHz,DMSO-d6)δ[ppm]:7.54-7.28(m,5H),6.90-6.83(m,1H),6.80-6.74(m,1H),5.16(s,2H),2.30(s,3H),2.24(s,3H).

[0183] Step B: 2-(2-benzyloxy-4,6-dimethyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane To a solution of 1-benzyloxy-2-bromo-3,5-dimethylbenzene (7.0 g, 24.04 mmol, 1.0 equiv.) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (12.21 g, 48.08 mmol, 2.0 equiv.) in 1,4-dioxane (70 mL), CsCO (15.67 g, 48.08 mmol, 2.0 equiv.), tris(4-methoxy-3,5-dimethylphenyl)phosphane (1.05 g, 2.4 mmol, 0.1 equiv.) and Pd(OAc) (0.54 g, 2.4 mmol, 0.1 equiv.) were added. The mixture was then stirred at 95 °C under N for 2 h. The reaction mixture was cooled to room temperature, then diluted with water (300 mL) and ethyl acetate (200 mL), filtered, and the filtrate was extracted with ethyl acetate (200 mL × 3). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 0:1) to give the title compound (3.5 g, 39% yield) as a colorless oil. 1 H NMR(400MHz,DMSO-d6)δ[ppm]:7.66-7.20(m,5H),6.68(s,1H),6.55(s,1H),5.01(s,2H),2.25(s,3H),2.22(s,3H),1.22(s,12H).

[0184] Step C: 3,5-dimethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol Pd / C (0.26 g, 0.22 mmol, 0.11 equiv. purity: 10%) in EtOAc (4 mL) was added to a solution of 2-(2-benzyloxy-4,6-dimethyl-phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.65 g, 1.92 mmol, 1.0 equiv.) in EtOAc (1 mL). The reaction mixture was then stirred under H (1100 mmHg) at 25° C. for 2 hours. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure to give the title compound (420.0 mg, 79% yield) as an orange oil.1 H NMR(400MHz,CD3OD)δ[ppm]:6.49(s,1H),6.43(s,1H),2.38(s,3H),2.21(s,3H),1.38(s,12H).

[0185] Step D: 2-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol To a mixture of tert-butyl-[(1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane (50.0 mg, 0.13 mmol, 1.0 equiv.), NaCO (41.4 mg, 0.39 mmol, 3.0 equiv.), and 3,5-dimethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (48.5 mg, 0.2 mmol, 1.5 equiv.) in 1,4-dioxane (2.5 mL) and water (0.5 mL) was added XphosPdG (22.1 mg, 0.03 mmol, 0.2 equiv.). The reaction mixture was stirred at 95 °C under N for 1.5 h. The reaction mixture was cooled to room temperature. The reaction mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by preparative TLC (petroleum ether:ethyl acetate=3:1) to give the title compound (40.0 mg, 31% yield) as an orange oil. LCMS: m / z 470.4 [M+H] + ,ESI pos.

[0186] Step E: 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol; 2,2,2-trifluoroacetic acid A solution of 2-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol (20.0 mg, 0.04 mmol, 1.0 equiv.) and HCl / dioxane (0.65 mL, 1.3 mmol, 2 M in dioxane) was stirred at 25 °C for 10 minutes. The mixture was concentrated under vacuum. The residue was purified by preparative HPLC (column: Phenomenex Gemini 150 mm × 30 mm × 15 μm; mobile phase: [water (0.1% TFA, V / V)-ACN]; B%: 1% to 55%, 12 min) to give the title compound (19.4 mg, 49% yield) as a white solid. LCMS: m / z 356.1 [M+H] + ,ESI pos.

[0187] Example 12: 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol;2,2,2-Trifluoroacetic acid [ka] Step A: 3-methyl-5-(trifluoromethoxy)aniline To a solution mixture of 3-bromo-5-(trifluoromethoxy)aniline (10.0 g, 39.1 mmol, 1.0 equiv.) and trimethylboroxine (16.7 mL, 58.6 mmol, 1.5 equiv.) in 1,4-dioxane (100 mL) and water (20 mL), K2CO3 (10.8 g, 78.1 mmol, 2.0 equiv.) was added, and the mixture was degassed and purged with N2 three times. Pd(dppf)Cl2 (1.43 g, 1.95 mmol, 0.05 equiv.) was then added to the mixture. The mixture was stirred at 100 °C under N2 for 12 h. After the reaction was complete, the reaction mixture was cooled to room temperature. EtOAc (150 mL) and water (50 mL) were added to the mixture, and the layers were separated. The aqueous phase was extracted with EtOAc (200 mL × 2). The combined extracts were washed with brine (100 mL x 2), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1 to 5:1) to give the title compound (6.2 g, 56% yield) as a yellow oil. LCMS: m / z 192.2 [M+H] + ,ESI pos

[0188] Step B: 3-methyl-5-(trifluoromethoxy)phenol To a mixture of 3-methyl-5-(trifluoromethoxy)aniline (7.2 g, 37.7 mmol, 1.0 equiv.) in sulfuric acid (35.0 mL) / water (35 mL), NaNO (5.2 g, 75.33 mmol, 2.0 equiv.) was added, purged with N three times, and the mixture was stirred at 80 °C for 3 h. After the reaction was complete, the mixture was poured into water (50 mL) at 0 °C. EtOAc (100 mL) was added to the above mixture, and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL × 2). The combined phase was washed with brine (50 mL × 2), dried over Na SO , filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether / ethyl acetate = 10 / 0 to 5 / 1) to give the title compound (1.3 g, 16% yield) as a brown oil. LCMS: m / z 191.2 [MH] - ,ESI neg.

[0189] Step C: 2-iodo-3-methyl-5-(trifluoromethoxy)phenol To a mixture of 3-methyl-5-(trifluoromethoxy)phenol (3.2 g, 16.65 mmol, 1.0 equiv.) in toluene (60 mL) under N2, NaH (1.33 g, 33.3 mmol, 2.0 equiv. 60% in mineral oil) was added, and the reaction mixture was then stirred for 30 min. I2 (4.23 g, 16.65 mmol, 1.0 equiv.) was added to the above mixture. The mixture was stirred at 25 °C for 16 h. After the reaction was complete, the reaction mixture was quenched with water (10 mL) and extracted with EtOAc (20 mL). The aqueous phase was extracted with EtOAc (30 mL × 2). The combined phase was washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 100:1) to give the title compound (2.3 g, 39% yield) as a yellow oil. LCMS: m / z 317.0 [M+H] + ,ESI pos.

[0190] Step D: 1-(benzyloxy)-2-iodo-3-methyl-5-(trifluoromethoxy)benzene To a mixture of 2-iodo-3-methyl-5-(trifluoromethoxy)phenol (1.27 g, 3.59 mmol, 1.0 equiv.) in DMF (10 mL) was added K2CO3 (0.99 g, 7.19 mmol, 2.0 equiv.) and BnBr (0.85 g, 7.16 mmol, 1.1 equiv.). The reaction mixture was stirred at 25 °C for 16 h. After the reaction was complete, the mixture was quenched with 1 N HCl (10 mL). EtOAc (40 mL) and water (40 mL) were added to the above mixture, and the layers were separated. The aqueous phase was extracted with EtOAc (30 mL × 2). The combined phase was washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=1:0 to 10:1) to give the title compound (1.2 g, yield 78%) as a colorless oil. 1H NMR(400MHz,CD3OD)δ[ppm]:7.52(d,2H),7.41-7.35(m,2H),7.33-7.29(m,1H),6.88(s,1H),6.75(d,1H),5.17(s,2H),2.49(s,3H).

[0191] Step E: 2-[2-benzyloxy-6-methyl-4-(trifluoromethoxy)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane To a solution of 1-benzyloxy-2-iodo-3-methyl-5-(trifluoromethoxy)benzene (1.10 g, 2.7 mmol, 1.0 equiv.) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.37 g, 5.39 mmol, 2.0 equiv.) in 1,4-dioxane (20 mL), CsCO (1.76 g, 5.39 mmol, 2.0 equiv.), tris(4-methoxy-3,5-dimethylphenyl)phosphane (235.3 mg, 0.54 mmol, 0.2 equiv.) was added Pd(OAc) (121.0 mg, 0.54 mmol, 0.2 equiv.) under N at 25 °C. The mixture was then stirred at 95 °C under N for 4 h. After the reaction was completed, the reaction mixture was cooled to room temperature. EtOAc (50 mL) and water (20 mL) were added to the mixture, and the layers were separated. The aqueous phase was extracted with EtOAc (50 mL × 2). The combined phases were washed with brine (20 mL × 2), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 10:1) to give the title compound (620.0 mg, 54% yield) as a white solid. 1 H NMR(400MHz,CD3OD)δ[ppm]:7.48(d,2H),7.39-7.34(m,2H),7.33-7.29(m,1H),6.71(s,1H),6.68(s,1H),5.03(s,2H),2.35(s,3H),1.28(s,12H).

[0192] Step F: [(1R,2R)-2-[3-[2-benzyloxy-6-methyl-4-(trifluoromethoxy)phenyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]cyclohexoxy]-tert-butyl-dimethyl-silane To a solution of 2-[2-benzyloxy-6-methyl-4-(trifluoromethoxy)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (31.9 mg, 0.08 mmol, 1.5 equiv.), NaCO (16.6 mg, 0.16 mmol, 3.0 equiv.), and tert-butyl-[(1R,2R)-2-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane (20.0 mg, 0.05 mmol, 1.0 equiv.) in 1,4-dioxane (1 mL) / water (0.20 mL) was added XphosPdG (8.8 mg, 0.01 mmol, 0.2 equiv.) under N at 25 °C. The mixture was stirred at 95 °C for 16 h. After the reaction was completed, the reaction was cooled to room temperature. EtOAc (10 mL) and water (5 mL) were added to the above mixture, and the layers were separated. The aqueous phase was extracted with EtOAc (10 mL x 2). The combined phase was washed with brine (10 mL x 2), dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 5:1) to give the title compound (12.0 mg, 35% yield) as a yellow solid. LCMS: m / z 630.4 [M+H] + ,ESI pos.

[0193] Step G: 2-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol To a solution of Pd / C (10.0 mg) in EtOAc (1 mL) and methanol (0.1 mL) under N was added [(1R,2R)-2-[3-[2-benzyloxy-6-methyl-4-(trifluoromethoxy)phenyl]-6,7-dihydropyridazino[4,3b][1,4]oxazin-8-yl]cyclohexoxy]-tert-butyl-dimethyl-silane (70.0 mg, 0.11 mmol, 1.0 equiv.). The reaction mixture was stirred under H (1100 mmHg) at 25 °C for 1 h. After the reaction was complete, the suspension was filtered through a pad of Celite, and the filter cake was washed with EtOAc (20 mL × 4). The combined filtrate was concentrated under vacuum to give the title compound (60.0 mg, 85% yield) as a yellow oil. LCMS: m / z 540.3 [M+H] + ,ESI pos.

[0194] Step H: 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol; 2,2,2-trifluoroacetic acid A solution of 2-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol (50.0 mg, 0.09 mmol, 1.0 equiv.) and HCl / dioxane (0.5 mL, 1.0 mmol, 2 M in dioxane) was stirred at 25 °C for 10 min. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by preparative HPLC (Phenomenex Gemini 150 mm × 30 mm × 15 μm column; mobile phase: [water (0.1% TFA, V / V)-ACN]; B%: 3%–53%, 12 min) to give the title compound (33.1 mg, 65% yield) as a white solid. LCMS: m / z 426.2 [M+H] + ,ESI pos.

[0195] Example 13: 4-[8-[(3R)-1-Ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxybenzonitrile; 2,2,2-trifluoroacetic acid [ka] Step A: 3-chloro-8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazine To a solution of 2-(3,6-dichloro-5-methyl-pyridazin-4-yl)oxyethyl ethanesulfonate (220.0 mg, 0.7 mmol, 1.0 equiv.), (3R)-1-ethylpiperidin-3-amine (116.3 mg, 0.91 mmol, 1.3 equiv.) in DMSO (1 mL) was added DIEA (162.1 mg, 1.26 mmol, 1.8 equiv.) under N2. The reaction mixture was then stirred at 80°C for 16 hours. After the reaction was complete, the reaction mixture was cooled to 25°C. The solution was purified by reverse-phase flash (column: Xtimate C18, 250 mm x 50 mm x 10 μm; mobile phase: [water (0.1% FA, v / v)-ACN]; B%: 5%-35%, 10 min), followed by lyophilization to give the title compound (18.0 mg, 8.32% yield) as a yellow solid. LCMS: m / z 297.2 [M+H] + ,ESI pos.

[0196] Step B: 4-[8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-(2-trimethylsilylethoxymethoxy)benzonitrile To a solution of 3-chloro-8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazine (18.0 mg, 0.06 mmol, 1.0 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(2-trimethylsilylethoxymethoxy)benzonitrile (34.1 mg, 0.09 mmol, 1.5 equiv.), and CsF (27.5 mg, 0.18 mmol, 3.0 equiv.) in dioxane (1.0 mL) and water (0.2 mL) was added XphosPdG3 (5.1 mg, 0.01 mmol, 0.1 equiv.). The reaction mixture was then stirred at 95° C. under nitrogen for 4 hours. The reaction mixture was cooled to room temperature. The mixture was concentrated in vacuo. The residue was purified by reversed-phase flash chromatography (column: Xtimate C 18 , 250 mm × 50 mm × 10 μm; mobile phase: [water (0.1% ammonium hydroxide v / v)-ACN]; B%: 5% to 35%, 10 min) followed by lyophilization to give the title compound (11.0 mg, 34% yield) as a yellow solid. LCMS: m / z 510.5 [M+H] + ,ESI pos.

[0197] Step C: 4-[8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-benzonitrile; 2,2,2-trifluoroacetic acid A solution of 4-[8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-(2-trimethylsilylethoxymethoxy)benzonitrile (11.0 mg, 0.02 mmol, 1.0 equiv.), TFA (104.7 mg, 1.08 mmol, 50.0 equiv.) in DCM (1 mL) was stirred under nitrogen at 25° C. for 1 hour. After the reaction was complete, the reaction mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Xtimate C). 18, 250 mm × 50 mm × 10 μm; mobile phase: [water (0.1% TFA, v / v)-ACN]; B%: 10% to 100%, 15 min) followed by lyophilization to give the title compound (5.3 mg, 50% yield) as a yellow solid. LCMS: m / z 380.2 [M+H] + ,ESI pos.

[0198] Example 14: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol;2,2,2-trifluoroacetic acid [ka] Step A: 5-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol To a solution of Intermediate 6 tert-butyl-[(1R,2R)-2-(3-chloro-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane (10.0 mg, 0.03 mmol, 1.0 equiv) in 1,4-dioxane (1 mL) / water (0.200 mL) was added NaCO (7.99 mg, 0.08 mmol, 3.0 equiv), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indan-4-ol (9.8 mg, 0.04 mmol, 1.5 equiv), and Xphos Pd g (4.26 mg, 0.01 mmol, 0.2 equiv) and the mixture was heated at 95 °C for 1.5 h. The reaction mixture was then cooled to room temperature, combined with another batch, and concentrated under reduced pressure. The crude product was purified by preparative TLC (petroleum ether:ethyl acetate=5:1, R f =0.5) to give the title compound (10.0 mg, 71% yield) as an orange oil. LCMS m / z: 496.2 [M+H] + ,ESI pos.

[0199] Step B: 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol; 2,2,2-trifluoroacetic acid To a solution of 5-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol (5.0 mg, 0.01 mmol, 1.0 equiv) in 1,4-dioxane (0.5 mL) was added dioxane / HCl (0.5 mL, 1.0 mmol, 99 equiv) and the mixture was stirred at 25 °C for 0.08 h. The mixture was then concentrated in vacuo and the crude product was purified by preparative HPLC (Column Phenomenex Luna C18 150 x 25 mm x 10 um Condition water (TFA)-ACN Begin B 20 End B 50 Gradient Time (min) 9 100% B Hold Time (min) 2 Flow Rate (ml / min) 25) to give the title compound as a white solid. The product was combined with another batch. (4.2 mg, 83% yield). LCMS m / z: 382.2 [M+H] + ,ESI pos.

[0200] Example 15: 3-[8-[(1R,2R)-2-Hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol; 2,2,2-Trifluoroacetic acid [ka] Step A: 3,6-Dichloro-4-(2-iodoethoxy)-5-methyl-pyridazine To a solution of 2-(3,6-dichloro-5-methyl-pyridazin-4-yl)oxyethyl ethanesulfonate (1.45 g, 4.6 mmol, 1.0 equiv.) in MeCN (15 mL) was added NaI (1.03 g, 6.9 mmol, 1.5 equiv.), and the reaction mixture was stirred at 80 °C for 1 h. The reaction mixture was cooled to room temperature, diluted with water (100 mL), and extracted with ethyl acetate (100 mL × 3). The combined organic phase was washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 2:1) to afford the title compound (1.0 g, 59% yield) as a yellow oil. 1 H NMR(400MHz,CD3OD)δ[ppm]:4.46(t,2H),3.57(t,2H),2.47(s,3H).

[0201] Step B: tert-Butyl-[(1R,2R)-2-(3-chloro-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane To a solution of 3,6-dichloro-4-(2-iodoethoxy)-5-methyl-pyridazine (500.0 mg, 1.5 mmol, 1.0 equiv.) in DMF (10 mL) was added DIEA (0.49 mL, 3.0 mmol, 2.0 equiv.) and trans-(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexanamine (516.8 mg, 2.25 mmol, 1.5 equiv.), and the mixture was stirred at 50° C. for 12 hours. The reaction mixture was cooled to room temperature, diluted with water (50 mL), and extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by preparative TLC (PE:EA 5:1) to afford the title compound (15.0 mg, 2% yield) as a yellow oil. LCMS: m / z 398.2 [M+H] + ,ESI pos.

[0202] Step C: [(1R,2R)-2-[3-(2-benzyloxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]cyclohexoxy]-tert-butyl-dimethyl-silane To a solution of tert-butyl-[(1R,2R)-2-(3-chloro-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane (15.0 mg, 0.02 mmol, 1.0 equiv.) in 1,4-dioxane (2 mL) / water (0.4 mL) was added Na2CO3 (5.9 mg, 0.06 mmol, 3. 0 equiv.), 2-(2-benzyloxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (19.0 mg, 0.03 mmol, 1.5 equiv.), and XphosPdG3 (6.4 mg, 0.01 mmol, 0.2 equiv.) were added, followed by stirring at 95°C for 1.5 h under a N2 atmosphere. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by preparative TLC (petroleum ether:ethyl acetate = 3:1) to afford the title compound (20.0 mg, 38% yield) as an orange oil. LCMS: m / z 572.3 [M+H] + ,ESI pos.

[0203] Step D: 3-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol To Pd / C (60.0 mg, purity: 10%) in 2 mL of EtOAc / methanol (0.20 mL) was added [(1R,2R)-2-[3-(2-benzyloxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl]cyclohexoxy]-tert-butyl-dimethyl-silane (20.0 mg, 0.03 mmol, 1.0 equiv.) and the mixture was stirred under H atmosphere at 25° C. for 1 h. The mixture was filtered through Celite, and the filtrate was concentrated in vacuo to give the title compound (15.0 mg, 26%) as an orange oil. LCMS: m / z 482.2 [M+H] + ,ESI pos.

[0204] Step E: 3-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol; 2,2,2-trifluoroacetic acid A solution of 3-[8-[(1R,2R)-2-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol (15.0 mg, 0.03 mmol, 1.0 equiv.) in HCl / dioxane (0.5 mL, 2 M in dioxane) was stirred at 25 °C for 0.08 h. The mixture was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Gemini 150 mm × 25 mm × 10 μm; mobile phase: [water (0.1% TFA, v / v)-MeCN]; B%: 12%–42%, 12 min) to give the title compound (3.7 mg, 24% yield) as a white solid. LCMS: m / z 368.2 [M+H] + ,ESI pos.

[0205] Example 16: 5-[8-(4-hydroxycyclohexyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol [ka] Step A: trans-4-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]cyclohexanamine A solution of 2-(3,6-dichloropyridazin-4-yl)oxyacetaldehyde (600.0 mg, 2.9 mmol, 1.0 equiv) and trans-4-[tert-butyl(dimethyl)silyl]oxycyclohexanamine (665.01 mg, 2.9 mmol, 1.0 equiv) in DCE (12 mL) was stirred at 25 °C for 10 min. Then, NaBH(OAc) (1225.97 mg, 5.8 mmol, 2.0 equiv) was added portionwise at 25 °C. The reaction mixture was stirred at 25 °C for 30 min. After the reaction was complete, the mixture was quenched with water (20 mL), then EtOAc (40 mL) was added, and the layers were separated. The aqueous phase was extracted with EtOAc (30 mL × 3). The combined extracts were washed with brine (30 mL × 2), dried over Na SO , filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (EA:MeOH=1:0 to 5:1) to give the title compound (490.0 mg, 40% yield) as a white solid. LCMS m / z: 420.2 [M+H] + ,ESI pos.

[0206] Step B: tert-Butyl-[4-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane To a mixture of trans-4-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]cyclohexanamine (490.0 mg, 1.17 mmol, 1.0 equiv.), CsCO (1.14 g, 3.5 mmol, 3.0 equiv.) in 1,4-dioxane (10 mL) under N was added Xphos Pd G (0.2 g, 0.23 mmol, 0.2 equiv.), and the mixture was stirred at 90 °C for 2 h. After the reaction was complete, the mixture was cooled to room temperature. EtOAc (20 mL) and water (10 mL) were then added, and the layers were separated. The aqueous phase was extracted with EtOAc (20 mL × 3). The combined extracts were washed with brine (10 mL × 2), dried over NaSO, filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (petroleum ether:ethyl acetate=1:2 to 2:1) to give the title compound (80.0 mg, 16% yield) as a yellow solid. LCMS m / z: 384.1 [M+H] + ,ESI pos.

[0207] The isomeric tert-butyl-[4-(3-chloro-6,7-dihydropyridazino[3,4-b][1,4]oxazin-5-yl)cyclohexoxy]-dimethyl-silane (60.0 mg, 9% yield) was obtained as a yellow solid.

[0208] Step C: 5-[8-[4-[tert-butyl(dimethyl)silyl]oxycyclohexyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol To a solution of tert-butyl-[4-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)-cyclohexoxy]-dimethyl-silane (70.0 mg, 0.18 mmol, 1.01 equiv.) and 6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (50.0 mg, 0.18 mmol, 1.0 equiv.) in 1,4-dioxane (1 mL) / water (0.2 mL) under N was added NaCO (57.57 mg, 0.54 mmol, 3.0 equiv.), and XPhos Pd g (30.69 mg, 0.04 mmol, 0.2 equiv.). The reaction mixture was heated under microwave irradiation at 95° C. for 2 hours under N2. After the reaction was complete, the reaction mixture was cooled to room temperature. Then, EtOAc (10 mL) and water (5 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (10 mL×3). The combined extracts were washed with brine (10 mL×2), dried over Na2SO4, filtered, concentrated in vacuo, and the crude was combined with another batch. The residue was analyzed by preparative TLC (PE / EA 4:1, R f =0.3) to give the title compound (35.0 mg, 34% yield) as a yellow solid. LCMS m / z: 498.3 [M+H] + ,ESI pos.

[0209] Step D: 5-[8-(4-hydroxycyclohexyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol To a solution of 5-[8-[4-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol (25.0 mg, 0.05 mmol, 1.0 equiv.) in 1,4-dioxane (0.500 mL) was added HCl / dioxane (0.05 mL, 0.05 mmol, 1.0 equiv.), and the mixture was stirred at 25° C. for 10 minutes. After the reaction was complete, the mixture was concentrated under vacuum. The residue was purified by HPLC. 18Purification by column chromatography (0.1% TFA / MeCN in water, MeCN: 20% to 30%, 4 min) gave LCMS m / z: 384.1 [M+H] + , ESI pos. gave the title compound.

[0210] Example 17: 5-[8-[(1R,3S)-3-Hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol [ka] Step A: (1R,3S)-3-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]-cyclohexanamine To a solution of 2-(3,6-dichloropyridazin-4-yl)oxyacetaldehyde (300.0 mg, 1.45 mmol, 1.0 equiv) in DCE (6 mL) was added (1R,3S)-3-[tert-butyl(dimethyl)silyl]oxycyclohexanamine (332.51 mg, 1.45 mmol, 1.0 equiv), followed by NaBH(OAc) (614.46 mg, 2.90 mmol, 2.0 equiv), and the mixture was stirred at 25° C. for 0.5 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (50 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 to give the crude product, which was purified by column chromatography on silica gel (petroleum ether:ethyl acetate=1:0 to 0:1) to give the title compound as a white solid (90 mg, 12% yield).

[0211] Step B: tert-Butyl-[(1R,3S)-3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane To a solution of (1R,3S)-3-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]cyclohexanamine (90.0 mg, 0.21 mmol, 1.0 equiv.) in 1,4-dioxane (3 mL) under N was added CsCO (208.7 mg, 0.64 mmol, 3.0 equiv.) and XphosPdG (36.84 mg, 0.04 mmol, 0.2 equiv.), and the reaction was stirred at 90 °C for 2 h under N. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (50 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 to give the crude product, which was combined with another batch. The combined crude products were purified by column chromatography on silica gel (petroleum ether:ethyl acetate=1:0 to 0:1) to give the title compound as a yellow oil (50.0 mg, 61% yield). LCMS m / z: 384.2 [M+H] + ,ESI pos. isomer tert-butyl-[(1S,3R)-3-(3-chloro-6,7-dihydropyridazino[3,4-b][1,4]oxazin-5-yl)cyclohexoxy]-dimethyl-silane (40.0 mg, 46% yield) was obtained as a yellow oil.

[0212] Step C: 5-[8-[(1R,3S)-3-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol To a solution of tert-butyl-[(1R,3S)-3-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)cyclohexoxy]-dimethyl-silane (40.0 mg, 0.1 mmol, 1.0 equiv) in 1,4-dioxane (2 mL) / water (0.200 mL) was added NaCO (22.08 mg, 0.21 mmol, 2.0 equiv), 6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (34.52 mg, 0.13 mmol, 1.2 equiv), and Xphos Pd g (17.66 mg, 0.02 mmol, 0.2 equiv). The reaction mixture was then stirred at 95° C. under N for 2 hours. The above reaction mixture was combined with another batch, diluted with water (20 mL), and extracted with ethyl acetate (50 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 to give the crude product. The crude product was analyzed by preparative TLC (petroleum ether:ethyl acetate=4:1, R f =0.4) to give the title compound (30.0 mg, 34% yield) as a yellow solid. LCMS m / z: 498.3 [M+H] + ,ESI pos.

[0213] Step D: 5-[8-[(1R,3S)-3-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol To a solution of [8-[(1R,3S)-3-[tert-butyl(dimethyl)silyl]oxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol (20.0 mg, 0.04 mmol, 1.0 equiv.) in 1,4-dioxane (2 mL), dioxane / HCl (1.0 mL, 2.0 mmol, 49.77 equiv.) was added, and the mixture was stirred at 25° C. for 0.08 h. The mixture was then concentrated in vacuo and combined with another batch. The pH was adjusted to approximately 8 with aqueous NaHCO (0.4 mL), then filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The residue was purified by preparative HPLC (column: Phenomenex Gemini, 150 mm x 25 mm x 5 μm; mobile phase: [water (0.1% NH H O, V / V)-ACN]; B%: 10%-40%, 10 min) to give the title compound (10.3 mg, 66% yield) as a white solid. LCMS m / z: 384.2 [M+H] + ,ESI pos.

[0214] Example 18: 5-[8-[(3S,4R)-3-hydroxytetrahydropyran-4-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol [ka] Step A: (3S,4R)-3-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]tetrahydropyran-4-amine To a solution of 2-(3,6-dichloropyridazin-4-yl)oxyacetaldehyde (320.0 mg, 1.55 mmol, 1.0 equiv.) in DCE (5 mL), (3S,4R)-3-[tert-butyl(dimethyl)silyl]oxytetrahydropyran-4-amine (429.26 mg, 1.85 mmol, 1.2 equiv.) was added, and the mixture was stirred at 20 °C for 5 minutes. Then, NaBH(OAc) (655.24 mg, 3.09 mmol, 2.0 equiv.) was added portionwise to the above mixture. The reaction mixture was then stirred at 20 °C for 5 hours. After the reaction was complete, the reaction mixture was cooled to room temperature. EtOAc (40 mL) and water (40 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (30 mL × 2). The combined extracts were washed with brine (60 mL), dried over NaSO, filtered, and concentrated in vacuo to give a residue. The residue was purified by column chromatography on silica gel (petroleum ether:ethyl acetate = 3:1 to 2:1) to give the title compound (400.0 mg, 55% yield) as a yellow oil. LCMS m / z: 422.2 [M+H] + ,ESI pos.

[0215] The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=3:1 to 2:1) to give the title compound (400.0 mg, yield 55%) as a yellow oil.

[0216] To a mixture of the above (3S,4R)-3-[tert-butyl(dimethyl)silyl]oxy-N-[2-(3,6-dichloropyridazin-4-yl)oxyethyl]tetrahydropyran-4-amine (350.0 mg, 0.83 mmol, 1.0 equiv.), CsCO (807.9 mg, 2.49 mmol, 3.0 equiv.) in 1,4-dioxane (20 mL) under N was added Xphos Pd G (142.58 mg, 0.17 mmol, 0.2 equiv.), followed by stirring at 90 °C for 2 h under N.

[0217] After the reaction was completed, the reaction mixture was cooled to room temperature. EtOAc (50 mL) and water (40 mL) were added, and the layers were separated. The aqueous phase was extracted with EtOAc (50 mL x 2). The combined extracts were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was purified by column chromatography on silica gel (petroleum ether:ethyl acetate = 5:1 to 3:1) to give the title compound (90.0 mg, 24% yield) as a yellow oil. LCMS m / z: 386.2 [M+H] + ,ESI pos.

[0218] Step C: 5-[8-[(3S,4R)-3-[tert-butyl(dimethyl)silyl]oxytetrahydropyran-4-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol To a solution of tert-butyl-[(3S,4R)-4-(3-chloro-6,7-dihydropyridazino[4,3-b][1,4]oxazin-8-yl)tetrahydropyran-3-yl]oxy-dimethyl-silane (60.0 mg, 0.13 mmol, 1.0 equiv.) in 1,4-dioxane (3 mL) and water (0.30 mL) was added NaCO (28.3 mg, 0.27 mmol, 2.0 equiv.), 6-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol (36.9 mg, 0.13 mmol, 1.0 equiv.) and Xphos Pd g3 (22.66 mg, 0.03 mmol, 0.2 equiv) was added and then stirred at 95° C. for 2 hours under N2. After the reaction was complete, the reaction mixture was cooled to room temperature. EtOAc (40 mL) and water (30 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (30 mL x 2). The combined extracts were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated in vacuo to give a residue. The residue was analyzed by preparative TLC (PE:EA=2.5:1, R f =0.2) to give the title compound (50.0 mg, 77% yield) as a yellow oil. LCMS m / z: 500.3 [M+H]+ ,ESI pos.

[0219] Step D: 5-[8-[(3S,4R)-3-hydroxytetrahydropyran-4-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol To a solution of 5-[8-[(3S,4R)-3-[tert-butyl(dimethyl)silyl]oxytetrahydropyran-4-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol (40.0 mg, 0.08 mmol, 1.0 equiv.) in 1,4-dioxane (3 mL), HCl / dioxane (2.0 mL, 2 M in water) was added. The reaction mixture was then stirred at 20° C. for 30 hours. The reaction mixture was concentrated under vacuum at 30° C. The residue was then dissolved in MeOH (3 mL), adjusted to pH 7 with NaHCO3, and then purified by reverse phase flash (CombiFlash, mobile phase: (water (0.1% ammonia hydroxide / v)-ACN); B%: 23%-25%) to give the title compound (20.3 mg, 64% yield) as a white solid. LCMS m / z: 386.1 [M+H] + ,ESI pos.

[0220] Example A The compound of formula I can be used in a manner known per se as the active ingredient to produce tablets of the following composition: 425mg per tablet Active ingredient 200mg Microcrystalline cellulose 155mg Cornstarch 25mg Talc 25mg Hydroxypropyl methylcellulose 20mg

[0221] Example B The compound of formula I can be used in a manner known per se as the active ingredient to produce capsules of the following composition: 220.0mg per capsule Active ingredient 100.0mg Cornstarch 20.0mg Lactose 95.0mg Talc 4.5mg Magnesium stearate 0.5mg

Claims

1. Formula I 【Chemistry 47】 [In the formula, R １ is H or alkyl; R ２ teeth, i. a 5-6 membered heterocycle containing a single O or a single N heteroatom, said heterocycle being optionally substituted with 1-2 substituents selected from alkyl, OH, halo, haloalkyl, hydroxyalkyl, or oxo; ii. a 5-6 membered heterocycle containing a single O heteroatom; iii. a 4- to 6-membered cycloalkyl optionally substituted with 1 to 2 substituents independently selected from alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl Selected from: X is —O—, —CH ２ -, -NH-, or -N(CH ３ )- and; W is a ring system A and B 【Chemistry 48】 【Chemistry 49】 (R ３ is H, halo, alkyl, alkoxy, or alkoxyalkyl, where R １ and R ３ one of which is H and the other is not H; R ４ is halo, alkyl, alkoxy, cyano, haloalkyl, or haloalkoxy; R ５ is H; or R ４ and R ５ , and the atoms to which they are attached, i. a 4- to 5-membered cycloalkyl ring optionally substituted with oxo, or ii. A 5-membered heterocycle containing a single O heteroatom optionally substituted with 1 to 2 substituents independently selected from alkyl and halo. Forming; R ６ is halo, haloalkyl, or OH; R ７ is H or F) Selected from and pharmaceutically acceptable salts thereof.

2. R １ is H or alkyl, and R ３ is H, alkyl or alkoxyalkyl, where R １ or R ３ 10. The compound of claim 1, wherein one of is H and the other is not H.

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

4. R ２ but, i. a 6-membered heterocycle containing a single N heteroatom substituted with alkyl; ii. -CH ２ -heterocycle, wherein the heterocycle is a 5-6 membered heterocycle containing a single O heteroatom, -CH ２ - heterocycles; iii. 4-membered cycloalkyl substituted with alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl The compound according to any one of claims 1 to 3, selected from:

5. R ２ but, 5-6 membered heterocycles containing a single O or a single N heteroatom, said heterocycles being optionally substituted by alkyl or by both alkyl and OH; and 4- to 6-membered cycloalkyl optionally substituted with 1 to 2 substituents independently selected from alkyl and OH The compound according to any one of claims 1 to 4, selected from:

6. X is O or —CH ２ The compound according to any one of claims 1 to 5, wherein

7. W is a ring system A, C and D [Transformation 50] 【Chemistry 51】 【Chemistry 52】 wherein Y is CH ２ or —O—. The compound according to any one of claims 1 to 6, selected from:

8. W is ring system A 【Chemistry 53】 The compound according to any one of claims 1 to 7,

9. R ４ The compound of any one of claims 1 to 8, wherein is alkyl, cyano, haloalkyl, or haloalkoxy.

10. R ４ The compound of any one of claims 1 to 9, wherein is cyano.

11. R ５ is H or R ４ and R ５ , and the atoms to which they are attached, i. a 4- to 5-membered cycloalkyl ring optionally substituted with oxo, or ii. 5-membered heterocycles containing a single O heteroatom The compound according to any one of claims 1 to 10, which forms

12. R ５ is H or R ４ and R ５ and the atoms to which they are attached form either a 5-membered cycloalkyl ring or a 5-membered heterocycle containing a single O heteroatom.

13. R ５ The compound according to any one of claims 1 to 11, wherein is H.

14. R ６ The compound according to any one of claims 1 to 12, wherein is OH.

15. R １ is H or alkyl; R ２ but, i. a 5-6 membered heterocycle containing a single O or a single N heteroatom, said heterocycle being optionally substituted with alkyl, halo, haloalkyl, hydroxyalkyl, or oxo; ii. -CH ２ -heterocycle, wherein the heterocycle is a 5-6 membered heterocycle containing a single O heteroatom, -CH ２ - heterocycles; iii. a 4- to 6-membered cycloalkyl optionally substituted with 1 to 2 substituents independently selected from alkyl and OH; and iv. 9-membered bicyclic heterocycles containing a single N heteroatom optionally substituted with alkyl Selected from: X is —O—, —CH ２ -, -NH-, or -N(CH ３ )- and; W is a ring system A and B 【Chemistry 54】 【Transformation 55】 (R ３ is H, halo, alkyl, alkoxy, or alkoxyalkyl, where R １ and R ３ one of which is H and the other is not H; R ４ is halo, alkyl, alkoxy, cyano, haloalkyl, or haloalkoxy; R ５ is H; or R ４ and R ５ , and the atoms to which they are attached, i. a 4- to 5-membered cycloalkyl ring optionally substituted with oxo, or ii. A 5-membered heterocycle containing a single O heteroatom optionally substituted with 1 to 2 substituents independently selected from alkyl and halo. Forming; R ６ is halo, haloalkyl, or OH; R ７ is H or F) Selected from: A compound according to claim 1; and pharmaceutically acceptable salts thereof.

16. R １ is H or alkyl, and R ３ is H or alkyl, where R １ or R ３ one of which is H and the other is alkyl; R ２ but, i. a 6-membered heterocycle containing a single N heteroatom substituted with alkyl, and ii. 4-membered cycloalkyl substituted with alkyl and OH Selected from: X is O or —CH ２ - and; W is a ring system A: 【Transformation 56】 and R ４ is cyano; R ５ is H; R ６ is OH, A compound according to any one of claims 1 to 13; and pharmaceutically acceptable salts thereof.

17. R １ is H or alkyl; R ２ but, i. 5-6 membered heterocycles containing a single O or a single N heteroatom, said heterocycles being optionally substituted with alkyl or with both alkyl and OH; ii. 4- to 6-membered cycloalkyl optionally substituted with 1 to 2 substituents independently selected from alkyl and OH Selected from: X is —O— or —CH ２ - and; W is a ring system A, C and D 【Chemistry 57】 【Transformation 58】 【Chemistry 59】 (In the formula, R ３ is H or alkyl, where R １ and R ３ one of which is H and the other is not H; R ４ is alkyl, cyano, haloalkyl, or haloalkoxy; R ５ is H; R ６ is OH; Y is CH ２ or O) Selected from: A compound according to claim 1; and pharmaceutically acceptable salts thereof.

18. R １ is H or alkyl; R ２ is a 5-6 membered heterocycle containing a single O or a single N heteroatom, said heterocycle being optionally substituted by alkyl or by both alkyl and OH; X is —O— or —CH ２ - and; W is a ring system C and D 【Transformation 60】 【Chemistry 61】 (In the formula, R ３ is H or alkyl, where R １ and R ３ one of which is H and the other is not H; R ４ is alkyl, cyano, haloalkyl, or haloalkoxy; R ５ is H; R ６ is OH; Y is CH ２ or O) Selected from: A compound according to claim 1; and pharmaceutically acceptable salts thereof.

19. 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; formic acid; 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 4-[8-[(3R)-1-ethyl-3-piperidyl]-6,7-dihydro-5H-pyrido[2,3-c]pyridazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 3-hydroxy-4-[8-(3-hydroxy-3-methyl-cyclobutyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; The compound of any one of claims 1 to 14, selected from: and pharmaceutically acceptable salts thereof.

20. 3-hydroxy-4-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid; 3-hydroxy-4-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-5-methyl-benzonitrile; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol; 2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol; 2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-2,3-dihydrobenzofuran-4-ol; 4-[8-[(3R,5S)-1-ethyl-5-hydroxy-3-piperidyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethyl)phenol; 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 2,2,2-trifluoroacetic acid; 4-[8-[(3R)-1-ethylpyrrolidin-3-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-5-methyl-benzonitrile; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol; 2,2,2-trifluoroacetic acid 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3,5-dimethyl-phenol; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol; 2,2,2-trifluoroacetic acid; 2-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-methyl-5-(trifluoromethoxy)phenol; 4-[8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-benzonitrile; 2,2,2-trifluoroacetic acid; 4-[8-[(3R)-1-ethyl-3-piperidyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-3-hydroxy-benzonitrile; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol; 2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]indan-4-ol; 3-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol; 2,2,2-trifluoroacetic acid; 3-[8-[(1R,2R)-2-hydroxycyclohexyl]-4-methyl-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]bicyclo[4.2.0]octa-1,3,5-trien-2-ol; 5-[8-[(1R,3S)-3-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; 5-[8-[(3S,4R)-3-hydroxytetrahydropyran-4-yl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; The compound of any one of claims 1 to 14, selected from: and pharmaceutically acceptable salts thereof.

21. 5-[8-(4-hydroxycyclohexyl)-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; The compound of any one of claims 1 to 14, selected from: and pharmaceutically acceptable salts thereof.

22. 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-2,3-dihydrobenzofuran-4-ol; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol; 2,2,2-trifluoroacetic acid; 5-[8-[(1R,2R)-2-hydroxycyclohexyl]-6,7-dihydropyridazino[4,3-b][1,4]oxazin-3-yl]-6-methyl-indan-4-ol; The compound of any one of claims 1 to 14, selected from: and pharmaceutically acceptable salts thereof.

23. A compound according to any one of claims 1 to 22 for use as a therapeutically active substance.

24. A compound according to any one of claims 1 to 22 for use in the treatment or prevention of a disease, disorder or condition responsive to NLRP3 inhibition.

25. A pharmaceutical composition comprising a compound according to any one of claims 1 to 22 and a therapeutically inert carrier.

26. 23. Use of a compound according to any one of claims 1 to 22 for the treatment or prevention of a disease, disorder or condition responsive to NLRP3 inhibition.

27. 23. A compound according to any one of claims 1 to 22 for use in the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

28. 23. Use of a compound according to any one of claims 1 to 22 in the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

29. 23. Use of a compound according to any one of claims 1 to 22 for the preparation of a medicament for the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

30. 23. A method of inhibiting NLRP3, comprising administering an effective amount of a compound according to any one of claims 1 to 22 to inhibit NLRP3.

31. 23. A method for the treatment or prevention of a disease, disorder or condition, said method comprising administering an effective amount of a compound according to any one of claims 1 to 22, wherein said disease, disorder or condition is selected from Alzheimer's disease and Parkinson's disease.

32. 10. The invention as hereinbefore described.

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