TRPV4 receptor ligands

Abstract

The present invention relates to TRPV4 receptor ligands. The present invention specifically describes receptor ligands for transient receptor potential cation channel subfamily V member 4 (TRPV4), pharmaceutical compositions comprising such compounds, and methods of using such compounds and compositions in the treatment of ocular disorders.

Description

[0001] This application is a divisional application of international application PCT / US2020 / 061587, which entered the Chinese national phase on May 20, 2022, with application number 202080080479.8 and invention name “TRPV4 receptor ligand”.

[0002] CROSS REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Patent Application No. 62 / 938,693, filed on November 21, 2019, which is incorporated herein by reference in its entirety. Technical Field

[0004] The present disclosure relates to compounds, compositions and methods for treating TRPV4-related diseases and / or disorders, such as glaucoma and related diseases. Background Art

[0005] Transient receptor potential cation channel subfamily V member 4 (TRPV4) is a member of the OSM9-like transient receptor potential channel (OTRPC) subfamily and is encoded by the TRPV4 gene in humans. TRPV4 protein is a Ca 2+ A permeability nonselective cation channel that is thought to be involved in the regulation of systemic osmotic pressure. TRPV4 also functions as a ciliary mechanosensory channel. Mutations in the TRPV4 gene have been associated with disorders including brachytruncus type 3, congenital distal spinal muscular atrophy, scapulohumeral spinal muscular atrophy, and Charcot-Marie-Tooth disease subtype 2C.

[0006] Defects in cilia formation or maintenance underlie a number of human diseases collectively referred to as ciliopathies, including retinitis pigmentosa, renal cysts, polydactyly, and developmental delay. OCRL, an inositol polyphosphate 5-phosphatase that regulates cilia biogenesis, has been linked to Lowe oculocerebrorenal syndrome (Lowe syndrome), a rare X-linked recessive disorder that manifests in males with bilateral cataracts and glaucoma as well as renal failure, hypotonia, and mental retardation. Substrates for OCRL include phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] and phosphatidylinositol-3,4,5-triphosphate [PI(3,4,5)P3]. Decreased 5-phosphatase activity and a 2- to 3-fold increase in the PI(4,5)P2:PI(4)P ratio have been demonstrated in fibroblasts from Lowe patients.

[0007] Mechanoreception of pressure underlies many important human diseases including the development of hypertension and glaucoma. In renal epithelial cells, the ciliary proteins polycystin (PC1 / 2) have been shown to be important for flow-dependent calcium flux. In the lining of the brain ventricles, cerebrospinal fluid is regulated by cilia. Similar to the kidney, the eye is a closed organ with sensitive homeostatic regulation of fluid production and outflow. Defective pressure sensing may cause an imbalance in the aqueous humor, leading to increased intraocular pressure. Low intraocular pressure can lead to structural changes in the retina and decreased visual acuity, whereas elevated intraocular pressure may damage the optic nerve. Glaucoma, an optic neuropathy associated with increased intraocular pressure, is the leading cause of irreversible blindness in the world.

[0008] Trabecular meshwork (TM) cells are responsible for the majority of the drainage of aqueous humor. Dysfunction of trabecular outflow leads to increased intraocular pressure, which in susceptible individuals causes death of retinal ganglion cells, leading to irreversible vision loss. The trabecular meshwork cells of the eye have primary cilia that are responsive to changes in pressure.

[0009] Consistent with the central role of increased pressure in the pathology of glaucoma, the only proven treatment is to reduce the pressure. There is a need to develop treatments for reducing intraocular pressure.

[0010] A hallmark of glaucoma is optic neuropathy in the posterior eye (retina), reflecting inflammatory activation of retinal glial cells and degeneration and loss of retinal ganglion cells (RGCs). The major risk factor for developing the disease is elevated intraocular pressure (IOP), with IOP dependence accounting for ~30–70% of glaucoma cases. Although “glaucoma” is a late name for the damaged retina, RGCs are extremely sensitive to acute changes in pressure, which lead to immediate changes in excitability, loss of temporal and spatial contrast sensitivity, and decreased visual acuity. Dysfunction is associated with retraction, thinning, and reduced complexity of RGC dendrites, loss of synapses, and changes in axonal transport. Pressure-induced changes in cellular firing and dendritic damage are exacerbated by impaired (anterograde / retrograde) transport of neurotrophic factors and organelles, which, if persistent, may lead to gross structural remodeling at the optic nerve head. A plausible multicompartmental regulator of pressure-induced damage may be calcium ions, which mediate the pathogenesis of optic neurite outgrowth through Ca ions. 2+ Dependent proteases, caspases, MAP kinases, ER stress, autophagy, and apoptosis drive remodeling of dendrites, cell bodies, and axons.

[0011] Currently recognized glaucoma treatments are primarily limited to lowering IOP. Because lowering IOP can also mitigate the progression of vision loss in patients with “normal” IOP (8-15 mm Hg) levels, glaucoma can be viewed as a disease involving pathological cellular mechanotransduction that increases the susceptibility of RGCs to mechanical stress at any IOP level (Krizaj, 2019). Unfortunately, the most commonly used IOP-lowering agent, latanoprost (a prostaglandin F2α analog), targets the secondary uveoscleral mechanism of aqueous humor drainage and does not reduce IOP by more than 20% in a significant portion (25-50%) of patients, while up to 6% of POAG patients do not respond to or are intolerant to any IOP medication. Therefore, an attractive strategy is to target both the IOP-generating mechanisms in the anterior eye and the pressure sensitivity of retinal ganglion cells and glia.

[0012] The present invention is based on the following discovery, that is, the pressure sensitivity of retinal ganglion cells, Muller glial cells and microglial cells is mediated in part by TRPV4 (transient receptor potential vanilloid subtype 4), which is a calcium permeable channel activated by a series of mechanical stressors (strain, pressure, compression, shear), moderate temperature and polyunsaturated fatty acids. Excessive activation of the channel induces selective death (Ryskamp et al., Journal of Neuroscience 2011) and reactive gliosis (Ryskamp et al., Journal of Neuroscience 2014) of RGCs, and inhibition or genetic ablation of the channel has a neuroprotective effect (Ryskamp et al., Scientific Reports 2016). It is believed that TRPV4 antagonists can be used to provide neuroprotection in ocular hypertension by inhibiting the pressure sensitivity of both glial cells (reducing neuroinflammation) and RGCs (inhibiting neurodegeneration). They are also believed to counteract the pressure-dependent calcium overload that may damage RGC dendrites and axons in glaucoma. Because these compounds also reduce IOP (Ryskamp et al., Scientific Reports 2016), the present invention provides a therapeutic strategy that combines IOP reduction with neuroprotection. In addition, the efficacy of TRPV4 antagonists in reducing glial and neuronal swelling and cytokine release in the retina (Jo et al., Journal of Neuroscience 2015; Matsumoto et al., Journal of Neuroscience 2018) provides a neuroprotective strategy for treating excessive swelling and / or edema in diseases such as diabetic retinopathy and macular degeneration.

[0013] Ryskamp DA, Jo AO, Frye A, Macaulay N, Vazquez-Chona F, Thoreson WB and D (2014). Swelling and eicosanoid metabolites differentially gate TRPV4 channels in retinal neurons and glia. Journal of Neuroscience 34(47):15689-15700. PMID: 25411497; PMC4236400.

[0014] Jo AO*, Ryskamp DA*, Phuong TTT, Verkman A, Yarishkin O, MacAulay N and D (2015). Synergistic signaling of TRPV4 and AQP4 channels is required for cell volume and calcium homeostasis regulation in retinal Müller glia. Journal of Neuroscience, 35(39): 13525-13537. PMID: 26424896. PMC4588615

[0015] Ryskamp DA, Frye AM, Phuong TTT, Yarishkin O, Jo AO, Xu Y, Lakk M, Redmon S, Iuso A, Hageman G, Ambati B, Prestwich GD, Torrejon KY and D (2016), TRPV4 regulates calcium homeostasis, cytoskeletal remodeling, conventional outflow and intraocular pressure in the mammalian eye, Scientific Reports 6:30583. PMID: 27510430. PMC4980693

[0016] Matsumoto H,Sugio S, D, Ishizaki Y and Shibasaki K (2018). Retinal detachment-induced Müller glial cell swelling activates TRPV4 ion channels and triggers rod photoreceptor death. Journal of Neuroscience 38(41):8745-8758. PMID: 30143574. PMC6181316

[0017] D (2019), What is glaucoma? Webvision: The Organization of the Retina and Visual System, edited by H Kolb, E Fernandez, and R Nelson, University of Utah Health Sciences, Salt Lake City. PMID 31241881 Summary of the invention

[0018] In one aspect, disclosed are compounds of formula (I):

[0019]

[0020] or a pharmaceutically acceptable salt thereof, wherein:

[0021] L 1 is -C(O)-, -S(O2)-, -S(O)- or C1-C4 alkylene;

[0022] X is O or NR 1 ;

[0023] m is 0, 1, 2, 3, or 4;

[0024] n is 0, 1, 2, 3, or 4;

[0025] Ar 1 and Ar 2 Each is independently selected from a 5-10 membered aryl or heteroaryl group;

[0026] Z is for CHR 2 R 3 , COR 4 or NHR 5 ;

[0027] R 1 is selected from hydrogen and C1-C4 alkyl;

[0028] R 2 , R 3 , R 4 and R 5 Each is independently selected from hydrogen, hydroxy, C1-C6 alkyl, hydroxy-C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C1-C6 alkyl, phosphate, C1-C6 alkyl phosphate, C4-C8 heterocyclyl, NR 6 R 7 、-OC(O)R 8 、-C(O)R 9 、-S(O)2R 10 and -OS(O)2R 11 ;

[0029] R 6 , R 7 , R 8 , R 9 , R 10 and R 11 each independently selected from hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C4-C8 cycloalkyl, C4-C8 heterocyclyl, C5-C8 aryl, C5-C8 heteroaryl, C1-C6 alkoxy-C1-C6 alkyl, C5-C8 aryl-C1-C6 alkyl, C5-C8 heteroaryl-C1-C6 alkyl, C5-C8 heterocyclyl-C1-C6 alkyl, amino-C1-C6 alkyl, hydroxy-C1-C6 alkyl, hydroxy-C3-C8 cycloalkyl, cyano-C1-C6 alkyl, hydroxysulfonyl-C1-C6 alkyl, phosphate-C1-C6 alkyl, alkylphosphate-C1-C6 alkyl, -COR 12 and-CR 13 R 14 C(O)R15 ;

[0030] R 12 Selected from C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, C4-C8 heterocyclyl, hydroxy-C1-C6 alkyl, amino-C4-C8 heterocyclyl, phosphate-C1-C6 alkyl and C1-C6 alkyl phosphate;

[0031] R 13 and R 14 are each independently selected from hydrogen, C1-C6 alkyl and C1-C6 heteroalkyl, or optionally taken together with the atoms to which they are attached to form a ring; and

[0032] R 15 is hydroxy, C1-C6 alkoxy or amino;

[0033] wherein at least one of m and n is not zero, and each aryl, heteroaryl, cycloalkyl or heterocycle is independently unsubstituted or substituted with one or more substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, haloalkyl, sulfonyl, -C(O)-C1-C6 alkoxy and -C(O)-NH2.

[0034] Also disclosed are pharmaceutical compositions comprising the compounds, and methods of using the compounds and compositions to treat disorders such as ocular hypertension or glaucoma or other related TRPV4-related medical disorders and / or diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 Graph showing the effect of two concentrations of topical Compound 11 (0.01% and 0.1%), 0.5% timolol, and control PBS on IOP in the microbead (MB) mouse model of ocular hypertension (N=3 independent experiments with 7-10 mice per experimental group). Error bars=±SEM. Ocular hypertension was induced using intracameral injection of polystyrene microbeads in a mouse model of primary open-angle glaucoma (POAG). Antagonist stock solutions (10 mM) were prepared in DMSO and final concentrations were prepared in PBS and diluted to <0.0001% DMSO.

[0036] Figure 2is a bar graph showing the cumulative data of the IOP pre-elevation group of mouse eyes (N≥3 independent experiments, with 7-10 mice per experimental group). Left panel: IOP data are plotted as % of control values ​​for the timolol treated eye group. Middle panel: MB injections raise IOP in all experimental groups. Right panel: Topical application of timolol, compound 11 and compound 52 and intraperitoneal injection of HC067047 significantly reduced IOP levels in MB treated eyes (**P<0.01; ****P<0.001; two-way ANOVA followed by Holm- test). DETAILED DESCRIPTION

[0037] Disclosed herein are receptor ligands (e.g., antagonists) of transient receptor potential cation channel subfamily V member 4 (TRPV4). The compound may have a structure of formula (I). The compound of formula (I) may exhibit selectivity for TRPV4 over other TRPV receptors (e.g., TRPV3). By modulating TRPV4 activity, the compound of formula (I) may be used to treat or prevent diseases and disorders associated with TRPV4. For example, the compound of formula (I) is used to treat or prevent ocular diseases and disorders such as glaucoma.

[0038] 1. Definition

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art. In the event of a conflict, the present document, including the definitions, shall prevail. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All publications, patent applications, and other references mentioned herein are incorporated herein by reference in their entirety. The materials, methods, and examples disclosed herein are merely illustrative and are not intended to be limiting.

[0040] When used herein, the terms "include", "comprising", "having", "may", "containing" and variations thereof are intended to be open transitional phrases, terms or words that do not exclude the possibility of other arrangements or configurations. Unless the context clearly states otherwise, references without a specific number include plural referents.

[0041] The modifier "about" used in conjunction with a quantity includes the stated value and has the meaning dictated by the context (e.g., it includes at least the degree of error associated with the measurement of the particular quantity in question). The modifier "about" should also be considered to disclose a range defined by the absolute values ​​of the two endpoints. For example, the expression "about 2 to about 4" also discloses a range of "2 to 4". The term "about" can refer to plus or minus 10% of the specified number. For example, "about 10%" can mean a range of 9% to 11%, and "about 1" can mean 0.9-1.1. Other meanings of "about" may be apparent from the context, such as rounding, so that, for example, "about 1" may also mean 0.5 to 1.4.

[0042] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics 75th edition, inside cover), and specific functional groups are generally defined as described therein. In addition, general principles of organic chemistry as well as specific functional components and reactivities are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999, Smith and March, March's Advanced Organic Chemistry, 5th edition, John Wiley & Sons, Inc., New York, 2001, Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989, Carruthers, Some Modern Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987, the entire contents of each of which are incorporated herein by reference.

[0043] The term "alkoxy" as used herein refers to an alkyl group, as defined herein, attached to the parent molecular moiety through an oxygen atom. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and tert-butoxy.

[0044] As used herein, the term "alkyl" means a straight or branched saturated hydrocarbon chain containing 1 to 10 carbon atoms. The term "short chain alkyl" or "C 1- "C6 alkyl" means a straight or branched hydrocarbon containing 1 to 6 carbon atoms. The term "C1-C3 alkyl" means a straight or branched hydrocarbon containing 1 to 3 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl.

[0045] The term "alkoxyalkyl" as used herein, means an alkoxy group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.

[0046] The term "alkylamino" as used herein means at least one alkyl group, as defined herein, appended to the parent molecular moiety through an amino group, as defined herein.

[0047] As used herein, the term "alkyl phosphate" means an ester of phosphoric acid H3PO4 in which any or all of the hydrogens may be replaced by an alkyl group as defined herein, such that the alkyl phosphate may be a monosubstituted R 1 H2PO4, disubstituted R 1 R 2 H2PO4 or trisubstituted R 1 R 2 R 3 H2PO4, where R 1 , R 2 and R 3 The alkyl groups may be the same or different.

[0048] The term "alkylphosphate alkyl" as used herein, refers to an alkyl phosphate, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein.

[0049] As used herein, the term "amide" means -C(O)NR- or -NRC(O)-, where R can be hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, heterocycle, alkenyl, or heteroalkyl.

[0050] As used herein, the term "amino" means -NR x R y , where R x and R ycan be hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, heterocyclic, alkenyl or heteroalkyl. In the case of aminoalkyl or any other moiety in which the amino group is linked to two other moieties, the amino group can be –NR x –, where R x It may be hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, heterocycle, alkenyl or heteroalkyl.

[0051] The term "aminoalkyl" as used herein means at least one amino group, as defined herein, appended to the parent molecular moiety through an alkylene group, as defined herein.

[0052] The term "aminoheterocyclyl" as used herein means at least one amino group, as defined herein, appended to the parent molecular moiety through a heterocyclyl group, as defined herein.

[0053] As used herein, the term "aryl" refers to phenyl or a bicyclic fused ring system. Examples of bicyclic fused ring systems are phenyl attached to a parent molecular moiety and fused to a cycloalkyl as defined herein, a phenyl, a heteroaryl as defined herein, or a heterocycle as defined herein. Representative examples of aryl include, but are not limited to, indolyl, naphthyl, phenyl, quinolyl, and tetrahydroquinolyl.

[0054] The term "arylalkyl" as used herein means at least one aryl group, as defined herein, appended to the parent molecular moiety through an alkylene group, as defined herein.

[0055] As used herein, the term "cyano" means at least one -CN group.

[0056] The term "cyanoalkyl" as used herein means at least one -CN group attached to the parent molecular moiety through an alkylene group as defined herein.

[0057] As used herein, the term "cycloalkyl" refers to a carbocyclic ring system containing 3 to 10 carbon atoms, zero heteroatoms, and zero double bonds. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecyl. "Cycloalkyl" also includes carbocyclic ring systems in which the cycloalkyl is attached to a parent molecular moiety and fused to an aryl as defined herein (e.g., phenyl), a heteroaryl as defined herein, or a heterocycle as defined herein. Representative examples of such cycloalkyl groups include, but are not limited to, 2,3-dihydro-1H-indenyl (e.g., 2,3-dihydro-1H-inden-1-yl and 2,3-dihydro-1H-inden-2-yl), 6,7-dihydro-5H-cyclopenta[b]pyridinyl (e.g., 6,7-dihydro-5H-cyclopenta[b]pyridin-6-yl), and 5,6,7,8-tetrahydroquinolinyl (e.g., 5,6,7,8-tetrahydroquinolin-5-yl).

[0058] As used herein, the term "fluoroalkyl" means an alkyl group as defined herein in which 1, 2, 3, 4, 5, 6, 7 or 8 hydrogen atoms are replaced by fluorine. Representative examples of fluoroalkyl groups include, but are not limited to, 2-fluoroethyl, 2,2,2-trifluoroethyl, trifluoromethyl, difluoromethyl, pentafluoroethyl and trifluoropropyl groups such as 3,3,3-trifluoropropyl.

[0059] As used herein, the term "halogen" or "halo" means Cl, Br, I, or F.

[0060] The term "haloalkyl" as used herein means an alkyl group as defined herein wherein 1, 2, 3, 4, 5, 6, 7 or 8 hydrogen atoms are replaced by halogen.

[0061] As used herein, the term "heteroalkyl" means an alkyl group as defined herein wherein one or more carbon atoms have been replaced by a heteroatom selected from S, O, P and N. Representative examples of heteroalkyl groups include, but are not limited to, alkyl ethers, secondary and tertiary alkyl amines, amides, and alkyl thioethers.

[0062] As used herein, the term "heteroaryl" refers to an aromatic monocyclic or aromatic bicyclic ring system. The aromatic monocyclic ring is a 5- or 6-membered ring containing at least one heteroatom independently selected from N, O and S (e.g., 1, 2, 3 or 4 heteroatoms independently selected from O, S and N). The 5-membered aromatic monocyclic ring has two double bonds, and the 6-membered aromatic monocyclic ring has three double bonds. Examples of the bicyclic heteroaryl are monocyclic heteroaromatic rings attached to a parent molecular component and fused to a monocyclic cycloalkyl as defined herein, a monocyclic aryl as defined herein, a monocyclic heteroaryl as defined herein, or a monocyclic heterocycle as defined herein. Representative examples of heteroaryl include, but are not limited to, indolyl, pyridyl (including pyridin-2-yl, pyridin-3-yl, pyridin-4-yl), pyrimidinyl, pyrazinyl, pyridazinyl, pyrazolyl, pyrrolyl, benzopyrazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4- Oxazolyl, 1,2,4- oxadiazole, imidazolyl, thiazolyl, isothiazolyl, thienyl, benzimidazolyl, benzothiazolyl, benzo Azolyl, benzo oxadiazolyl, benzothiophenyl, benzofuranyl, isobenzofuranyl, furanyl, Azolyl, iso oxazolyl, purinyl, isoindolyl, quinoxalinyl, indazolyl, quinazolinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, isoquinolyl, quinolyl, 6,7-dihydro-1,3-benzothiazolyl, imidazo[1,2-a]pyridinyl, naphthyridinyl, pyridoimidazolyl, thiazo[5,4-b]pyridin-2-yl, thiazo[5,4-d]pyrimidin-2-yl.

[0063] The term "heteroarylalkyl" as used herein means at least one heteroaryl group, as defined herein, appended to the parent molecular moiety through an alkylene group, as defined herein.

[0064] As used herein, the term "heterocycle" or "heterocyclic" means a monocyclic heterocycle, a bicyclic heterocycle or a tricyclic heterocycle. The monocyclic heterocycle is a 3, 4, 5, 6, 7 or 8 membered ring containing at least one heteroatom independently selected from O, N and S. The 3 or 4 membered ring contains zero or one double bond and a heteroatom selected from O, N and S. The 5 membered ring contains zero or one double bond and 1, 2 or 3 heteroatoms selected from O, N and S. The 6 membered ring contains 0, 1 or 2 double bonds and 1, 2 or 3 heteroatoms selected from O, N and S. The 7 and 8 membered rings contain 0, 1, 2 or 3 double bonds and 1, 2 or 3 heteroatoms selected from O, N and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridine, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiocyclopentanyl, 1,3-dithiocyclohexanyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidin ... Oxazoline, iso Oxazolidinyl, morpholinyl, 2-keto-3-piperidinyl, 2-ketoazetidin-3-yl, Diazoline, Oxazolidinyl, Oxazoline, oxazolidinyl, oxetanyl, oxepanyl, oxocanyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydrothiophenyl, thiadiazolinyl, thiadiazolidinyl, 1,2-thiazinyl, 1,3-thiazinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, and trithianyl. The bicyclic heterocycle is a monocyclic heterocycle fused to a phenyl, or a monocyclic heterocycle fused to a monocyclic cycloalkyl, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl, or a monocyclic heterocycle fused to a monocyclic heterocycle, or a spiroheterocyclyl, or a bridged monocyclic heterocyclic ring system in which two non-adjacent atoms of the ring are connected by an alkylene bridge of 1, 2, 3 or 4 carbon atoms or an alkenylene bridge of 2, 3 or 4 carbon atoms. Representative examples of bicyclic heterocycles include, but are not limited to, benzopyranyl, benzothiopyranyl, chromanyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzothiophenyl, 2,3-dihydroisoquinolinyl, 2-azaspiro[3.3]hept-2-yl, 2-oxa-6-azaspiro[3.3]hept-6-yl, azabicyclo[2.2.1]heptyl (including 2-azabicyclo[2.2.1]hept-2-yl), azabicyclo[3.1.0]hexyl (including 3-azabicyclo[3.1.0]hex-3-yl), 2,3-dihydro-1H-indolyl, isoindolyl, octahydrocyclopenta[c]pyrrolyl, octahydropyrrolopyridinyl, and tetrahydroisoquinolinyl. Examples of tricyclic heterocycles are bicyclic heterocycles fused to phenyl, or bicyclic heterocycles fused to monocyclic cycloalkyl, or bicyclic heterocycles fused to monocyclic cycloalkenyl, or bicyclic heterocycles fused to monocyclic heterocycles, or bicyclic heterocycles wherein two non-adjacent atoms of the bicyclic ring are linked by an alkylene bridge of 1, 2, 3, or 4 carbon atoms, or an alkenylene bridge of 2, 3, or 4 carbon atoms. Examples of tricyclic heterocycles include, but are not limited to, octahydro-2,5-epoxypentalene, hexahydro-2H-2,5-methylene-bridged cyclopenta[b]furan, hexahydro-1H-1,4-methylene-bridged cyclopenta[c]furan, aza-adamantane (1-azatricyclo[3.3.1.13,7]decane), and oxa-adamantane (2-oxatricyclo[3.3.1.13,7]decane). The monocyclic, bicyclic and tricyclic heterocycles are attached to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the rings and may be unsubstituted or substituted.

[0065] The term "heterocyclylalkyl" as used herein means at least one heterocycle, as defined herein, appended to the parent molecular moiety through an alkylene group, as defined herein.

[0066] As used herein, the term "hydroxy" means an -OH group.

[0067] The term "hydroxyalkyl" as used herein means at least one -OH group attached to the parent molecular moiety through an alkylene group as defined herein.

[0068] The term "hydroxycycloalkyl" as used herein means at least one -OH group is attached to the parent molecular moiety through a cycloalkyl group as defined herein.

[0069] Hydroxysulfonyl

[0070] The term "hydroxysulfonyl-alkyl" as used herein means at least one hydroxysulfonyl group, as defined herein, appended to the parent molecular moiety through an alkylene group, as defined herein.

[0071] The term "phosphate alkyl" as used herein means at least one phosphate group attached to the parent molecular moiety through an alkylene group as defined herein.

[0072] As used herein, the term "sulfonyl" means a -SO2- group.

[0073] In some cases, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkyl or cycloalkyl) is indicated by the prefix "C x -C y -" where x is the minimum number of carbon atoms in the substituent and y is the maximum number. Thus, for example, "C1-C3 alkyl" refers to an alkyl substituent containing from 1 to 3 carbon atoms.

[0074] The term "substituent" refers to a group "substituted" on an aryl, heteroaryl, phenyl or pyridyl group at any atom of the group. Any atom may be substituted.

[0075] The term "substituted" means that a group can be further substituted by one or more non-hydrogen substituents. Substituents include, but are not limited to, halogen, =O (keto), =S (thioketo), cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, ketone, amide, carbamate, and acyl. For example, if a group is described as "optionally substituted" (e.g., alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heteroalkyl, heterocycle, or other groups such as R groups), it may have 0, 1, 2, 3, 4, or 5 substituents independently selected from halogen, =0 (keto), =S (thioketo), cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, keto, amide, carbamate, and acyl.

[0076] the term refers to a single bond or double bond

[0077] For the compounds described herein, groups and substituents thereof may be chosen based on the valences allowed by atoms and substituents such that the selection and substitution results in stable compounds, eg, compounds that do not spontaneously undergo transformations by rearrangement, cyclization, elimination, and the like.

[0078] For the recitation of numerical ranges herein, each intervening number with the same precision is expressly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range of 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9 and 7.0 are expressly contemplated.

[0079] 2. Compounds

[0080] In one aspect, disclosed are compounds of formula (I):

[0081]

[0082] or a pharmaceutically acceptable salt thereof, wherein:

[0083] L 1 is -C(O)-, -S(O2)-, -S(O)- or C1-C4 alkylene;

[0084] X is O or NR 1 ;

[0085] m is 0, 1, 2, 3, or 4;

[0086] n is 0, 1, 2, 3, or 4;

[0087] Ar 1 and Ar 2 Each is independently selected from a 5-10 membered aryl or heteroaryl group;

[0088] Z is for CHR 2 R 3 , COR 4 or NHR 5 ;

[0089] R 1 is selected from hydrogen and C1-C4 alkyl;

[0090] R 2 , R 3 , R 4 and R 5 Each is independently selected from hydrogen, hydroxy, C1-C6 alkyl, hydroxy-C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C1-C6 alkyl, phosphate, C1-C6 alkyl phosphate, C4-C8 heterocyclyl, NR 6 R 7 、-OC(O)R 8 、-C(O)R 9 、-S(O)2R 10 and -OS(O)2R 11 ;

[0091] R 6 , R 7 , R 8 , R 9 , R 10 and R 11each independently selected from hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C4-C8 cycloalkyl, C4-C8 heterocyclyl, C5-C8 aryl, C5-C8 heteroaryl, C1-C6 alkoxy-C1-C6 alkyl, C5-C8 aryl-C1-C6 alkyl, C5-C8 heteroaryl-C1-C6 alkyl, C5-C8 heterocyclyl-C1-C6 alkyl, amino-C1-C6 alkyl, hydroxy-C1-C6 alkyl, hydroxy-C3-C8 cycloalkyl, cyano-C1-C6 alkyl, hydroxysulfonyl-C1-C6 alkyl, phosphate-C1-C6 alkyl, alkylphosphate-C1-C6 alkyl, -COR 12 and-CR 13 R 14 C(O)R 15 ;

[0092] R 12 Selected from C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, C4-C8 heterocyclyl, hydroxy-C1-C6 alkyl, amino-C4-C8 heterocyclyl, phosphate-C1-C6 alkyl and C1-C6 alkyl phosphate;

[0093] R 13 and R 14 are each independently selected from hydrogen, C1-C6 alkyl and C1-C6 heteroalkyl, or optionally taken together with the atoms to which they are attached to form a ring; and

[0094] R 15 is hydroxy, C1-C6 alkoxy or amino;

[0095] wherein at least one of m and n is not zero, and each aryl, heteroaryl, cycloalkyl or heterocycle is independently unsubstituted or substituted with one or more substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, haloalkyl, sulfonyl, -C(O)-C1-C6 alkoxy and -C(O)-NH2.

[0096] In certain embodiments, L 1 It is -S(O2)-.

[0097] In certain embodiments, Ar 1 Selected from 5-membered aryl, 5-membered heteroaryl, 6-membered aryl or 6-membered heteroaryl. Each aryl or heteroaryl can be unsubstituted or substituted by one or two substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy, haloalkyl, sulfonyl and-C(O)-C1-C6 alkoxy. In some embodiments, the halogen is fluorine or chlorine. In some embodiments, the haloalkyl is trifluoromethyl.

[0098] In certain embodiments, Ar 1 yes

[0099]

[0100] In an exemplary embodiment, Ar 1 yes

[0101] In certain embodiments, Ar 2 Selected from 5-membered aryl, 5-membered heteroaryl, 6-membered aryl or 6-membered heteroaryl. Each aryl or heteroaryl can be unsubstituted or substituted by one or two substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy,-C(O)-C1-C6 alkoxy and-C(O)-NH2. In some embodiments, the halogen is fluorine or chlorine.

[0102] In certain embodiments, Ar 2 yes

[0103] In an exemplary embodiment, Ar 2 yes

[0104] In certain embodiments, Z is CHR 2 R 3 , COR 4 or NHR 5 ;

[0105] R 2 and R 3 Each independently selected from hydrogen, hydroxyl, C1-C6 alkoxy, C4-C8 heterocyclic group, phosphate, NR 6 R 7 、-OC(O)R 8 and S(O)2R 10 ; R 4 Selected from hydroxyl, C1-C6 alkoxy, and NR 6 R 7 ;

[0106] R 5 Selected from hydrogen, C1-C6 alkyl, -C(O)R 9 and -S(O)2R 10 ;

[0107] R 6 and R 7Each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C4-C8 heterocyclyl, C1-C6 alkoxy-C1-C6 alkyl, C5-C8 heteroaryl-C1-C6 alkyl, C5-C8 heterocyclyl-C1-C6 alkyl, amino-C1-C6 alkyl, hydroxy-C1-C6 alkyl, hydroxy-C3-C8 cycloalkyl, cyano-C1-C6 alkyl, hydroxysulfonyl-C1-C6 alkyl, alkyl phosphate-C1-C6 alkyl, -COR 12 and -CHR 13 C(O)R 14 ;

[0108] R 8 is C5-C8 aryl or C5-C8 heteroaryl;

[0109] R 9 Selected from C1-C6 alkyl, C1-C6 heteroalkyl, amino-C1-C6 alkyl, hydroxy-C1-C6 alkyl or -COR 12 ;

[0110] R 10 is amino-C1-C6 alkyl;

[0111] R 12 Selected from C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, C4-C8 heterocyclyl, amino-C4-C8 heterocyclyl, phosphate-C1-C6 alkyl and C1-C6 alkyl phosphate;

[0112] R 13 and R 14 are each independently selected from hydrogen and C1-C6 alkyl; and

[0113] R 15 Selected from hydroxy, C1-C6 alkoxy and amino.

[0114] In exemplary embodiments, Z is -CH2OH, -C(O)OH, NH2, -NHC(O)CH2, -CH2NH2, -CH2NHCH3, -CH2NH(CH2)2OH, -CH2NH(CH2)3OH, -CH2NH(C5H9O), -CH2NH(CH2)2OCH3, -CH2NHCH2(C4H7O), -CH2NHCH2CH3, -CH2NHCH2CH(OH)CH3, -CH2NH(C3H5O), or -CH2NHCH2(C3H5O).

[0115] In certain embodiments, X is O and n is 0. In certain embodiments, m is 1. In certain embodiments, X is O, m is 1, and n is 0.

[0116] In certain embodiments, X is NR 1 In certain embodiments, R 1 It's hydrogen.

[0117] In certain embodiments, X is NR 1 and m is 0. In certain embodiments, X is NR 1 , m is 0, and n is 1.

[0118] In certain embodiments, X is NR 1 and n is 0. In certain embodiments, X is NR 1 , m is 1, and n is 0.

[0119] A compound selected from:

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130] The compounds may exist as stereoisomers in which asymmetric or chiral centers are present. Depending on the configuration of the substituents around the chiral carbon atom, the stereoisomers are "R" or "S". The terms "R" and "S" used herein are configurations defined in the IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem., 1976, 45: 13-30. Various stereoisomers and mixtures thereof are contemplated by the present disclosure, and they are specifically included within the scope of the present disclosure. Stereoisomers include enantiomers and diastereomers and mixtures of enantiomers or diastereomers. Each stereoisomer of the compound can be synthesized from commercially available starting materials containing asymmetric or chiral centers, or by preparing a racemic mixture and then using a resolution method known to those of ordinary skill in the art. Examples of these resolution methods are (1) attachment of a mixture of enantiomers to a chiral auxiliary, separation of the resulting diastereomeric mixture by recrystallization or chromatography, and optionally liberation of the optically pure product from the auxiliary, as described in Furniss, Hannaford, Smith and Tatchell, Vogel's Textbook of Practical Organic Chemistry, 5th edition (1989), Longman Scientific & Technical, Essex CM202 JE, England), or (2) direct separation of the mixture of optical enantiomers on a chiral chromatographic column, or (3) fractional recrystallization methods.

[0131] It is to be understood that the compounds may have tautomeric forms as well as geometric isomers and these also constitute embodiments of the present disclosure.

[0132] The present disclosure also includes isotopically labeled compounds, which are the same as those described in formula (I), but one or more atoms are replaced by atoms with an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes suitable for inclusion in the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as but not limited to 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35S. 18 F and 36 Cl. Using heavier isotopes such as deuterium (i.e. 2 H) may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in certain circumstances. The compounds may incorporate positron emitting isotopes for use in medical imaging and positron emission tomography (PET) studies to determine receptor distribution. Suitable positron emitting isotopes that may be incorporated into compounds of formula (I) are 11 C. 13 N. 15 O and 18 F. Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples, using an appropriate isotopically-labelled reagent in place of a non-isotopically-labelled reagent.

[0133] In certain embodiments, in the compounds of Formula (I), any hydrogen atom may be deuterium.

[0134] The disclosed compounds can act or be used as antagonists of TRPV4. In certain embodiments, the disclosed compounds can be used with an IC in the range of about 0.1 nM to about 50 μM, about 0.1 nM to about 10 μM, about 0.1 nM to about 1 μM, about 0.1 nM to about 100 nM, or about 0.1 nM to about 50 nM. 50Inhibits TRPV4. In certain embodiments, the disclosed compounds can be less than 50 μM, less than 49 μM, less than 48 μM, less than 47 μM, less than 46 μM, less than 45 μM, less than 44 μM, less than 43 μM, less than 42 μM, less than 41 μM, less than 40 μM, less than 39 μM, less than 38 μM, less than 37 μM, less than 36 μM, less than 35 μM, less than 34 μM, less than 33 μM, less than 32 μM, less than 31 μM, less than 30 μM, less than 29 μM, less than 28 μM, less than 27 μM, less than 26 μM, less than 25 μM, less than 24 μM, less than 23 μM, less than 22 μM, less than 21 μM, less than 20 μM, less than 19 μM, less than 18 μM, less than 17 μM, less than 16 μM, less than 18 μM, less than 19 μM, less than 18 ... An IC of less than 15 μM, less than 14 μM, less than 13 μM, less than 12 μM, less than 11 μM, less than 10 μM, less than 9 μM, less than 8 μM, less than 7 μM, less than 6 μM, less than 5 μM, less than 4 μM, less than 3 μM, less than 2 μM, less than 1 μM, less than 950 nM, less than 900 nM, less than 850 nM, less than 800 nM, less than 750 nM, less than 700 nM, less than 650 nM, less than 600 nM, less than 550 nM, less than 500 nM, less than 450 nM, less than 400 nM, less than 350 nM, less than 300 nM, less than 250 nM, less than 200 nM, less than 150 nM, less than 100 nM, less than 50 nM, less than 10 nM, less than 5 nM or less than 1 nM. 50 Inhibits TRPV4.

[0135] The disclosed compounds may be selective modulators of TRPV4 relative to other TRPV receptors such as TRPV3.

[0136] The disclosed compounds may exist as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to salts or zwitterions of the compound that are soluble or dispersible in water or oil, suitable for treating disorders without excessive toxicity, irritation and allergic reactions, commensurate with a reasonable benefit / risk ratio, and effective for its intended use. The salts may be prepared during the final separation and purification of the compound, or separately prepared by reacting the amino group of the compound with a suitable acid. For example, the compound may be dissolved in a suitable solvent such as, but not limited to, methanol and water, and treated with at least one equivalent of an acid such as hydrochloric acid. The resulting salt may be precipitated and separated by filtration and dried under reduced pressure. Alternatively, the solvent and excess acid may be removed under reduced pressure to provide a salt. Representative salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthylsulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, p-toluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, phosphate, etc. The amino groups of the compounds may also be quaternized with chlorinated, brominated and iodinated alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, etc.

[0137] Base addition salts can be prepared during the final separation and purification of the disclosed compounds by reaction of the carboxyl group with a suitable base, such as a metal cation such as a hydroxide, carbonate or bicarbonate of lithium, sodium, potassium, calcium, magnesium or aluminum, or an organic primary, secondary or tertiary amine. Quaternary ammonium salts can be prepared, for example, quaternary ammonium salts derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, benzhydrylamine, N,N-dibenzylphenethylamine, 1-diphenylhydroxymethylamine (1-ephenamine) and N,N'-dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, etc.

[0138] The compounds and intermediates can be separated and purified by methods known to professionals in the field of organic synthesis. Examples of conventional methods for separating and purifying compounds may include, but are not limited to, chromatography on solid supports such as silica gel, aluminum oxide, or silica derived from alkylsilane groups, by recrystallization at high or low temperatures and optionally pre-treatment with activated carbon, thin layer chromatography, distillation at various pressures, sublimation and grinding under vacuum, as described in Vogel's Textbook of Practical Organic Chemistry, 5th edition (1989), Furniss, Hannaford, Smith and Tatchell, Longman Scientific & Technical, Essex CM202JE, England).

[0139] The disclosed compounds may have at least one basic nitrogen, whereby the compounds may be treated with an acid to form the desired salt. For example, the compound may be reacted with an acid at or above room temperature to provide the desired salt, which is deposited after cooling and collected by filtration. Examples of acids suitable for the reaction include, but are not limited to, tartaric acid, lactic acid, succinic acid, and mandelic acid, atrolactic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, benzenesulfonic acid, carbonic acid, fumaric acid, maleic acid, gluconic acid, acetic acid, propionic acid, salicylic acid, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, citric acid, hydroxybutyric acid, camphorsulfonic acid, malic acid, phenylacetic acid, aspartic acid or glutamic acid, etc.

[0140] The reaction conditions and reaction times of each individual step can be changed according to the substituents present in the specific reactants used and the reactants used.Specific procedures are provided in the embodiment section.Reaction can be carried out in a conventional manner, for example, by removing the solvent from the residue and further purifying according to methods generally known in the art, such as but not limited to crystallization, distillation, extraction, grinding and chromatography.Unless otherwise indicated, starting material and reagent are commercially available or can be prepared from commercially available materials by the method described in the chemical literature by those skilled in the art.If starting material is not commercially available, it can be prepared by the program selected from standard organic chemical techniques, the technology similar to the synthesis of known structurally similar compounds or the technology similar to the program described in the above scheme or synthesis example part.

[0141] Routine experiments, including appropriate manipulation of reaction conditions, reagents and synthetic route sequences, protection of any chemical functional groups that are not compatible with the reaction conditions, and deprotection at a suitable point in the reaction sequence of the method, are all included within the scope of the present invention. Suitable protecting groups and methods for protecting and deprotecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples thereof can be found in Wuts and TW Greene, Greene's book entitled "Protective Groups in Organic Synthesis, (4th Edition), John Wiley & Sons, NY (2006), which is incorporated herein by reference as a whole. The synthesis of the compounds of the present invention can be accomplished by methods similar to those described in the synthesis schemes and specific embodiments described above.

[0142] When an optically active form of a disclosed compound is required, it can be obtained by carrying out one of the procedures described herein using optically active starting materials (e.g., preparation by asymmetric induction of an appropriate reaction step), or by resolving a mixture of stereoisomers of the compounds or intermediates using standard procedures (e.g., chromatographic separation, recrystallization, or enzymatic resolution).

[0143] Likewise, when a pure geometric isomer of a compound is required, it may be obtained by carrying out one of the above procedures using the pure geometric isomer as starting material, or by resolving a mixture of geometric isomers of a compound or intermediate using standard procedures such as chromatographic separation.

[0144] It can be appreciated that the synthetic schemes and specific examples described are illustrative and should not be construed as limiting the scope of the invention, as the scope of the invention is defined in the claims. All alternatives, modifications and equivalents of the synthetic methods and specific examples are included within the scope of the claims.

[0145] 3. Pharmaceutical Composition

[0146] The disclosed compounds can be incorporated into pharmaceutical compositions suitable for administration to a subject (eg, a patient, who can be human or non-human).

[0147] The pharmaceutical composition may include a "therapeutically effective amount" or a "prophylactically effective amount" of the agent. A "therapeutically effective amount" refers to an amount that is effective to achieve the desired therapeutic outcome at the required dosage and length of time. The therapeutically effective amount of a composition can be determined by one skilled in the art, and may vary with factors such as the disease state, age, sex, and weight of the individual, and the ability of the composition to elicit a desired response in the individual. A therapeutically effective amount is also an amount in which any toxic or deleterious effects of a compound of the invention (e.g., a compound of Formula (I)) are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount that is effective to achieve the desired prophylactic outcome at the required dosage and length of time. Typically, a prophylactic effective amount will be lower than a therapeutically effective amount because a prophylactic dose is used in a subject prior to or at an early stage of the disease.

[0148] For example, a therapeutically effective amount of a compound of formula (I) can be from about 1 mg / kg to about 1000 mg / kg, from about 5 mg / kg to about 950 mg / kg, from about 10 mg / kg to about 900 mg / kg, from about 15 mg / kg to about 850 mg / kg, from about 20 mg / kg to about 800 mg / kg, from about 25 mg / kg to about 750 mg / kg, from about 30 mg / kg to about 700 mg / kg, from about 35 mg / kg to about 650 mg / kg, from about 40 mg / kg to about 600 mg / kg, About 45 mg / kg to about 550 mg / kg, about 50 mg / kg to about 500 mg / kg, about 55 mg / kg to about 450 mg / kg, about 60 mg / kg to about 400 mg / kg, about 65 mg / kg to about 350 mg / kg, about 70 mg / kg to about 300 mg / kg, about 75 mg / kg to about 250 mg / kg, about 80 mg / kg to about 200 mg / kg, about 85 mg / kg to about 150 mg / kg and about 90 mg / kg to about 100 mg / kg.

[0149] The pharmaceutical composition may comprise a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars such as, but not limited to, lactose, glucose and sucrose, starches such as, but not limited to, corn starch and potato starch, cellulose and its derivatives such as, but not limited to, sodium carboxymethylcellulose, ethylcellulose and cellulose acetate, tragacanth powder, malt, gelatin, talc, excipients such as, but not limited to, cocoa butter and suppository waxes, oils such as, but not limited to, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol, esters such as, but not limited to, ethyl oleate and ethyl laurate, agar, buffers such as, but not limited to, magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, phosphate buffer, and other nontoxic compatible lubricants such as, but not limited to, sodium lauryl sulfate and magnesium stearate, and colorants, release agents, coating agents, sweeteners, flavoring and perfuming agents, preservatives and antioxidants may also be present in the composition according to the judgment of the formulator.

[0150] Thus, the compounds and their physiologically acceptable salts can be formulated for administration by, for example, solid administration, eye drops, topical oil-based formulations, injection, inhalation (by mouth or nose), implants, or oral, buccal, parenteral, or rectal administration. Techniques and formulations can generally be found in Remington's Pharmaceutical Sciences (Meade Publishing Co., Easton, Pa.). Therapeutic compositions must generally be sterile and stable under the conditions of manufacture and storage.

[0151] The route of administration of the disclosed compound and the form of the composition will determine the type of carrier used. The composition can take a variety of different forms, such as those suitable for systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implant or parenteral) or topical administration (e.g., skin, lung, nose, ear, eye, liposome delivery system or iontophoresis).

[0152] Carriers for systemic administration generally include at least one of a diluent, a lubricant, a binder, a disintegrant, a colorant, a flavoring agent, a sweetener, an antioxidant, a preservative, a glidant, a solvent, a suspending agent, a wetting agent, a surfactant, a combination thereof, etc. All carriers are optional in the composition.

[0153] Suitable diluents include sugars such as glucose, lactose, dextrose and sucrose, glycols such as propylene glycol, calcium carbonate, sodium carbonate, sugar alcohols such as glycerol, mannitol and sorbitol. The amount of diluent in the systemic or topical composition is generally from about 50 to about 90%.

[0154] Suitable lubricants include silicon dioxide, talc, stearic acid and its magnesium and calcium salts, calcium sulfate, and liquid lubricants such as polyethylene glycols and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter. The amount of lubricant in systemic or topical compositions is generally about 5 to about 10%.

[0155] Suitable binders include polyvinyl pyrrolidone, magnesium aluminum silicate, starches such as corn starch and potato starch, gelatin, tragacanth and cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, methylcellulose, microcrystalline cellulose and sodium carboxymethylcellulose. The amount of binder in the system composition is generally from about 5 to about 50%.

[0156] Suitable disintegrants include agar, alginic acid and its sodium salt, effervescent mixtures, cross-linked carboxymethylcellulose, cross-linked povidone, sodium carboxymethyl starch, sodium starch glycolate, clays and ion exchange resins. The amount of disintegrant in systemic or topical compositions is generally from about 0.1 to about 10%.

[0157] Suitable coloring agents include colorants such as FD&C dyes. When used, colorants are generally present in systemic or topical compositions in amounts of from about 0.005 to about 0.1%.

[0158] Suitable flavoring agents include menthol, peppermint, and fruit flavors.When used, flavoring agents are generally present in systemic or topical compositions in an amount of from about 0.1 to about 1.0%.

[0159] Suitable sweeteners include aspartame and saccharin.The amount of sweetener in the systemic or topical composition is generally from about 0.001 to about 1%.

[0160] Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of antioxidant in the systemic or topical composition is generally from about 0.1 to about 5%.

[0161] Suitable preservatives include benzalkonium chloride, methylparaben and sodium benzoate.The amount of preservative in systemic or topical compositions is generally from about 0.01 to about 5%.

[0162] Suitable glidants include silicon dioxide. The amount of glidant in the systemic or topical composition is generally from about 1 to about 5%.

[0163] Suitable solvents include water, isotonic saline, ethyl oleate, glycerol, hydroxylated castor oil, alcohols such as ethanol, and phosphate buffered saline.The amount of solvent in a systemic or topical composition is generally from about 0 to about 100%.

[0164] Suitable suspending agents include AVICEL RC-591 (available from FMC Corporation, Philadelphia, PA) and sodium alginate. The amount of suspending agent in systemic or topical compositions is generally from about 1 to about 8%.

[0165] Suitable surfactants include lecithin, polysorbate 80, sodium lauryl sulfate and TWEENS (from Atlas Powder Company, Wilmington, Delaware). Suitable surfactants include those disclosed in CTFA Cosmetic Ingredient Handbook (Cosmetic Ingredient Handbook, 1992, pp. 587-592), Remington's Pharmaceutical Sciences (Remington's Pharmaceutical Sciences, 15th Edition, 1975, pp. 335-337) and McCutcheon's Emulsifiers & Detergents, Volume 1, 1994, North American Edition, pp. 236-239). The amount of surfactant in the system or topical composition is generally from about 0.1% to about 5%.

[0166] Although the amounts of the components in the systemic composition may vary depending on the type of systemic composition prepared, in general, the systemic composition comprises 0.01% to 50% of the active compound (e.g., a compound of formula (I)) and 50% to 99.99% of one or more carriers. Compositions for parenteral administration typically comprise 0.1% to 10% of the active ingredient and 90% to 99.9% of carriers, including diluents and solvents.

[0167] Compositions for oral administration can have a variety of different dosage forms. For example, solid forms include tablets, capsules, granules and bulk powders. These oral dosage forms include a safe and effective amount, usually at least about 5%, more particularly about 25% to about 50% of the active ingredient. Oral dosage compositions contain about 50% to about 95%, more particularly about 50% to about 75% of a carrier.

[0168] Tablets can be compressed, tablet ground, enteric coated, sugar coated, film coated or compressed multiple times. Tablets generally include active ingredients and carriers, and the carrier includes ingredients selected from diluents, lubricants, binders, disintegrants, colorants, flavorings, sweeteners, glidants and combinations thereof. Specific diluents include calcium carbonate, sodium carbonate, mannitol, lactose and cellulose. Specific binders include starch, gelatin and sucrose. Specific disintegrants include alginic acid and cross-linked carboxymethyl cellulose. Specific lubricants include magnesium stearate, stearic acid and talc. Specific coloring agents are FD&C dyes, which can be added for appearance. Chewable tablets preferably contain sweeteners such as aspartame and saccharin or flavorings such as menthol, peppermint, fruit flavorings, or combinations thereof.

[0169] Capsules (including implants, time-release and sustained-release formulations) typically contain an active compound (e.g., a compound of formula (I)) and a carrier, the carrier comprising one or more diluents disclosed above, in a capsule comprising gelatin. Granules typically contain a disclosed compound and preferably contain a glidant such as silicon dioxide to improve flow characteristics. Implants may be biodegradable or non-biodegradable types.

[0170] The choice of ingredients in the carrier for oral compositions depends on secondary considerations such as taste, cost and storage stability and is not critical for the purposes of the present invention.

[0171] Solid compositions can be coated by conventional methods, typically with a pH or time-dependent coating, so that the disclosed compounds are released into the gastrointestinal tract near the desired site of administration or released at different sites and times to prolong the desired effect. The coating typically comprises a cellulose acetate phthalate, polyvinyl acetate phthalate, vinyl acetate, cellulose phthalate, hydroxypropyl methylcellulose phthalate, ethyl cellulose, One or more components of a coating (available from Evonik Industries, Essen, Germany), wax and shellac.

[0172] Compositions for oral administration can have liquid form.For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstructed by non-effervescent granules, suspensions reconstructed by non-effervescent granules, effervescent preparations reconstructed by effervescent granules, elixirs, tinctures, syrups, etc. The liquid oral administration composition generally includes a disclosed compound and a carrier, i.e., a carrier selected from diluents, colorants, flavorings, sweeteners, preservatives, solvents, suspending agents and surfactants. Oral liquid compositions preferably include one or more components selected from colorants, flavorings and sweeteners.

[0173] Other compositions that can be used to achieve systemic delivery of the subject compound include sublingual, buccal and nasal dosage forms. Such compositions generally include one or more soluble filler materials, such as diluents including sucrose, sorbitol and mannitol, and adhesives such as gum arabic, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methylcellulose. Such compositions may also include lubricants, colorants, flavorings, sweeteners, antioxidants and glidants.

[0174] The disclosed compounds can be administered topically. Topical compositions that can be topically applied to the skin can take any form, including solids, solutions, oils, creams, ointments, gels, lotions, shampoos, leave-on and rinse-off conditioners, emulsions, cleansers, moisturizers, sprays, skin patches, etc. Topical compositions include the disclosed compounds (e.g., compounds of formula (I)) and a carrier. The carrier of the topical composition preferably helps the compound penetrate into the skin. The carrier may also include one or more optional components.

[0175] The amount of carrier used in conjunction with the disclosed compounds is sufficient to provide a practical amount of the composition for administration per unit dose of the compound. Techniques and compositions for preparing dosage forms useful in the methods of the present invention are described in the following references: Modern Pharmaceutics, Chapters 9 and 10, Banker & Rhodes, ed. (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd ed., (1976).

[0176] The carrier may include a single component or a combination of two or more components. In topical compositions, the carrier includes a topical carrier. Suitable topical carriers include one or more components selected from phosphate buffer, isotonic water, deionized water, monofunctional alcohols, symmetrical alcohols, aloe vera gel, allantoin, glycerol, vitamin A and E oil, mineral oil, propylene glycol, PPG-2 myristyl propionate, dimethyl isosorbide, castor oil, combinations thereof, etc. More specifically, carriers for skin application include propylene glycol, dimethyl isosorbide and water, and even more specifically phosphate buffer, isotonic water, deionized water, monofunctional alcohols and symmetrical alcohols.

[0177] The carrier of the topical composition may also include one or more ingredients selected from the group consisting of emollients, propellants, solvents, humectants, thickeners, powders, fragrances, pigments and preservatives, all of which are optional.

[0178] Suitable emollients include stearyl alcohol, glyceryl monoricinoleate, glyceryl monostearate, propane-1,2-diol, butane-1,3-diol, mink oil, cetyl alcohol, isopropyl isostearate, stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecane-2-ol, isocetyl alcohol, cetyl palmitate, di-n-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, peanut oil, castor oil, acetylated lanolin alcohol, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, and combinations thereof. Specific emollients for use on the skin include stearyl alcohol and dimethicone.The amount of emollient in skin-based topical compositions is generally from about 5% to about 95%.

[0179] Suitable propellants include propane, butane, isobutane, dimethyl ether, carbon dioxide, nitrous oxide, and combinations thereof.The amount of propellant in the topical composition is generally from about 0% to about 95%.

[0180] Suitable solvents include water, ethanol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulfoxide, dimethylformamide, tetrahydrofuran and combinations thereof. Specific solvents include ethanol and isopropyl alcohol. The amount of solvent in the topical composition is generally from about 0% to about 95%.

[0181] Suitable humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, gelatin, and combinations thereof. Specific humectants include glycerin. The amount of humectant in the topical composition is typically 0% to 95%.

[0182] The amount of thickener in the topical composition is generally from about 0% to about 95%.

[0183] Suitable powders include beta-cyclodextrin, hydroxypropyl cyclodextrin, chalk, talc, Fuller's earth, kaolin, starch, gum, colloidal silicon dioxide, sodium polyacrylate, tetraalkyl ammonium montmorillonite, trialkyl aryl ammonium montmorillonite, chemically modified magnesium aluminum silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethylcellulose, ethylene glycol monostearate, and combinations thereof. The amount of powder in the topical composition is typically 0% to 95%.

[0184] The amount of fragrance in topical compositions is generally from about 0% to about 0.5%, especially from about 0.001% to about 0.1%.

[0185] Suitable pH adjusting additives include HCl or NaOH in an amount sufficient to adjust the pH of the topical pharmaceutical composition.

[0186] 4. Treatment Methods

[0187] The disclosed compounds can be used in methods for treating TRPV4-related medical disorders and / or diseases. The treatment methods may include administering to a subject in need of such treatment a composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0188] The compounds can be administered to a subject in need thereof to modulate TRPV4 for a variety of biological processes.

[0189] The compounds can be used to treat and prevent certain diseases and disorders associated with TRPV4 dysfunction in humans and animals. Treatment or prevention of such diseases and disorders can be achieved by modulating TRPV4 in a subject by administering a compound or composition of the present disclosure alone or in combination with another active agent as part of a treatment regimen to a subject in need thereof.

[0190] In combination therapy, the other drugs can be administered simultaneously or sequentially with the disclosed compounds by commonly used routes and amounts. When the disclosed compounds are used simultaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such drugs and the disclosed compounds is preferred. However, the combination therapy can also be administered according to overlapping schedules. It is also contemplated that the combination of one or more active ingredients with the disclosed compounds can be more effective than using any single agent.

[0191] The compounds can be used to treat or prevent diseases or disorders associated with TPRV4 dysfunction, such as eye diseases, such as retinopathy including non-proliferative and proliferative diabetic retinopathy and retinopathy of prematurity, glaucoma, macular degeneration, age-related macular degeneration (wet and dry), retinitis pigmentosa, Stargardt's disease, macular edema, uveitis, ocular hypertension, and retinal infections including infections caused by cytomegalovirus. The diseases may be chronic or acute. The compounds and pharmaceutical compositions disclosed herein can also be used to prevent blast eye injuries mediated by, for example, IEDs, high centrifugal forces during flight, and mechanical trauma. In one embodiment, the compounds and pharmaceutical compositions disclosed herein can be used to treat or prevent glaucoma.

[0192] The vertebrate retina, which is exposed to systemic blood pressure, hydrostatic pressure from the CSF, and intrinsic IOP, contains one or more pressure-sensitive TRPs and / or piezoelectric channels. Pathological elevations of IOP or systemic pressure are major risk factors for many conditions, such as glaucoma, a group of inherited optic neuropathies characterized by apoptotic loss of RGCs, optic nerve degeneration, and progressive loss of visual field. The cellular pathophysiology of glaucoma is not well understood, in part because the mechanisms coupling mechanical stimulation (ΔIOP) to cellular signaling remain to be characterized.

[0193] The compounds and compositions disclosed herein can be used to treat or prevent diseases and disorders other than ocular diseases for which TRPV4 channel antagonists may be beneficial. For example, compounds of formula (I) can be used to treat and / or prevent bladder disorders, heart failure, pulmonary edema, atherosclerosis, disorders associated with intestinal edema, postoperative abdominal edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone-related dysfunction and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator-induced lung injury, high altitude induced pulmonary edema, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, sinusitis / rhinitis, asthma, overactive bladder, pain, motor neuron disorders, inherited gain of function disorders, cardiovascular disease, renal insufficiency, osteoarthritis, Crohn's disease, colitis, diarrhea, intestinal irregularities (hyperresponsiveness / hyporesponsiveness), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance or flatulence.

[0194] Also provided is a method of treating one or more disorders in a subject for which TRPV4 receptor modulation is expected to be beneficial, the method comprising the step of administering to the subject at least one disclosed compound, at least one disclosed pharmaceutical composition, and / or at least one disclosed product in a dose and amount effective to treat the disorder in the subject.

[0195] The present disclosure relates to the use of the described chemical compositions for treating diseases or disorders for which TRPV4 receptor modulation is expected to have a therapeutic effect, such as atherosclerosis, disorders related to intestinal edema, fluid retention, sepsis, hypertension, inflammation, bone-related dysfunction and congestive heart failure, pulmonary disorders, overactive bladder, pain, motor neuron disorders, inherited gain-of-function disorders, cardiovascular disease, renal insufficiency, osteoarthritis, intestinal irregularities (hyper-responsiveness / hypo-responsiveness), in patients (preferably humans) by administering one or more of the disclosed compounds or products.

[0196] Also provided is a method of treating a disorder in a mammal, comprising the step of administering to the mammal at least one disclosed compound, composition, or medicament.

[0197] The compounds can further be used in methods for preventing, treating, controlling, ameliorating or reducing the risk of the diseases, disorders and conditions mentioned herein. The compounds can be further combined with other agents for use in methods for preventing, treating, controlling, ameliorating or reducing the risk of the diseases, disorders and conditions mentioned herein.

[0198] In addition, various ocular traumas can produce long-term degenerative effects on the eye. For example, subjects in close proximity to an explosion may develop subsequent degenerative conditions due to the compressive effects of the explosion on the eye. In some cases, this damage may not occur immediately, but rather over time. Treatment of such ocular trauma may include delivering a TRPV4 antagonist to the eye following such trauma to mitigate, reduce, or eliminate the associated long-term effects.

[0199] In addition, in some cases, apoptosis of RGCs may be associated with eye conditions that have a degenerative component. In some cases, TRPV4 receptor antagonists may have a neuroprotective effect on at least RGCs, thereby treating or alleviating the destructive effects of such conditions. It is noteworthy that treatment of the eye with a TRPV4 antagonist may be beneficial for subjects who experience an increase in IOP as well as those who do not show an increase in IOP or a moderate increase in IOP.

[0200] On the one hand, TRPV4 antagonists can target stress-induced Ca2+ in vitro and in vivo. 2+ This antagonist lowers IOP in the anterior chamber of the eye, suggesting that it regulates fluid production / absorption in the anterior trabecular meshwork and blocks pressure-induced RGC apoptosis.

[0201] TRPV4 ion channels are known to be involved in regulating calcium flux and are associated with retinal remodeling during chronic increases in IOP (Ryskamp et al., J. Neuroscience 2011, 31(19), 7089-7101, incorporated herein). In certain embodiments, the compounds of formula (I) can antagonize excessive TRPV4 activation. Regulating IOP by regulating fluid production in the anterior eye can be used to treat eye diseases, particularly glaucoma.

[0202] 5. Medicine box

[0203] In one aspect, the present disclosure provides a kit comprising at least one disclosed compound or a pharmaceutically acceptable salt thereof, and optionally one or more of the following:

[0204] (a) at least one agent known to modulate TRPV4 activity;

[0205] (b) at least one agent known to treat a disorder associated with TPRV4 activity;

[0206] (c) instructions for treating a disorder associated with TPRV4 activity; or

[0207] (d) instructions for administering the compound in conjunction with an ocular treatment.

[0208] In some embodiments, the at least one disclosed compound and at least one medicament are co-formulated. In some embodiments, the at least one disclosed compound and at least one medicament are co-packaged. The kit may also include compounds and / or products co-packaged, co-formulated, and / or co-delivered with other components. For example, a drug manufacturer, drug distributor, physician, pharmacy, or pharmacist may provide a kit comprising a disclosed compound and / or product and another component for delivery to a patient.

[0209] The disclosed kits can be used in conjunction with the disclosed methods of use.

[0210] The kit may contain information, instructions, or both, indicating that the use of the kit provides treatment for a medical condition in a mammal, particularly a human. The information and instructions may take the form of text, pictures, or both. Additionally or alternatively, the kit may include a compound, composition, or both, and information, instructions, or both regarding a method of administering the compound or composition, preferably useful for treating or preventing a medical condition in a mammal, such as a human.

[0211] The compounds and methods of the present invention will be better understood by reference to the following examples, which are intended as illustrations of the scope of the invention and not as limitations.

[0212] 6. Examples

[0213] The following examples are provided as illustrations of the partial scope and specific embodiments of the present disclosure and are not meant to limit the scope of the present disclosure. Unless otherwise indicated, abbreviations and chemical symbols have their usual and customary meanings. Unless otherwise indicated, the compounds described herein have been prepared, isolated and characterized using the schemes and other methods disclosed herein, or can be prepared using the described reaction routes and methods.

[0214] The preparation of starting materials that are commercially available, described in the literature, or readily available to one skilled in the art is not described. The skilled artisan will appreciate that where compounds are stated to be prepared analogously to earlier examples or intermediates, reaction times, equivalents of reagents, and temperatures may be modified for each specific reaction, and that different work-up or purification techniques may be necessary or desirable.

[0215] The following abbreviations may be used: lithium bis(trimethylsilyl)amide (LHMDS), diisopropyl azodicarboxylate (DIAD), trifluoroacetic acid (TFA), dichloromethane (DCM), triethylamine (Et3N), N,N-diisopropylethylamine (DIPEA), ethyl acetate (EtOAc), triphenylphosphine (PPh3), azabenzotriazole tetramethyluronium hexafluorophosphate (HATU), ethanol (EtOH), lithium tri-tert-butoxyaluminum hydride (LiAlH(Ot-Bu)3), borane dimethyl sulfide complex (BH3· SMe2), N,N-dimethylaminopyridine (DMAP), acetonitrile (CH3CN), diethyl ether (Et2O), tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), N,N'-dimethylformamide (DMF), sodium triacetoxyborohydride (NaBH(OAc)3), acetic acid (AcOH), 1,2-dichloroethane (DCE), sodium acetate (NaOAc), sodium cyanoborohydride (NaBH3CN), tert-butyldimethylsilyl chloride (TBS-Cl), and tetrabutylammonium fluoride (TBAF).

[0216] General Methods. All reactions requiring anhydrous conditions were performed in oven- or flame-dried glassware. Commercially available reagents were used as received; in other cases, materials were purified accordingly following the laboratory chemical purification procedures. Dichloromethane, N,N'-dimethylformamide, toluene, acetonitrile, and tetrahydrofuran were degassed with nitrogen and passed through a solvent purification system (Innovative Technologies Pure Solv). Anhydrous 1,4-dichloromethane was used as received. Alkane was purchased from Acros Organics and used in AcrosSeal TM In the bottle. Triethylamine and N,N-diisopropylethylamine Store on molecular sieves or in The reaction was distilled over 100 molecular sieves. Microwave reactions were performed in a 908005 CEM Discover System. The reaction was monitored by TLC, visualized by dual short / long wave UV lamps, and stained with KMnO4 or 12-phosphomolybdic acid in ethanol. Flash chromatography was performed on Merck silica gel Kieselgel 60 (230-400 mesh) from EMScience using specified HPLC grade solvents or using an automated medium pressure column chromatography system (Teledyne ISCO CombiFlash RF75 or CombiFlash Rf+). Reverse phase HPLC was performed on a Waters HPLC Semi Prep 150B system using a Sunfire C18 Prep column or an Atlantis T3 Prep column. LCMS data were analyzed on a Thermo ScientificTM UltiMate TM 3000 UHPLC, fluorescence detector monitored at 214 or 254nm. LC / MS / MS data were collected on a ThermoFinnigan TSQ Quantum with a Genesis Lightn C18, 4uM 50mm column (ID 2.1mm) in positive or negative mode. The mobile phase consisted of acetonitrile (0.1% formic acid) and water (0.1% formic acid) for positive ion mode and methanol (5mM ammonium bicarbonate) and water (5mM ammonium bicarbonate) for negative ion mode. 1 H NMR spectra were recorded at 500 MHz, 400 MHz and 300 MHz, 13C at 125 MHz. Proton resonances are reported relative to the deuterated solvent peak: 7.27 ppm for CDCl3, 3.31 ppm for CD3OD (centerline signal), 2.50 for DMSO-d6, 4.79 for D2O, using the following format: Chemical shift (δ) [multiplicity (s singlet, br s broad singlet, d doublet, t triplet, q quartet, m multiplet)]. Carbon resonances are reported as chemical shift (δ) in parts per million, relative to the centerline signal of the corresponding solvent peak: 77.23 ppm for CDCl3, 49.15 ppm for CD3OD.

[0217] Example 1 - Compound 1

[0218]

[0219] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-methylazetidin-3-yl)methoxy)benzonitrile was prepared according to Reaction Scheme 1:

[0220]

[0221] Reagents: Step 1) LiHMDS, CH3I, THF, –78°C to rt; 2) LiBH4, THF, CH3OH; 3) DIAD, PPh3, 4-hydroxybenzonitrile, THF; 4) TFA, DCM; 5) 2,4-dichlorobenzenesulfonyl chloride, Et3N, DCM.

[0222] Step 1.3-methylazetidine-1,3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester. In a 3-necked flask, LHMDS solution (1.0M, in THF, 11.3mL) was added to anhydrous THF (30mL) under N2, and the mixture was cooled to -78 ° C. A solution of azetidine-1,3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (2.00g, 9.29mmol) in anhydrous THF (20mL) was added dropwise, and the mixture was stirred at -78 ° C for 20min. A solution of iodomethane (0.75mL, 12.05mmol) in anhydrous THF (8mL) was added dropwise, and the mixture was warmed to rt and stirred for 3 days. The reaction was quenched with half-saturated NH4Cl (200mL) and extracted with EtOAc (200mL). The organic layer was washed with brine (200 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by flash chromatography (hexanes / EtOAc) to give the title compound as a light yellow oil (1.07 g). 1 HNMR (500MHz, CDCl3) δppm 4.21 (d, 2H), 3.75 (s, 3H), 3.67 (d, 2H), 1.53 (s, 3H), 1.44 (s, 9H).

[0223] Step 2. tert-butyl 3-(hydroxymethyl)-3-methylazetidine-1-carboxylate. To a solution of 1-(tert-butyl) 3-methyl azetidine-1,3-dicarboxylate (1.07 g, 4.67 mmol) in anhydrous THF (40 mL) was added LiBH4 (208 mg, 9.54 mmol), and the mixture was stirred at rt under N2 for 19 h. CH3OH (10 mL) was added to the reaction mixture along with another portion of LiBH4 (510 mg, 23.39 mmol), and stirring was continued at rt for 25 h. A third portion of LiBH4 (510 mg, 23.39 mmol) was added, and the mixture was stirred for another 25 h. The reaction was quenched with half-saturated NH4Cl (100 mL) and extracted with EtOAc (2×50 mL). The extract was washed with brine (75 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexanes / EtOAc) to give the title compound as a colorless semisolid (800 mg). 1 H NMR (500MHz, CDCl3) δppm 3.76 (d, 2H), 3.56-3.62 (m, 2H), 3.54 (d, 2H), 2.00-2.05 (m, 1H), 1.43 (s, 9H), 1.25 (s, 3H).

[0224] Step 3. tert-Butyl 3-((4-cyanophenoxy)methyl)-3-methylazetidine-1-carboxylate. To a solution of tert-butyl 3-(hydroxymethyl)-3-methylazetidine-1-carboxylate (103 mg, 0.512 mmol), 4-hydroxybenzonitrile (70 mg, 0.588 mmol) and PPh3 (230 mg, 0.877 mmol) in anhydrous THF (5 mL) was added neat DIAD (0.17 mL, 0.86 mmol) dropwise, and the mixture was stirred at rt and N2 for 18 h. The reaction mixture was diluted with EtOAc (30 mL), washed with saturated NaHCO3 aqueous solution (2×25 mL) and brine (25 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (234 mg) with a purity of about 60%.

[0225] Step 4.4-((3-methylazetidine-3-yl)methoxy)benzonitrile. A mixture of tert-butyl 3-((4-cyanophenoxy)methyl)-3-methylazetidine-1-carboxylate (purity of about 60%, 234 mg, 0.46 mmol) and TFA (1.5 mL) in anhydrous DCM (7 mL) was stirred at rt under N2 for 24 h. The reaction mixture was concentrated in vacuo and the residue was partitioned between 1M HCl (15 mL) and DCM (15 mL). The aqueous layer was washed with another portion of DCM (15 mL), adjusted to alkaline (about pH 12) with 1M NaOH (20 mL), and extracted with DCM (3×15 mL). The extract was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound (60 mg) as a colorless oil. 1 H NMR (500MHz, CDCl3) δppm7.54-7.63(m,2H),6.95-7.01(m,2H),4.03(s,2H),3.58(d,2H),3.45(d,2H),2.07(br s,1H),1.38(s,3H).

[0226] Step 5. 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-methylazetidin-3-yl)methoxy)benzonitrile. To a solution of 4-((3-methylazetidin-3-yl)methoxy)benzonitrile (60 mg, 0.30 mmol) and Et3N (85 μL, 0.61 mmol) in anhydrous DCM (3 mL) was added solid 2,4-dichlorobenzenesulfonyl chloride (80 mg, 0.33 mmol) and the mixture was stirred at rt under N2 for 18 h. The reaction mixture was concentrated in vacuo and the residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (106 mg) as a foam.1 H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.58-7.62(m,2H),7.57(d,1H),7.38(dd,1H),6.88-6.94(m,2H),4.02(d,2H),3.95(s,2H),3.79(d,2H),1.41(s,3H). LCMS[M+H] + 410.9,412.9.

[0227] Example 2 - Compound 2

[0228]

[0229] 3-((4-cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid was prepared according to Reaction Scheme 2:

[0230]

[0231] Reagents: Step 1) LHMDS, paraformaldehyde, THF, –78°C to rt; 2) DIAD, PPh3, 4-hydroxybenzonitrile, THF; 3) TFA, DCM, 2,4-dichlorobenzenesulfonyl chloride, NaOH, H2O, DCM.

[0232] Step 1.3-(hydroxymethyl)azetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester. A dry 3-neck 250mL flask equipped with an addition funnel is placed under N2 atmosphere and loaded with anhydrous THF (60mL). It is cooled to-78 ° C and LHMDS (1.0M, in THF, 11.8mL, 11.8mmol) is added. A solution of azetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-methyl ester (2.10g, 9.75mmol) in anhydrous THF (25mL) is added dropwise, and the mixture is stirred at-78 ° C and N2 for 1.5h. In a separate flask, paraformaldehyde (3.56g, 118.7mmol) is heated with a heating gun under N2, and the generated formaldehyde gas is bubbled into the reaction mixture. The reaction is warmed to rt while stirring for 18h. The reaction mixture was quenched with half-saturated NH4Cl (200 mL) and extracted with EtOAc (2 × 100 mL). The organic phase was washed with brine (200 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (268 mg) as a light yellow oil. 1H NMR (500MHz, CDCl3) δppm 4.15(d,2H), 4.04(d,2H), 3.91-3.98(m,2H), 3.80(s,3H), 2.27-2.36(m,1H), 1.44(s,9H).

[0233] Step 2. 1-(tert-butyl)3-methyl 3-((4-cyanophenoxy)methyl)azetidine-1,3-dicarboxylate. To a solution of 1-(tert-butyl)3-methyl 3-(hydroxymethyl)azetidine-1,3-dicarboxylate (268 mg, 1.09 mmol), 4-hydroxybenzonitrile (144 mg, 1.21 mmol) and PPh3 (430 mg, 1.64 mmol) in anhydrous THF (10 mL) was added neat DIAD (0.32 mL, 1.63 mmol) dropwise, and the mixture was stirred at rt under N2 for 27 h. The reaction mixture was diluted with EtOAc (50 mL), washed with saturated aqueous NaHCO3 (2×50 mL) and brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was transferred into EtOAc (50 mL), washed with KCO aqueous solution (3 x 50 mL) and brine (50 mL), dried over NaSO, filtered and concentrated in vacuo. The reaction mixture was purified by flash chromatography (hexane / EtOAc) to give the title compound (444 mg, -50% purity) as a colorless oil mixed with the DIAD by-product. 1 H NMR (500MHz, CDCl3) δppm 7.60 (d, 2H), 6.97 (d, 2H), 4.35 (s, 2H), 4.27 (d, 2H), 3.95 (d, 2H), 3.80 (s, 3H), 1.40-1.48 (m, 9H).

[0234] Step 3. 3-((4-cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid. A mixture of 1-(tert-butyl)3-methyl 3-((4-cyanophenoxy)methyl)azetidine-1,3-dicarboxylate (444 mg, 50% purity, 0.64 mmol), TFA (2 mL) and DCM (10 mL) was stirred at rt under N2 for 3 h. The reaction mixture was concentrated in vacuo and the residue was transferred to 1 M HCl (25 mL) and extracted with DCM (2×25 mL). The aqueous layer was made basic with 1 M NaOH (35 mL) and extracted with DCM (3×25 mL). The aqueous layer was concentrated in vacuo to a total volume of about 20 mL (with a lot of inorganic salt precipitation). Solid NaOH (49 mg, 1.23 mmol), DCM (10 mL) and water were added so as to stir well. Solid 2,4-dichlorobenzenesulfonyl chloride (324 mg, 1.32 mmol) was added once, and the biphasic mixture was stirred vigorously at rt and N2 for 7 days. 1M aqueous HCl solution (10 mL) was added, the reaction mixture was diluted with H2O until all salts were dissolved, and extracted with DCM (3×25 mL). The extract was dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (DCM / CH3OH / AcOH). The product fractions were concentrated in vacuo, toluene / CH3OH and Et2O were added and removed successively under vacuum to give the title compound (82 mg) as a white solid. 1 H NMR (500MHz, DMSO-d6) δppm 7.99(d,1H),7.96(d,1H),7.77(d,2H),7.68(dd,1H),7.06(d,2H),4.37(s,2H),4.23(d,2H),4.06(d,2H). LCMS[M+H] + 441.0,443.0.

[0235] Example 3 - Compound 3

[0236]

[0237] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)benzonitrile was prepared according to Reaction Scheme 3:

[0238]

[0239] Reagents: Step 1) BH3·THF, THF.

[0240] Step 1. BH3·THF solution (1.0 M in THF, 0.22 mL, 0.22 mmol) was added dropwise to 3-((4-cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (32 mg, 0.07 mmol) in anhydrous THF (2 mL) at 0°C and N2. The mixture was warmed to rt and stirred for 19 h. The reaction mixture was quenched with half-saturated NH4Cl (5 mL) and extracted with EtOAc (2×10 mL). The organic phase was washed with brine (10 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (8.9 mg) as a viscous colorless oil. 1 H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.60(d,2H),7.56(d,1H),7.39(dd,1H),6.93(d,2H),4.16(s,2H),3.96-4.01(m,2H),3.92-3.96(m,2H),3.91(s,2H). LCMS[M+H] + 427.0,429.0.

[0241] Example 4-Compound 4

[0242]

[0243] 3-((4-cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)-N-methylazetidine-3-carboxamide was prepared according to Reaction Scheme 4:

[0244]

[0245] Reagents: Step 1) HATU, CH3NH2, DIPEA, DMF.

[0246] Step 1. To a mixture of 3-((4-cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (14.7 mg, 0.033 mmol) and HATU (18.0 mg, 0.047 mmol) in anhydrous DMF (0.5 mL) was added a mixture of DIPEA (18 μL, 0.10 mmol) and methylamine (2.0 M in THF, 0.08 mL, 0.160 mmol) and the reaction mixture was stirred at rt for 4 h. The reaction mixture was diluted with EtOAc (2 mL), washed with saturated aqueous NaHCO3 (2×2 mL) and brine (2 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (7.2 mg) as a white solid. 1 H NMR (500MHz, CDCl3) δppm 7.98(d,1H),7.62(d,2H),7.58(d,1H),7.41(dd,1H),6.95(d,2H),6.14(br s,1H),4.33(s,2H),4.28(d,2H),4.09(d,2H),2.88(d,3H). LCMS[M+H] + 453.9,456.0.

[0247] Example 5-Compound 5

[0248]

[0249] 3-((4-Cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)-N-methyl-N-(2-(methylamino)ethyl)azetidine-3-carboxamide was prepared in a manner similar to Scheme 4. 1 H NMR (mixture of rotamers, 500 MHz, CDCl3) δ ppm 7.97 (d, 1H), 7.61 (d, 2H), 7.56 (d, 1H), 7.38 (dd, 1H), 6.96 (d, 2H), 4.53 and 4.47 (d, 2H), 4.44 and 4.37 (s, 2H), 3.96-4.06 (m, 2H), 3.43-3.52 (m, 2H), 3.14-3.22 (m, 1H), 2.96 and 2.92 (s, 3H), 2.69-2.81 (m, 2H), 2.39 (s, 3H). LCMS [M+H] + 511.0,513.0.

[0250] Example 6-Compound 6

[0251]

[0252] 3-((4-Cyanophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)-N-(tetrahydro-2H-pyran-4-yl)azetidine-3-carboxamide was prepared in a manner similar to Scheme 4. 1 HNMR(500MHz,CDCl3)δppm 7.98(d,1H),7.60-7.66(m,2H),7.58(d,1H),7.41(dd,1H),6.92-6.99(m,2H),5.99(d,1H),4.33(s,2H),4.27(d, 2H),4.10(d,2H),3.98-4.06(m,1H),3.90-3.97(m,2H),3.42-3.52(m,2H),1.84-1.93(m,2H),1.40-1.52(m,2H). LCMS[M+H] + 523.9,526.0.

[0253] Example 7-Compound 7

[0254]

[0255] Ethyl 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylate was prepared according to Reaction Scheme 5:

[0256]

[0257] Reagents: Step 1) Pd(OH)2 / C, HCl, EtOH; 2) 2,4-dichlorobenzenesulfonyl chloride, Et3N, DCM; 3) LiAlH(Ot-Bu)3, THF; 4) methanesulfonyl chloride, Et3N, DCM; 5) 2-fluoro-4-hydroxybenzonitrile, K2CO3, CH3CN, Δ.

[0258] Step 1. Azetidine-3,3-dicarboxylic acid diethyl ester hydrochloride. Pd(OH)2 (20 wt% on carbon, about 50% water, 1.548 g), 1-benzylazetidine-3,3-dicarboxylic acid diethyl ester (prepared according to Syn. Commun. 2003, 33, 3347–3353, 7.613 g, 26.12 mmol) and HCl (4.0 M in dioxane, 7.20 mL, 28.80 mmol) were stirred in EtOH (200 mL) under H2 atmosphere for 18 h. The mixture was filtered through a pad of celite, rinsed with CH3OH, and the filtrate was concentrated in vacuo to give the title compound (6.209 g) as a light yellow oil. 1H NMR (500MHz, CDCl3) δppm 10.06 (brs, 1H), 4.49 (br s, 4H), 4.31 (q, 4H), 1.26-1.35 (m, 6H).

[0259] Step 2. Diethyl 1-((2,4-dichlorophenyl)sulfonyl)azetidine-3,3-dicarboxylate. To a solution of diethyl azetidine-3,3-dicarboxylate hydrochloride (6.209 g, 26.12 mmol) in anhydrous DCM (250 mL) was added neat Et3N (11.5 mL, 82.50 mmol) and 2,4-dichlorobenzenesulfonyl chloride (8.15 g, 33.2 mmol), and the mixture was stirred at rt under N2 for 20 h. The reaction mixture was poured into saturated aqueous NaHCO3 (500 mL) and extracted with DCM (2×500 mL). The extract was dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (hexane / EtOAc) to give the title compound (8.80 g) as a colorless oil. 1 H NMR (500MHz, CDCl3) δppm 7.95 (d, 1H), 7.55 (d, 1H), 7.37 (dd, 1H), 4.40 (s, 4H), 4.25 (q, 4H), 1.27 (t, 6H).

[0260] Step 3. 1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidine-3-carboxylic acid ethyl ester. LiAlH(Ot-Bu)3 solution (1.0 M in THF, 45.5 mL, 45.5 mmol) was added dropwise to a solution of 1-((2,4-dichlorophenyl)sulfonyl)azetidine-3,3-dicarboxylic acid diethyl ester (8.8 g, 21.45 mmol) in anhydrous THF (200 mL) via an addition funnel at 0°C and N2. The mixture was warmed to rt and stirred for 22 h. The reaction mixture was diluted with EtOAc (500 mL), washed with 1M HCl (2×500 mL) and brine (500 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexanes / EtOAc) to give the title compound as a white crystalline solid (5.88 g). 1 H NMR(500MHz,CDCl3)δppm 7.93-8.00(m,1H),7.53-7.58(m,1H),7.35-7.40(m,1H),4.32(d,2H),4.20-4.27(m,2H),3.96(s,2H),3.92(d,2H),1.26-1.32(m,3H).

[0261] Step 4. Ethyl 1-((2,4-dichlorophenyl)sulfonyl)-3-(((methylsulfonyl)oxy)methyl)azetidine-3-carboxylate. To a solution of ethyl 1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidine-3-carboxylate (5.88 g, 15.97 mmol) and Et3N (3.4 mL, 24.4 mmol) in anhydrous DCM (200 mL) was added neat methanesulfonyl chloride (1.40 mL, 18.08 mmol) dropwise and the mixture was stirred at rt under N2 for 3 h. The reaction mixture was poured into saturated aqueous NaHCO3 (500 mL) and extracted with DCM (2×500 mL). The extract was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound (6.67 g) as an orange oil. 1 H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.56(d,1H),7.36-7.41(m,1H),4.55(s,2H),4.38(d,2H),4.25(q,2H),4.00(d,2H),3.07(s,3H),1.27-1.32(m,3H).

[0262] Step 5. Ethyl 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylate. A mixture of ethyl 1-((2,4-dichlorophenyl)sulfonyl)-3-(((methylsulfonyl)oxy)methyl)azetidine-3-carboxylate (5.33 g, 11.94 mmol), 2-fluoro-4-hydroxybenzonitrile (1.96 g, 14.33 mmol) and K2CO3 (3.30 g, 23.87 mmol) in anhydrous CH3CN (50 mL) was stirred at 80°C under N2 for 20 h. The reaction mixture was cooled, diluted with EtOAc (500 mL), washed with 2M K2CO3 (2×400 mL) and brine (2×400 mL), dried over Na2SO4, filtered and concentrated in vacuo to give the title compound (5.59 g, 11.47 mmol) as a viscous colorless oil. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.56-7.58(m,1H),7.52-7.55(m,1H),7.39(dd,1H),6.78(dd,1H), 6.73(dd,1H),4.41(d,2H),4.36(s,2H),4.25(q,2H),4.10(d,2H),1.27(t,3H). LCMS[M+H] + 487,488.

[0263] Example 8-Compound 8

[0264]

[0265] 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid was prepared according to Reaction Scheme 6:

[0266]

[0267] Reagents: Step 1) 4M NaOH aqueous solution, THF.

[0268] Step 1. To a solution of ethyl 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylate (1.02 g, 2.09 mmol) in THF (20 mL) was added 4M aqueous NaOH (5.30 mL, 21.2 mmol) and the mixture was stirred at rt under N2 for 3 h. The reaction mixture was concentrated in vacuo to remove THF, made acidic with 1M HCl (50 mL), diluted with water (50 mL), saturated with solid NaCl and extracted with DCM (3×100 mL). The extract was dried over Na2SO4, filtered and concentrated in vacuo to give the title compound (910 mg) as a white foam. 1 H NMR (500MHz, CDCl3) δppm 7.97(d,1H),7.53-7.59(m,2H),7.39(dd,1H),6.79(dd,1H),6.75(dd,1H),4.47(d,2H),4.39(s,2H),4.13(d,2H). LCMS[M+H] + 459,461.

[0269] Example 9-Compound 9

[0270]

[0271] 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxamide was prepared according to Reaction Scheme 7:

[0272]

[0273] Reagents: Step 1) HATU, NH3, DIPEA, DMF

[0274] Step 1. To a solution of 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (27.0 mg, 0.059 mmol) and HATU (28.5 mg, 0.075 mmol) in anhydrous DMF (2 mL) was added neat DIPEA (32 μL, 0.184 mmol) under N2, and the mixture was stirred at rt for 30 min. Ammonia gas was bubbled through the reaction mixture for 5 min, and the mixture was stirred at rt for 19 h. The reaction mixture was diluted with EtOAc (10 mL), washed with saturated NaHCO3 aqueous solution (2×10 mL) and brine (2×10 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (5.0 mg) as a yellow oil. 1 HNMR(500MHz, CDCl3)δppm 7.99(d,1H),7.55-7.61(m,2H),7.42(dd,1H),6.79(dd,1H),6.75(dd,1H),6.18(br s,br s,1H),5.60(br s,br s,1H),4.34(s,2H),4.28(d,2H),4.16(d,2H). LCMS[M+H] + 458.0,460.0.

[0275] Example 10-Compound 10

[0276]

[0277] (3-((4-(aminomethyl)-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methanol was prepared according to Reaction Scheme 8:

[0278]

[0279] Reagents: Step 1) BH3·THF, THF.

[0280] Step 1. To a solution of 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (248 mg, 0.54 mmol) in anhydrous THF (5 mL) was added dropwise BH3·THF solution (1.0 M in THF, 1.65 mL) at 0°C and N2. The mixture was warmed to rt and stirred for 21 h. The reaction mixture was quenched with 1M HCl (10 mL), neutralized with saturated NaHCO3 aqueous solution (35 mL), and extracted with EtOAc (2×35 mL). The organic phase was washed with brine (35 mL), dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (EtOAc / hexanes, then DCM / CH3OH / NH4OH) to give the title compound (111 mg) as a white foam. 1 H NMR(500MHz,DMSO-d6)δppm 7.96(d,1H),7.95(d,1H),7.67(dd,1H),7.32(t,1H),6.55-6.61(m,2H),5.0 6(t,1H),3.93(s,2H),3.76-3.82(m,4H),3.64(s,2H),3.52(d,2H),1.98(br s,2H). LCMS[M+H] + 449.

[0281] Example 11-Compound 11

[0282]

[0283] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared according to Reaction Scheme 9:

[0284]

[0285] Reagents: Step 1) BH3.SMe2 (1.0 M in THF), THF.

[0286] Step 1. BH3·SMe2 solution (0.30 mL, 3.16 mmol) was added dropwise to 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (971 mg, 2.12 mmol) in anhydrous THF (20 mL) at 0°C and N2. The mixture was warmed to rt and stirred for 20 h. The reaction mixture was quenched with 1 M NaOH (15 mL), poured into saturated NaHCO3 aqueous solution (100 mL), and extracted with EtOAc (2×75 mL). The organic phase was washed with brine (75 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (760 mg, 1.71 mmol) as a viscous colorless oil. 1 H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.57(d,1H),7.53(dd,1H),7.39(dd,1H),6.75(dd,1H),6.71(dd,1 H),4.16(s,2H),3.95-3.99(m,2H),3.92-3.95(m,2H),3.90(d,2H),1.75(t,1H). LCMS[M+H] + 445,447. LC / MS / MS [MH] - 442.7.

[0287] Example 12-Compound 12

[0288]

[0289] (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl acetate was prepared according to Reaction Scheme 10:

[0290]

[0291] Reagents: Step 1) Acetic anhydride, Et3N, DMAP, DCM.

[0292] Step 1. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (19.7 mg, 0.044 mmol), Et3N (12 μL, 0.088 mmol) and acetic anhydride (10 μL, 0.088 mmol) in anhydrous DCM (0.5 mL) was added catalytic DMAP and the mixture was stirred at rt for 6 h. The reaction mixture was directly purified by flash chromatography on silica gel (EtOAc / hexanes) to give the title compound (20.1 mg) as a viscous colorless oil. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.52-7.59(m,2H),7.39(dd,1H),6.76(dd,1H),6.71(dd,1H),4.32(s,2H),4.12(s,2H),4.01(d,2H),3.96(d,2H),2.07(s,3H). LCMS[M+H] + 487,489.

[0293] Example 13-Compound 13

[0294]

[0295] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(methoxymethyl)azetidin-3-yl)methoxy)-fluorobenzonitrile was prepared according to Reaction Scheme 11:

[0296]

[0297] Reagents: Step 1) NaH, CH3I, THF.

[0298] Step 1. NaH (9.2 mg, 0.383 mmol) was added to an oven-dried flask equipped with a stirring bar, the flask was capped and placed under N2 atmosphere, and anhydrous THF (0.5 mL) was added. A solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (53.2 mg, 0.119 mmol) in anhydrous THF (0.5 mL) was added dropwise, followed by a solution of CH3I (16.0 μL, 0.257 mmol) in anhydrous THF (0.5 mL), and the mixture was stirred at rt for 20 h. The reaction mixture was diluted with EtOAc (15 mL) and washed with saturated NaHCO3 aqueous solution (2×10 mL) and brine (2×10 mL). The organic layer was dried over Na2SO4, decanted and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (hexanes / EtOAc) to give the title compound as a white solid (17.2 mg). 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.56-7.59(m,1H),7.53(dd,1H),7.39(dd,1H),6.74(dd,1H),6.69 (dd,1H),4.09(s,2H),3.97(d,2H),3.88-3.92(m,2H),3.57(s,2H),3.34(s,3H).

[0299] Example 14-Compound 14

[0300]

[0301] Sodium (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl phosphate was prepared according to Reaction Scheme 12:

[0302]

[0303] Reagents: Step 1) Phosphorus oxychloride (V), Et3N, THF.

[0304] Step 1. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (89 mg, 0.20 mmol) and Et3N (56 μL, 0.40 mmol) in anhydrous THF (2.0 mL) was added neat phosphorus oxychloride (V) (30 μL, 0.32 mmol) dropwise, and the mixture was stirred at rt for 3 h. The reaction mixture was quenched with 1 M NaOH (5 mL), diluted with H2O (10 mL) and washed with EtOAc (2×15 mL). The aqueous layer was acidified to pH 1 with 6 M HCl and extracted with DCM (3×20 mL). The extract was dried over Na2SO4, filtered and concentrated to dryness to give the phosphate (83 mg, 0.16 mmol). The residue was dissolved in CH3OH and THF, and 0.1M aqueous NaHCO3 (3.2 mL) was added. The mixture was concentrated to dryness, and the residue was concentrated twice from anhydrous CH3OH to give the title compound (85 mg, 0.15 mmol) as a white solid. 1 H NMR(500MHz,CD3OD)δppm 8.03(d,1H),7.75(d,1H),7.63(dd,1H),7.56(dd,1H),6.92(dd,1H),6.85(dd,1H),4.17(s,2H),4.01-4.08(m,4H),3.91(d,2H). LCMS[M+H] + 524.9,526.9. LC / MS / MS [MH] - 522.8.

[0305] Example 15-Compound 15

[0306]

[0307] 2-Aminoethane-1-sulfonic acid (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl ester trifluoroacetate was prepared according to Reaction Scheme 13:

[0308]

[0309] Reagents: Step 1) N-Boc taurine chloride, Et3N, DCM; 2) TFA, DCM.

[0310] Step 1. (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl 2-((tert-butoxycarbonyl)amino)ethane-1-sulfonate. To a mixture of N-Boc taurine hydrochloride (24 mg, 0.10 mmol) prepared as described in Nucleosides, Nucleotides, and Nucleic Acids 2013, 32, 599 in anhydrous DCM (0.5 mL) was added a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (34 mg, 0.077 mmol) and Et3N (22 μL, 0.16 mmol) in anhydrous DCM (1 mL) and the mixture was stirred at rt for 3 days. The reaction mixture was purified directly by flash chromatography (EtOAc / hexanes) to give the title compound as a colorless oil (29 mg).

[0311] Step 2. 2-Aminoethane-1-sulfonic acid (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl ester trifluoroacetate. To a solution of 2-((tert-butoxycarbonyl)amino)ethane-1-sulfonic acid (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl ester (29 mg, 0.044 mmol) in anhydrous DCM (1 mL) was added neat TFA (0.1 mL) and the mixture was stirred at rt for 2 h. The reaction mixture was concentrated to dryness and purified by HPLC from dihydrogen phosphate. The mixture was concentrated to dryness twice with oxane and Et2O to give the title compound as a viscous yellow oil (30 mg). 1 H NMR(500MHz,DMSO-d6)δppm 7.91-8.06(m,5H),7.85(t,1H),7.67(dd,1H),7.06(dd,1H),6.88(dd,1H),4 .50(s,2H),4.21(s,2H),3.89-3.98(m,4H),3.69(t,2H),3.15-3.24(m,2H). LCMS[M+H] + 552.0,554.1.

[0312] Example 16-Compound 16

[0313]

[0314] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((methylamino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared according to Reaction Scheme 14:

[0315]

[0316] Reagents: Step 1) Et3N, oxalyl chloride, DMSO, DCM; –78°C; 2) CH3NH2, NaBH(OAc)3, AcOH, DCE.

[0317] Step 1. 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile. To a solution of oxalyl chloride (0.350 mL, 4.027 mmol) in anhydrous DCM (3 mL) was added dropwise a solution of DMSO (0.560 mL, 7.887 mmol) in anhydrous DCM (3 mL) at -78 °C under N2. The mixture was stirred for 10 min and a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (760 mg, 1.707 mmol) in anhydrous DCM (10 mL) was added dropwise. The mixture was stirred at -78 °C for 30 min and Et3N (1.30 mL, 9.326 mmol) was added dropwise. The mixture was warmed to rt and stirred for 22 h. The reaction mixture was poured into saturated NaHCO aqueous solution (100 mL) and extracted with DCM (3×50 mL). The extract was dried over NaSO, filtered and concentrated in vacuo to give the crude title compound (681.1 mg) as a yellow oil. 1 H NMR(500MHz,CDCl3)δppm 9.87-9.93(m,1H),7.96-8.00(m,1H),7.51-7.60(m,2H),7.39-7.43(m,1H),6.7 6-6.80(m,1H),6.72-6.76(m,1H),4.38(s,2H),4.30(s,2H),4.15-4.20(m,2H).

[0318] Step 2. 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((methylamino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile (258 mg, 0.58 mmol), methylamine (2.0 M in THF, 0.73 mL, 1.46 mmol) and acetic acid (2 drops) in anhydrous DCE (5 mL) was added solid NaBH(OAc)3 (310 mg, 1.46 mmol) and the mixture was stirred at rt for 22 h. The reaction mixture was poured into saturated aqueous NaHCO3 (100 mL) and extracted with DCM (3×50 mL). The extract was dried over Na2SO4, decanted and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (hexanes / EtOAc / CH3OH). To the purified material was added HCl solution (4.0 M in distilled water). The solid was triturated with CH3OH / Et2O and the precipitate was collected by vacuum filtration to give the title compound as a white solid (106 mg). 1 HNMR(500MHz,DMSO-d6)δppm 8.79(br s,br s,2H),7.97(m,2H),7.86(m,1H),7.67(dd,1H),7.01(dd,1H),6.87(dd,1H),4.23(s,2H),4.02(d,2H),3.87(d,2H),3.33(m,2H),2.58(brs,br s,3H). LCMS[M+H] + 458,460.

[0319] Example 17-Compound 17

[0320]

[0321] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-hydroxyethyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and ethanolamine. 1H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.57(d,1H),7.54(dd,1H),7.39(dd,1H),6.75(dd,1H),6.72(dd,1H), 4.13(s,2H),3.89-3.97(m,4H),3.62-3.67(m,2H),2.97(s,2H),2.77-2.83(m,2H). LCMS[M+H] + 488,490.

[0322] Example 18-Compound 18

[0323]

[0324] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((tetrahydro-2H-pyran-4-yl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 4-aminotetrahydropyran. 1 H NMR (free base) (500MHz, CDCl3) δppm 7.98(d,1H),7.57(d,1H),7.53(dd,1H),7.39(dd,1H),6.74(dd,1H),6.71(dd,1H),4.11-4.13(m,2H),3.90- 3.97(m,4H),3.85-3.90(m,2H),3.31-3.40(m,2H),2.95(s,2H),2.55-2.66(m,1H),1.78(d,2H),1.30(d,2H). LCMS[M+H] + 528,530.

[0325] Example 19-Compound 19

[0326]

[0327] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((3-hydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 3-amino-1-propanol. 1H NMR (free base) (500 MHz, CDCl3) δ ppm 7.97 (d, 1H), 7.56 (d, 1H), 7.54 (dd, 1H), 7.37-7.40 (m, 1H), 6.75 (dd, 1H), 6.71 (dd, 1H), 4.09-4.12 (m, 2H), 3.87-3.96 (m, 4H), 3.73-3.78 (m, 2H), 2.97 (s, 2H), 2.86 (t, 2H), 1.70 (quin, 2H). LCMS [M+H] + 502,504.

[0328] Example 20-Compound 20

[0329]

[0330] 2-(((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)amino)acetamide hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and glycinamide hydrochloride. 1 H NMR (free base) (500 MHz, CDCl3) δ ppm 7.97 (d, 1H), 7.52-7.58 (m, 2H), 7.39 (dd, 1H), 6.76 (dd, 1H), 6.72 (dd, 1H), 6.48 (br s, 1H), 5.41-5.49 (m, 1H), 4.13-4.16 (m, 2H), 3.97-4.02 (m, 2H), 3.90-3.95 (m, 2H), 3.34 (s, 2H), 2.97 (s, 2H). LCMS [M+H] + 501,503.

[0331] Example 21-Compound 21

[0332]

[0333] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-methoxyethyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 2-methoxyethylamine. 1HNMR (free base) (500MHz, CDCl3) δppm 7.97 (d, 1H), 7.56 (d, 1H), 7.50-7.55 (m, 1H), 7.38 (dd, 1H), 6.68-6.77 (m, 2H), 4.13 (s, 2H), 3.91-3.96 (m, 2H), 3.87 (d, 2H), 3.43 (t, 2H), 3.32 (s, 3H), 2.94 (s, 2H), 2.76 (t, 2H). LCMS [M+H] + 502,504.

[0334] Example 22-Compound 22

[0335]

[0336] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((thiazol-2-ylmethyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and thiazol-2-ylmethanamine. 1 HNMR (free base) (500MHz, CDCl3) δppm 8.10(s,1H),7.97(d,1H),7.70(d,1H),7.55(d,1H),7.54–7.51(m,1H),7.38(dd,1H),6.7 4(dd,1H),6.70(dd,1H),4.17(s,2H),4.15(s,2H),3.96(d,2H),3.92(d,2H),3.04(s,2H). LCMS[M+H] + 541,543.

[0337] Example 23-Compound 23

[0338]

[0339] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((tetrahydrofuran-3-yl)methyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and (tetrahydrofuran-3-yl)methanamine. 1H NMR (free base) (500MHz, CDCl3) δppm 7.91(d,1H),7.44-7.52(m,2H),7.32(dd,1H),6.61-6.71(m,2H),4.05(s,2H),3.80-3.89(m,4H),3.70-3.78(m,2H), 3.62-3.68(m,1H),3.41(dd,1H),2.86(q,2H),2.47-2.60(m,2H),2.18-2.28(m,1H),1.87-1.99(m,1H),1.47(dd,1H). LCMS[M+H] + 528,530.

[0340] Example 24-Compound 24

[0341]

[0342] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-hydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 1-amino-2-propanol. 1 H NMR (free base) (500 MHz, CDCl3) δ ppm 7.91 (d, 1H), 7.43-7.53 (m, 2H), 7.32 (dd, 1H), 6.62-6.72 (m, 2H), 4.07 (s, 2H), 3.83-3.90 (m, 4H), 3.67-3.75 (m, 1H), 2.85-2.97 (m, 2H), 2.66 (dd, 1H), 2.39 (dd, 1H), 1.08 (d, 3H). LCMS [M+H] + 502,504.

[0343] Example 25-Compound 25

[0344]

[0345] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((ethylamino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a manner similar to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and ethylamine (2.0 M in THF). 1H NMR (free base) (500 MHz, CDCl3) δ ppm 7.97 (d, 1H), 7.50-7.59 (m, 2H), 7.39 (dd, 1H), 6.69-6.78 (m, 2H), 4.12 (s, 2H), 3.92-3.96 (m, 2H), 3.86-3.90 (m, 2H), 2.93 (s, 2H), 2.64 (q, 2H), 1.06 (t, 3H). LCMS [M+H] + 472,474.

[0346] Example 26-Compound 26

[0347]

[0348] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((oxetan-3-ylamino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and oxetan-3-amine. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.52-7.58(m,2H),7.39(dd,1H),6.75(dd,1H),6.71(dd,1H),4 .80(t,2H),4.37(t,2H),4.12-4.15(m,2H),3.85-3.96(m,5H),2.90(s,2H). LCMS[M+H] + 500,502.

[0349] Example 27-Compound 27

[0350]

[0351] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((oxetan-3-ylmethyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 3-oxetanylmethylamine. 1H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.51-7.58(m,2H),7.36-7.40(m,1H),6.74(dd,1H),6.70(dd,1H),4.76(dd ,2H),4.34(t,2H),4.11(s,2H),3.85-3.94(m,4H),2.97-3.08(m,1H),2.91-2.97(m,4H). LCMS[M+H] + 514,516.

[0352] Example 28-Compound 28

[0353]

[0354] (S)-4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((tetrahydrofuran-2-yl)methyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and (S)-tetrahydrofuranmethylamine. 1 H NMR(500MHz,DMSO-d6)δppm 9.21(br s,1H),9.04(br s,1H),7.97(dd,1H),7.86(t,1H),7.67(dd,2H),6.98(dd,1H),6.84(dd,1H),4.23-4.29(m,2H ),3.98(d,2H),3.90(d,2H),3.74-3.82(m,1H),3.63-3.71(m,1H),3.30-3.48(m,3H),3.10(br s,1H),2.94(br s,1H),1.93-2.04(m,1H),1.75-1.90(m,2H),1.49-1.59(m,1H). LCMS [M+H] + 528,530.

[0355] Example 29-Compound 29

[0356]

[0357] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((tetrahydro-2H-pyran-4-yl)methyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 4-aminomethyltetrahydropyran. 1 H NMR(500MHz,DMSO-d6)δppm 9.01-8.82(m,2H),8.03-7.95(m,2H),7.91-7.83(m,1H),7.69(dd,1H),6.99(dd,1H),6.85(dd,1H),4.29(s,2H),4.03(d,2H),3.92(d ,2H),3.89-3.81(m,2H),3.43-3.37(m,2H),3.26(td,2H),2.93-2.84(m,2H),2.11-1.98(m,1H),1.73-1.63(m,2H),1.30-1.14(m,2H). LCMS[M+H] + 542,544.

[0358] Example 30-Compound 30

[0359]

[0360] (R)-4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-hydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a manner similar to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and (R)-(-)-1-amino-2-propanol. 1 H NMR(500MHz,DMSO-d6)δppm 8.95-8.78(m,2H),8.02-7.96(m,2H),7.90-7.85(m,1H),7.69(dd,1H),7.01(dd,1H),6.87(dd,1H),5.50-5.29(m,1H),4 .28(s,2H),4.09-3.97(m,3H),3.94-3.88(m,2H),3.45-3.37(m,2H),3.07-2.98(m,1H),2.86-2.76(m,1H),1.11(d,3H). LCMS[M+H] + 502,504.

[0361] Example 31-Compound 31

[0362]

[0363] (S)-4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-hydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a manner similar to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and (S)-(+)-1-amino-2-propanol. 1 H NMR(500MHz,DMSO-d6)δppm 8.96-8.78(m,2H),8.01-7.96(m,2H),7.91-7.85(m,1H),7.69(dd,1H),7.01(dd,1H),6.87(dd,1H),5.50-5.30(m,1H),4 .28(s,2H),4.08-3.97(m,3H),3.94-3.88(m,2H),3.46-3.39(m,2H),3.07-2.99(m,1H),2.86-2.76(m,1H),1.11(d,3H). LCMS[M+H] + 502,504.

[0364] Example 32-Compound 32

[0365]

[0366] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((1-hydroxypropan-2-yl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and (+ / -) alaninol. 1H NMR(500MHz,DMSO-d6)δppm8.90-8.65(m,2H),8.03-7.96(m,2H),7.91-7.85(m,1H),7.69(dd,1H),6.99(dd,1H),6.86(dd,1H),5.41(s,1H), 4.27(s,2H),4.02-3.96(m,2H),3.96-3.90(m,2H),3.73-3.64(m,2H), 3.59-3.52(m,1H),3.52-3.45(m,1H),3.37-3.28(m,1H),1.23(d,3H). LCMS[M+H] + 502,504.

[0367] Example 33-Compound 33

[0368]

[0369] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2,3-dihydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 3-amino-1,2-propanediol. 1 H NMR(500MHz,CD3OD)δppm8.05-8.00(m,1H),7.77(d,1H),7.71(dd,1H),7.58-7.54(m,1H),7.01(dd,1H),6.97(dd,1H),4.39-4.33(m,2H ),4.10-4.02(m,4H),4.01-3.95(m,1H),3.68-3.64(m,1H),3.64-3.62(m,1H),3.62-3.55(m,2H),3.28-3.26(m,1H),3.20-3.14(m,1H). LCMS[M+H] + 518,520.

[0370] Example 34-Compound 34

[0371]

[0372] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((3-hydroxy-2-methylpropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 3-amino-2-methylpropan-1-ol. 1 H NMR(500MHz,CD3OD)δppm 8.02(d,1H),7.76(d,1H),7.69(dd,1H),7.56(dd,1H),6.99(dd,1H),6.95(dd,1H),4.35(s,2H),3.98-4.11(m,4 H),3.71-3.78(m,1H),3.54-3.59(m,1H),3.43-3.51(m,2H),3.06-3.17(m,2H),2.16-2.27(m,1H),0.93(d,3H). LCMS[M+H] + 516,518.

[0373] Example 35-Compound 35

[0374]

[0375] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((3-hydroxybutyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 4-amino-2-butanol. 1 H NMR(500MHz,CD3OD)δppm 8.02(d,1H),7.77(d,1H),7.70(dd,1H),7.53-7.58(m,1H),6.98(dd,2H),6.94(dd,2H),4.32(s,2H),4.00-4 .08(m,4H),3.95(s,1H),3.44-3.57(m,2H),3.20-3.27(m,2H),1.84(d,1H),1.77(s,1H),1.18-1.24(m,3H). LCMS[M+H] + 516,518.

[0376] Example 36-Compound 36

[0377]

[0378] 4-((3-(((2-cyanoethyl)amino)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and 3-aminopropionitrile. 1 H NMR(500MHz,CDCl3)δppm 7.96-7.99(m,1H),7.56-7.58(m,1H),7.51-7.55(m,1H),7.37-7.41(m,1H),6.74-6.77(m,1H),6.7 0-6.74(m,1H),4.11(s,2H),3.85-3.96(m,4H),2.93(s,2H),2.84-2.88(m,2H),2.44-2.48(m,2H). LCMS[M+H] + 497,499.

[0379] Example 37-Compound 37

[0380]

[0381] Ethyl 3-(((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)amino)propanoate hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and ethyl β-alaninate hydrochloride and NaOAc instead of AcOH. 1 H NMR(500MHz,DMSO-d6)δppm 8.91-9.08(m,2H),7.94-8.04(m,2H),7.87(t,1H),7.67(d,1H),7.00(d,1H),6.86(d,1H),4.25(s,2H),4.08( q,2H),4.01(d,2H),3.88(d,2H),3.35-3.46(m,2H),3.13-3.28(m,2H),2.77-2.88(m,2H),1.15-1.26(m,3H). LCMS[M+H] + 544.0,546.1.

[0382] Example 38-Compound 38

[0383]

[0384] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((cis)-3-hydroxycyclobutyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and cis-3-aminocyclobutanol hydrochloride and NaOAc. 1 H NMR(500MHz,CD3OD)δppm 8.02(d,1H),7.77(d,1H),7.71(dd,1H),7.56(dd,1H),6.98(dd,1H),6.93(dd,1H),4.29(s,2H) ,4.07-4.14(m,1H),3.98-4.06(m,4H),3.33-3.43(m,3H),2.67-2.78(m,2H),2.01-2.14(m,2H). LCMS[M+H] + 514.1,516.2.

[0385] Example 39-Compound 39

[0386]

[0387] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((cis)-4-hydroxycyclohexyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and cis-4-aminocyclohexanol hydrochloride and NaOAc. 1 H NMR(500MHz,CD3OD)δppm 8.03(d,1H),7.78(d,1H),7.68-7.74(m,1H),7.57(dd,1H),6.97(dd,1H),6.93(dd,1H),4.29(s,2H),4.05-4.09(m ,2H),3.98-4.04(m,3H),3.54(s,2H),3.13-3.23(m,1H),1.88-1.96(m,4H),1.78-1.87(m,2H),1.54-1.65(m,2H). LCMS[M+H] + 542.1,544.1.

[0388] Example 40-Compound 40

[0389]

[0390] (R)-4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((tetrahydrofuran-2-yl)methyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a similar manner to that in Scheme 14 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and [(2R)-oxolan-2-yl]methanamine hydrochloride and NaOAc. 1 H NMR(500MHz,DMSO-d6+K2CO3)δppm 7.96-8.00(m,2H),7.80-7.86(m,1H),7.68(dd,1H),7.05(dd,1H),6.86(dd,1H),4.11(s,2H),3.77-3.8 4(m,4H),3.64-3.77(m,3H),3.52-3.61(m,2H),2.68-2.78(m,2H),1.68-1.88(m,3H),1.39-1.49(m,1H). LCMS[M+H] + 528.1,530.1.

[0391] Example 41-Compound 41

[0392]

[0393] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((trans)-4-hydroxycyclohexyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared according to Reaction Scheme 15:

[0394]

[0395] Reagents: Step 1) NaBH3CN, trans-4-aminocyclohexanol, CH3OH.

[0396] Step 1. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile (50.1 mg, 0.113 mmol) in CH3OH (1 mL) was added trans-4-aminocyclohexanol (18.3 mg, 0.159 mmol) and stirred until completely dissolved. NaBH3CN (12.8 mg, 0.204 mmol) was added and the mixture was stirred at rt for 18 h. The reaction mixture was poured into 2M K2CO3 (15 mL) and extracted with DCM (3×10 mL). The extract was dried over Na2SO4, decanted and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (hexane / EtOAC / CH3OH) to give the title compound (21.9 mg) as a white solid. 1 H NMR(500MHz,DMSO-d6)δppm8.81-8.71(m,2H),8.02-7.96(m,2H),7.88(dd,1H) ,7.69(dd,1H),7.00(dd,1H),6.86(dd,1H),4.26(s,2H),3.98(d,2H),3.91(d, 2H),3.73-3.69(m,1H),3.69-3.64(m,1H),3.52-3.48(m,1H),3.48-3.44(m,1H ),2.09-2.00(m,2H),1.92-1.84(m,2H),1.50-1.37(m,2H),1.21-1.10(m,2H). LCMS[M+H] + 542,544.

[0397] Example 42-Compound 42

[0398]

[0399] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-((((trans)-3-hydroxycyclobutyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared in a manner similar to that in Scheme 15 from 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-formylazetidin-3-yl)methoxy)-2-fluorobenzonitrile and trans-3-aminocyclobutan-1-ol. 1H NMR(500MHz,CD3OD)δppm 8.02(d,1H),7.77(d,1H),7.71(dd,1H),7.56(dd,1H),6.98(dd,1H),6.93(dd,1H),4.44-4.51(m ,1H),4.29(s,2H),4.01-4.08(m,4H),3.93-4.00(m,1H),3.39(s,2H),2.47-2.56(m,2H),2.35(br s,2H). LCMS[M+H] + 514,516.

[0400] Example 43-Compound 43

[0401]

[0402] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-hydroxyethyl)(methyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared according to Reaction Scheme 16:

[0403]

[0404] Reagents: Step 1) NaBH(OAc)3, 37 wt% formaldehyde, AcOH, DCE.

[0405] Step 1. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-hydroxyethyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (15 mg, 0.031 mmol), formaldehyde (37 wt % in H2O, 16 μL, 0.16 mmol) and acetic acid (1 drop) in anhydrous DCE (1 mL) was added solid NaBH(OAc)3 (16.9 mg, 0.0798 mmol) and the mixture was stirred at rt for 16 h. The reaction mixture was poured into saturated aqueous NaHCO3 (15 mL) and extracted with DCM (3×10 mL). The extract was dried over Na2SO4, decanted and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (hexane / EtOAc / CH3OH) to give the title compound (7.7 mg) as a viscous colorless oil. 1H NMR (free base) (500MHz, CDCl3) δppm 7.97(d,1H),7.56(d,1H),7.54(dd,1H),7.38(dd,1H),6.77(dd,1H),6.73(dd,1H),4.20(s ,2H),3.98(d,2H),3.90(d,2H),3.56(t,2H),2.79(s,2H),2.52-2.58(m,2H),2.22(s,3H). LCMS[M+H] + 502,504.

[0406] Example 44-Compound 44

[0407]

[0408] 4-((3-(Aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared according to Reaction Scheme 17:

[0409]

[0410] Reagents: Step 1) methanesulfonyl chloride, Et3N, DCM; 2) NaN3, DMF; 3) Zn, NH4Cl, CH3OH.

[0411] Step 1. (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (760 mg, 1.71 mmol) and Et3N (0.36 mL, 2.58 mmol) in anhydrous DCM (20 mL) was added neat methanesulfonyl chloride (0.16 mL, 2.06 mmol) dropwise, and the mixture was stirred at rt under N2 for 1 h. The reaction mixture was poured into saturated aqueous NaHCO3 (100 mL) and extracted with DCM (2×100 mL). The extract was dried over Na2SO4, filtered and concentrated to dryness to give the title compound (909 mg) as an off-white foam.

[0412] Step 2. 4-((3-(azidomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile. A mixture of (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate (909 mg, 1.71 mmol) and NaN3 (566 mg, 8.71 mmol) in anhydrous DMF (5 mL) was stirred at 50°C for 16 h. The reaction mixture was cooled, diluted with EtOAc (75 mL), washed with saturated aqueous NaHCO3 solution (2×50 mL) and brine (50 mL), dried over Na2SO4, filtered and concentrated to dryness to give the title compound (755 mg) as an almost colorless oil. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.52-7.60(m,2H),7.36-7.42(m,1H),6.68-6.80(m,2H), 4.08-4.12(m,2H),3.94-3.99(m,2H),3.88-3.93(m,2H),3.74(s,2H).

[0413] Step 3. 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-yl)methoxy)-2-fluorobenzonitrile. A mixture of 4-((3-(azidomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-yl)methoxy)-2-fluorobenzonitrile (755 mg, 1.61 mmol), NH4Cl (437 mg, 8.17 mmol) and zinc powder (2.09 g, 32.0 mmol) in CH3OH (20 mL) was stirred at rt under N2 for 3 h. The reaction mixture was diluted with DCM (about 20 mL) and filtered through celite, rinsing with DCM / CH3OH. The filtrate was concentrated to dryness and purified by flash chromatography (hexane / EtOAc / CH3OH) to give the title compound (583 mg) as a white solid. 1 H NMR(500MHz,DMSO-d6)δppm7.92-8.01(m,2H),7.84(t,1H),7.63-7.70(m,1H),6.99(dd,1H),6 .83(dd,1H),5.07(br.s,2H),4.12(s,2H),3.84-3.90(m,2H),3.78-3.83(m,2H),2.96(s,2H). LCMS[M+H] + 444,446.

[0414] Example 45-Compound 45

[0415]

[0416] N-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)acetamide was prepared as described below. To a solution of 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (8.9 mg, 0.020 mmol) and Et3N (15 μL, 0.108 mmol) in anhydrous DCM (0.5 mL) was added acetyl chloride (8 μL, 0.113 mmol) dropwise, and the mixture was stirred at rt for 2 h. The reaction mixture was poured into saturated NaHCO3 aqueous solution (5 mL) and extracted with DCM (2×5 mL). The extract was dried over Na2SO4, decanted and concentrated to dryness. The residue was purified by flash chromatography on silica gel (hexanes / EtOAc) to give the title compound as a colorless oil (3.8 mg). 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.57(d,1H),7.54(dd,1H),7.40(dd,1H),6.74(dd,1H),6.70(dd ,1H),5.85(t,1H),4.06(s,2H),3.91-3.99(m,4H),3.60(d,2H),2.02(s,3H). LCMS[M+H] + 486,488.

[0417] Example 46-Compound 46

[0418]

[0419] 2-Amino-N-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)acetamide was prepared according to Reaction Scheme 18:

[0420]

[0421] Reagents: Step 1) Boc-Glycine, HATU, DIPEA, DMF; 2) TFA, DCM.

[0422] Step 1. tert-Butyl (2-(((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)amino)-2-ketoethyl)carbamate. A mixture of 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (43 mg, 0.10 mmol), Boc-glycine (21 mg, 0.12 mmol), HATU (46 mg, 0.12 mmol) and DIPEA (60 μL, 0.34 mmol) in anhydrous DMF (1 mL) was stirred at rt for 18 h. The reaction mixture was diluted with EtOAc (10 mL), washed with saturated aqueous NaHCO3 (2×10 mL) and brine (2×10 mL), dried over Na2SO4, decanted and concentrated. The residue was purified by flash chromatography (EtOAc / hexanes) to give the title compound which was used immediately in the next step.

[0423] Step 2. 2-amino-N-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)acetamide. A mixture of tert-butyl (2-(((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)amino)-2-ketoethyl)carbamate (0.10 mmol) and TFA (0.25 mL) in anhydrous DCM (0.75 mL) was stirred at rt for 2 h and concentrated. The residue was added to saturated aqueous NaHCO3 solution (10 mL) and extracted with DCM (3×10 mL). The extracts were dried over Na2SO4, decanted and concentrated, and the residue was purified by flash chromatography (DCM / CH3OH / NH4OH) to give the title compound as a white solid (20 mg). 1 H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.87-7.94(m,1H),7.51-7.59(m,2H),7.39(dd,1H),6.76(dd,1H),6.71( dd,1H),4.08(s,2H),3.97-4.01(m,2H),3.92-3.97(m,2H),3.63(d,2H),3.39(s,2H). LCMS[M+H] + 501.1,503.1.

[0424] Example 47-Compound 47

[0425]

[0426] N-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azacyclo

[0427] Prepared from 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and glycolic acid. 1 H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.56(d,1H),7.52-7.55(m,1H),7.36-7.41(m,1H),6.96-7.03(m,1H),6.76(dd,1H ),6.72(dd,1H),4.18(d,2H),4.10(s,2H),4.03(d,2H),3.95(d,2H),3.67(d,2H),2.42(t,1H). LCMS[M+H] + 502.0,504.0.

[0428] Example 48-Compound 48

[0429]

[0430] N-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)-2-(methylamino)acetamide was prepared in a manner similar to that in Scheme 18 from 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and Boc-sarcosine. 1 HNMR(500MHz, CDCl3)δppm7.96(d,1H),7.73-7.81(m,1H),7.51-7.58(m,2H),7.38(dd,1H),6.75(dd ,1H),6.71(dd,1H),4.08(s,2H),3.92-3.99(m,4H),3.63(d,2H),3.27(s,2H),2.42(s,3H),1.75(br s,1H). LCMS[M+H] + 515.0,517.1.

[0431] Example 49-Compound 49

[0432]

[0433] 2-Amino-N-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)ethane-1-sulfonamide was prepared in a manner similar to that in Scheme 13 from 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and Boc-taurine chloride. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.57(d,1H),7.52-7.56(m,1H),7.40(dd,1H),6.75(dd,1H),6.70(dd,1H),5.76-5 .94(m,2H),4.16(s,2H),3.91-4.00(m,4H),3.45(s,2H),3.24-3.31(m,2H),3.07-3.13(m,2H). LCMS[M+H] + 550.9,553.0.

[0434] Example 50-Compound 50

[0435]

[0436] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(morpholinomethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared according to Reaction Scheme 19:

[0437]

[0438] Reagents: Step 1) 2-bromoethyl ether, K2CO3, CH3CN, 85°C.

[0439] Step 1. To 4-((3-(aminomethyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile (44 mg, 0.10 mmol) and K2CO3 was added a solution of 2-bromoethyl ether (19 μL, 0.15 mmol) in anhydrous CH3CN (1.0 mL), and the mixture was stirred in a capped bottle at 85 ° C for 16 h. The reaction mixture was cooled, diluted with EtOAc (10 mL), washed with saturated aqueous NaHCO3 (2×10 mL) and brine (10 mL), dried over Na2SO4, decanted and concentrated. The residue was purified by flash chromatography (EtOAc / hexanes). The product fractions were concentrated and purified by distillation. 4.0 M HCl in oxane was converted to the HCl salt and concentrated to dryness to give the title compound (33 mg, 0.06 mmol) as a white solid. 1H NMR(500MHz,DMSO-d6+K2CO3)ppm 7.94-7.99(m,2H),7.82(t,1H),7.67(dd,1H),7.07(dd,1H),6.89(dd,1H),4.16(s ,2H),3.86(d,2H),3.78(d,2H),3.39-3.46(m,4H),2.51(s,2H),2.23-2.32(m,4H). LCMS[M+H] + 514.1,516.1.

[0440] Example 51-Compound 51

[0441]

[0442] Tert-butyl (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)carbamate was prepared according to Reaction Scheme 20:

[0443]

[0444] Reagents: Step 1) diphenylphosphoryl azide, Et3N, toluene, 100°C; tert-butyl alcohol, 100°C.

[0445] Step 1. To a suspension of 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (124 mg, 0.27 mmol) in toluene (4 mL) was added neat Et3N (50 μL, 0.36 mmol) and diphenylphosphoryl azide (70 μL, 0.33 mmol), and the mixture was stirred at 100°C under N2 for 17 h. The reaction mixture was cooled to rt, tert-butanol (2 mL) was added, and it was heated to 100°C for 4 h. The reaction mixture was cooled, concentrated in vacuo to remove most of the toluene and tert-butanol, transferred into EtOAc (15 mL), washed with saturated aqueous NaHCO3 (2×15 mL) and brine (15 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexanes / EtOAc) to give the title compound as a colorless glass (33.8 mg). 1H NMR(500MHz, CDCl3)δppm7.97(d,1H),7.51-7.58(m,2H),7.39(dd,1H),6.77(dd,1H) ,6.73(dd,1H),4.98(br.s,1H),4.29(s,2H),4.21(d,2H),4.08(d,2H),1.42(s,9H). LCMS[M+H] + 530.1,531.8.

[0446] Example 52-Compound 52

[0447]

[0448] 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride was prepared according to Reaction Scheme 21:

[0449]

[0450] Reagents: Step 1) LHMDS, THF, –78°C; CBr4; 2) NaN3, DMSO; 3) H2, Pd / C, EtOH; 4) Benzyl chloroformate, saturated aqueous NaHCO3, di alkane; 5) LiAlH(Ot-Bu)3, THF; 6) methanesulfonyl chloride, Et3N, DCM; 7) 2-fluoro-4-hydroxybenzonitrile, K2CO3, CH3CN, Δ; 8) TFA, DCM; 9) 2,4-dichlorobenzenesulfonyl chloride, Et3N, DCM; 10) TFA, 50°C.

[0451] Step 1. 3-bromoazetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-ethyl ester. A mixture of LHMDS (1.0 M in THF / ethylbenzene, 15.8 mL, 15.8 mmol) and anhydrous THF (60 mL) was cooled to -78 °C under N2. A solution of 1-boc-azetidine-3-carboxylic acid ethyl ester (3.01 g, 13.1 mmol) in anhydrous THF (30 mL) was added dropwise, and the mixture was stirred at -78 °C for 15 min. A solution of CBr4 (6.56 g, 19.8 mmol) in anhydrous THF (30 mL) was added dropwise, the mixture was warmed to rt and stirred for 20 h. The reaction mixture was diluted with Et2O (350 mL), washed with saturated aqueous NaHCO3 (2×250 mL) and brine (250 mL), dried over Na2SO4, decanted and concentrated. The residue was purified by flash chromatography (EtOAc / hexanes) to give the title compound as an orange oil (2.53 g). 1H NMR (500MHz, CDCl3) δppm 4.64 (dd, 2H), 4.25-4.33 (m, 4H), 1.45 (s, 9H), 1.33 (t, 3H).

[0452] Step 2. 3-Azidoazetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-ethyl ester. A mixture of 3-bromoazetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-ethyl ester (2.53 g, 8.2 mmol) and NaN3 (1.59 g, 24.5 mmol) in anhydrous DMSO (20 mL) was stirred at 50°C under N2 for 18 h. The reaction mixture was cooled, diluted with Et2O (250 mL), washed with saturated aqueous NaHCO3 solution (150 mL) and brine (2×150 mL), dried over Na2SO4, filtered and concentrated to dryness to give the title compound (2.10 g) as a yellow oil. 1 H NMR (500MHz, CDCl3) δppm 4.30-4.36 (m, 4H), 3.97 (d, 2H), 1.45 (s, 9H), 1.36 (t, 3H).

[0453] Step 3. 3-aminoazetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-ethyl ester. A mixture of 3-azidoazetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-ethyl ester (1.93 g, 7.15 mmol) and Pd / C (300 mg, 10 wt%, 50% H2O, Degussa) in EtOH (70 mL) was stirred under H2 atmosphere for 16 h. The reaction mixture was filtered through celite and the filtrate was concentrated to dryness to give the title compound (1.73 g) as a yellow oil. 1 H NMR (500MHz, CDCl3) δppm 4.22-4.32(m,4H),3.79(d,2H),2.37(br.s,2H),1.44(s,9H),1.31(t,3H).

[0454] Step 4. 1-(tert-butyl)3-ethyl 3-(((benzyloxy)carbonyl)amino)azetidine-1,3-dicarboxylate. 1-(tert-butyl)3-ethyl 3-aminoazetidine-1,3-dicarboxylate (1.89 g, 7.72 mmol) and benzyl chloroformate (1.76 mL, 12.4 mmol) were dissolved in saturated aqueous NaHCO3 (40 mL) and distilled water. To the mixture of 4-nitro-1-yl pyridine (2-nitro-2-yl)-2-nitropropene (40mL) under N2 and stirred vigorously for 4 days.The reaction mixture is diluted with EtOAc (250mL), washed with saturated NaHCO3 aqueous solution (200mL) and salt solution (200mL), at Na2SO4 upper drying, decantation and concentration.Residue is purified by flash chromatography (EtOAc / hexane), obtain the title compound (2.49g) as colorless oil, which is partially solidified. 1 H NMR (500MHz, CDCl3) δppm7.29-7.41(m,5H),5.58(br.s,1H),5.13(s,2H),4.22-4.39(m,4H),4.07-4.22(m,2H),1.45(s,9H),1.23-1.34(m,3H).

[0455] Step 5. tert-Butyl 3-(((benzyloxy)carbonyl)amino)-3-(hydroxymethyl)azetidine-1-carboxylate. To a solution of 1-(tert-butyl) 3-ethyl 3-(((benzyloxy)carbonyl)amino)azetidine-1,3-dicarboxylate (2.31 g, 6.11 mmol) in anhydrous THF was added LiAlH(Ot-Bu)3 (1.0 M in THF, 15.3 mL) dropwise at rt under N2 and the mixture was stirred for 22 h. The reaction mixture was diluted with EtOAc (250 mL), washed with 1 M HCl (2×150 mL) and brine (150 mL), dried over Na2SO4, filtered and concentrated to dryness to give the title compound (1.89 g) as a white solid. 1 H NMR (500MHz, CDCl3) δppm7.31-7.40(m,5H),5.34(br.s,1H),5.09(s,2H),3.93-4.02(m,2H),3.82-3.92(m,4H),2.63(br.s,1H),1.43(s,9H).

[0456] Step 6. tert-Butyl 3-(((benzyloxy)carbonyl)amino)-3-(((methylsulfonyl)oxy)methyl)azetidine-1-carboxylate. To a solution of tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-(hydroxymethyl)azetidine-1-carboxylate (1.94 g, 5.8 mmol) in anhydrous DCM (50 mL) was added EtN (1.2 mL, 8.6 mmol) and methanesulfonyl chloride (0.53 mL, 6.8 mmol). The mixture was stirred at rt under N for 3 h, poured into saturated aqueous NaHCO (100 mL) and extracted with DCM (2×100 mL). The extract was dried over NaSO, filtered and concentrated to dryness to give the title compound (2.36 g) as a light orange foam.1 H NMR (500MHz, CDCl3) δppm 7.29-7.44(m,5H),5.41(br s,1H),5.10(s,2H),4.50(s,2H),3.88-4.00(m,4H),2.95(s,3H),1.43(s,9H).

[0457] Step 7. tert-Butyl 3-(((benzyloxy)carbonyl)amino)-3-((4-cyano-3-fluorophenoxy)methyl)azetidine-1-carboxylate. A mixture of tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-(((methylsulfonyl)oxy)methyl)azetidine-1-carboxylate (2.36 g, 5.7 mmol), 2-fluoro-4-hydroxybenzonitrile (936 mg, 6.8 mmol) and KCO (1.59 g, 11.5 mmol) in anhydrous CHCN (50 mL) was stirred at reflux under N for 16 h. The reaction mixture was cooled, diluted with EtOAc (250 mL), washed with 2M KCO (4×150 mL) and brine (150 mL), dried over NaSO, decanted and concentrated. The residue was purified by flash chromatography (EtOAc / hexanes) to give the title compound as a foam (1.81 g). 1 H NMR(500MHz,CDCl3)δppm 7.51(dd,1H),7.29-7.39(m,5H),6.66-6.81(m,2H),5.30(br.s,1H),5.08(s,2H),4.29(m,2H),3.95-4.08(m,4H),1.44(s,9H).

[0458] Step 8. Benzyl (3-((4-cyano-3-fluorophenoxy)methyl)azetidine-3-yl)carbamate trifluoroacetate. A mixture of tert-butyl 3-(((benzyloxy)carbonyl)amino)-3-((4-cyano-3-fluorophenoxy)methyl)azetidine-1-carboxylate (1.81 g, 3.97 mmol) and TFA (10 mL) in anhydrous DCM (40 mL) was stirred at rt under N2 for 2 h. The reaction mixture was concentrated to dryness to give the crude title compound (2.21 g) as a foam.

[0459] Step 9. Benzyl (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)carbamate. A mixture of benzyl (3-((4-cyano-3-fluorophenoxy)methyl)azetidin-3-yl)carbamate trifluoroacetate (2.21 g, 3.97 mmol), Et3N (1.70 mL, 12.2 mmol) and 2,4-dichlorobenzenesulfonyl chloride (1.17 g, 4.77 mmol) in anhydrous DCM (40 mL) was stirred at rt under N2 for 18 h. The reaction mixture was poured into saturated aqueous NaHCO3 (100 mL) and extracted with DCM (2×100 mL). The extracts were dried over Na2SO4, decanted and concentrated, and the residue was purified by flash chromatography (EtOAc / hexanes) to give the title compound as a foam (2.07 g). 1 H NMR (500MHz, CDCl3) δppm 7.97(d,1H),7.56(d,1H),7.52(dd,1H),7.38(dd,1H),7.28-7.36(m,5H),6.65-6.78(m,2H),5.26(br s,1H),5.07(s,2H),4.30(br s,2H),4.24(d,2H),4.11(d,2H).

[0460] Step 10. 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride. A mixture of benzyl (3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)carbamate (883 mg, 1.56 mmol) and TFA (12 mL) was stirred at 50 °C for 7 h. The reaction mixture was concentrated, transferred to DCM, poured into saturated aqueous NaHCO3 (100 mL), and extracted with DCM (3×75 mL). The extract was dried over Na2SO4, decanted and concentrated, and the residue was purified twice by flash chromatography (DCM / CH3OH / NH4OH and hexane / EtOAc / CH3OH). The pure product was purified by distillation. The resulting product was treated with 4 M HCl in dextran, concentrated, and triturated with CH 3 OH / Et 2 O. The precipitate was collected by vacuum filtration to give the title compound (216 mg) as a white solid. 1H NMR(500MHz,DMSO-d6)δppm 9.05(br s,3H),7.93-8.01(m,2H),7.87(t,1H),7.65(dd,1H),7.12(dd,1H),6.96(dd,1H),4.43(s,2H),4.23(d,2H),4.04(d,2H). LCMS[M+H] + 430.0,432.0.

[0461] Example 53-Compound 53

[0462]

[0463] N-(3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)acetamide was prepared as described below. To a suspension of 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride (10.4 mg, 0.022 mmol) in anhydrous DCM (0.5 mL) was added neat triethylamine (10 μL, 0.072 mmol) and acetyl chloride (0.33 mL, 0.1 M in DCM), and the mixture was stirred at rt for 24 h. Additional triethylamine (40 μL, 0.287 mmol) and acetyl chloride (16 μL, 0.225 mmol) were added and stirring was continued for another 18 h. The mixture was quenched with CH3OH (0.5 mL), diluted with DCM (5 mL), dry loaded onto celite, and purified by flash chromatography (EtOAc / hexanes) to give the title compound as a light yellow solid (7.2 mg). 1 H NMR (500MHz, CDCl3) δppm7.97(d,1H),7.51-7.58(m,2H),7.39(dd,1H),6.75(dd,1 H),6.70(dd,1H),5.90(s,1H),4.34(s,2H),4.20(d,2H),4.14(d,2H),2.00(s,3H). LCMS[M+H] + 472.0,474.1.

[0464] Example 54-Compound 54

[0465]

[0466] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(dimethylamino)azetidine-3-yl)methoxy)-2-fluorobenzonitrile was prepared as described below. A mixture of 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride (10.8 mg, 0.023 mmol) and formaldehyde (37% aqueous solution, 1.9 μL, 0.03 mmol) in anhydrous THF (0.25 mL) was stirred at rt for 15 min. NaBH(OAc)3 (21 mg, 0.1 mmol) was added, and the mixture was stirred at rt for 23 h. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (1 mL) and extracted with EtOAc (2×1 mL). The extract was dried over Na2SO4, decanted and concentrated in vacuo. The residue was purified by flash chromatography (hexanes / EtOAc / CH3OH) to give the title compound (5.1 mg) as a white solid (-80% purity). 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.52-7.58(m,2H),7.38(dd,1H),6.79(dd,1H),6.74(dd,1H),4.24(s,2H),4.03(d,2H),3.91(d,2H),2.31(s,6H). LCMS[M+H] + 458.1,460.1.

[0467] Example 55-Compound 55

[0468]

[0469] 2-Amino-N-(3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)acetamide was prepared according to Reaction Scheme 22:

[0470]

[0471] Reagents: Step 1) Boc-glycine, HATU, DIPEA, DMF; 2) Di HCl in alkanes.

[0472] Step 1. tert-Butyl (2-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)amino)-2-ketoethyl)carbamate. A mixture of 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride (25 mg, 0.053 mmol), Boc-glycine (12 mg, 0.067 mmol), HATU (24 mg, 0.063 mmol) and DIPEA (28 μL, 0.16 mmol) in anhydrous DMF (0.4 mL) was stirred at rt for 17 h. The reaction mixture was diluted with EtOAc (10 mL), washed with saturated aqueous NaHCO3 (2×7 mL) and brine (7 mL), dried over Na2SO4, decanted and concentrated. The residue was purified by flash chromatography (EtOAc / hexanes) to provide the title compound.

[0473] Step 2. 2-amino-N-(3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)acetamide. Tert-butyl (2-((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)amino)-2-ketoethyl)carbamate (0.053 mmol) and dihydrochloride were added. A mixture of 4.0 M HCl in oxane (1.0 mL) was stirred at rt for 18 hr and concentrated to dryness. The residue was purified by flash chromatography (DCM / CH3OH / NH4OH) to give the title compound (9.8 mg) as a white solid. 1 H NMR (500MHz, CDCl3) δppm7.93-8.01(m,2H),7.49-7.59(m,2H),7.39(dd,1H), 6.67-6.78(m,2H),4.34(s,2H),4.27(d,2H),4.18(d,2H),3.30-3.39(m,2H). LCMS[M+H] + 487.1,489.0.

[0474] Example 56-Compound 56

[0475]

[0476] N-(3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)-2-hydroxyacetamide was prepared in a manner similar to that in Scheme 22 from 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride and glycolic acid. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.57(d,1H),7.54(dd,1H),7.39(dd,1H),7.03(s,1H),6.76(dd,1H) ,6.71(dd,1H),4.36(s,2H),4.28(d,2H),4.19(d,2H),4.13(s,2H),2.81(s,1H). LCMS[M+H] + 488.1,490.1.

[0477] Example 57-Compound 57

[0478]

[0479] 2-Amino-N-(3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)ethane-1-sulfonamide hydrochloride was prepared in a similar manner to that in Scheme 13 from 4-((3-amino-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile hydrochloride and Boc-taurine chloride. 1 H NMR(500MHz,DMSO-d6)δppm 8.58(s,1H),8.05(br s,3H),7.94-7.99(m,2H),7.86(t,1H),7.67(dd,1H),7.17(dd,1H),6.97(dd,1H ),4.34(s,2H),4.16(d,2H),4.01(d,2H),3.36-3.42(m,2H),3.08-3.17(m,2H). LCMS[M+H] + 537.1,538.9.

[0480] Example 58-Compound 58

[0481]

[0482] 4-((1-((2-chlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared according to Reaction Scheme 23:

[0483]

[0484] Reagents: Step 1) di-tert-butyl dicarbonate, saturated aqueous NaHCO3 solution, di alkane; 2) LiAlH(Ot-Bu)3, THF; 3) methanesulfonyl chloride, Et3N, DCM; 4) 2-fluoro-4-hydroxybenzonitrile, K2CO3, CH3CN, Δ; 5) LiAlH(Ot-Bu)3, THF; 6) TFA, DCM; 7) 2-chlorobenzenesulfonyl chloride, Et3N, DCM.

[0485] Step 1. 1-(tert-butyl)3,3-diethyl azetidine-1,3,3-tricarboxylate. Azetidine-3,3-dicarboxylic acid diethyl ester hydrochloride (3.46 g, 14.55 mmol), di-tert-butyl dicarbonate (4.7 mL, 20.5 mmol), saturated aqueous NaHCO3 solution (75 mL) and distilled water were stirred for 2 hours. The mixture of 1,2-dihydro-1,4-dihydro-2-nitropropane (75mL) was stirred vigorously at rt for 18h. The reaction mixture was diluted with saturated NaHCO3 aqueous solution (100mL) and water and extracted with EtOAc (3×100mL). The extract was washed with brine (200mL), dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica gel (hexane / EtOAc) to give the title compound (4.02g) as a colorless oil. 1 H NMR (500MHz, CDCl3) δppm4.23-4.29(m,8H),1.44(s,9H),1.28(t,6H).

[0486] Step 2.3-(Hydroxymethyl)azetidine-1,3-dicarboxylic acid 1-(tert-butyl) 3-ethyl ester. At 0 ° C and N2, a solution of azetidine-1,3,3-tricarboxylic acid 1-(tert-butyl) 3,3-diethyl ester (4.02g, 13.3mmol) in anhydrous THF (120mL) was added dropwise by an addition funnel. LiAlH(Ot-Bu)3(1.0M in THF, 27.5mL) was added dropwise, and the mixture was warmed to rt and stirred for 18h. The reaction mixture was diluted with EtOAc (150mL) and washed with 1M HCl (200mL). The aqueous layer was extracted with EtOAc (150mL), and the combined organic phases were washed with brine (200mL), dried over Na2SO4, filtered and concentrated to give the title compound (3.85g) as a colorless oil. 1 H NMR (500MHz, CDCl3) δppm 4.26 (q, 2H), 4.15 (d, 2H), 3.95 (s, 2H), 3.81 (d, 2H), 1.45 (s, 9H), 1.32 (t, 3H).

[0487] Step 3. 1-(tert-butyl)3-ethyl 3-(((methylsulfonyl)oxy)methyl)azetidine-1,3-dicarboxylate. To a solution of 1-(tert-butyl)3-ethyl 3-(hydroxymethyl)azetidine-1,3-dicarboxylate (460 mg, 1.774 mmol) and Et3N (0.37 mL, 2.654 mmol) in anhydrous DCM (12 mL) was added methanesulfonyl chloride (0.17 mL, 2.2 mmol) dropwise, and the mixture was stirred at rt for 2.5 h. The reaction mixture was poured into saturated aqueous NaHCO3 (75 mL) and extracted with DCM (3×35 mL). The extract was dried over Na2SO4, filtered and concentrated in vacuo to give the crude title compound (679 mg) as a colorless oil.

[0488] Step 4. 1-(tert-butyl)3-ethyl 3-((4-cyano-3-fluorophenoxy)methyl)azetidine-1,3-dicarboxylate. A mixture of crude 1-(tert-butyl)3-ethyl 3-(((methylsulfonyl)oxy)methyl)azetidine-1,3-dicarboxylate (679 mg, 2.2 mmol), 2-fluoro-4-hydroxybenzonitrile (294 mg, 2.14 mmol) and K2CO3 (493 mg, 3.6 mmol) in anhydrous DMF (10 mL) was stirred at 85 °C under N2 for 18 h. The reaction mixture was cooled, diluted with Et2O (100 mL), washed with 2M K2CO3 (2×75 mL) and brine (75 mL), dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica gel (hexanes / EtOAc) to give the title compound (590 mg, 1.56 mmol) as a viscous colorless oil. 1 H NMR (500MHz, CDCl3) δppm 7.54(dd,1H),6.78(dd,1H),6.74(dd,1H),4.35(s,2H),4.22-4.30(m,4H),3.93(d,2H),1.45(s,9H),1.28(t,3H).

[0489] Step 5. tert-butyl 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-carboxylate. To a solution of 1-(tert-butyl) 3-ethyl 3-((4-cyano-3-fluorophenoxy)methyl)azetidine-1,3-dicarboxylate (590 mg, 1.56 mmol) in anhydrous THF (12 mL) was added dropwise a solution of LiAlH(Ot-Bu)3 (1.0 M in THF, 4.4 mL) at 0 °C under N2. The mixture was warmed to rt and stirred for 4 days. The reaction mixture was diluted with EtOAc (50 mL), washed with 1 M HCl (2×50 mL) and brine (50 mL), dried over Na2SO4, filtered and concentrated to dryness to give the title compound (523 mg) as a viscous colorless oil. 1 H NMR (500MHz, CDCl3) δppm 7.54 (dd, 1H), 6.79 (dd, 1H), 6.75 (dd, 1H), 4.18 (s, 2H), 3.91 (s, 2H), 3.77-3.83 (m, 4H), 1.45 (s, 9H).

[0490] Step 6. 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate. A mixture of tert-butyl 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-carboxylate (523 mg, 1.555 mmol) and TFA (1.5 mL) in anhydrous DCM (15 mL) was stirred at rt under N for 3 h. The reaction mixture was concentrated to dryness and concentrated again from DCE, toluene and CHOH and finally from EtO to give the title compound (577 mg) as a viscous colorless oil which slowly solidified.

[0491] Step 7. 4-((1-((2-chlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile. To a solution of 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate (19.6 mg, 0.056 mmol) and Et3N (31 μL, 0.222 mmol) in anhydrous DCM (0.5 mL) was added 2-chlorobenzenesulfonyl chloride (12.0 mg, 0.057 mmol) and the mixture was stirred at rt for 6 h. The mixture was concentrated and purified by flash chromatography on silica gel (hexane / DCM / CH3CN) to give the title compound (13.1 mg) as a colorless glass. 1H NMR(500MHz,CDCl3)δppm 8.05(dd,1H),7.49-7.59(m,3H),7.38-7.44(m,1H),6.75(dd,1H),6.69(dd,1H),4.16(s,2H),3.97(d,2H),3.93(d,2H),3.90(s,2H),1.72(br s,1H). LCMS[M+H] + 411,413.

[0492] Example 59-Compound 59

[0493]

[0494] 4-((1-((2-chloro-4-(trifluoromethyl)phenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-chloro-4-(trifluoromethyl)benzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.18(d,1H),7.81(d,1H),7.66(dd,1H),7.54(dd,1H),6.69-6.78(m,2H),4.17(s,2H),3.97-4.06(m,4H),3.92(s,2H),1.71(br s,1H). LCMS[M+H] + 479,481.

[0495] Example 60-Compound 60

[0496]

[0497] 4-((1-((2-chloro-4-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-chloro-4-cyanobenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm8.16(d,1H),7.83(d,1H),7.69(dd,1H),7.55(dd,1H),6.79(dd ,1H),6.74(dd,1H),4.18(s,2H),4.05(d,2H),4.01(d,2H),3.91(s,2H),1.70(br.s,1H). LCMS[M+H]+ 436.

[0498] Example 61-Compound 61

[0499]

[0500] 4-((1-((3,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 3,4-dichlorobenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.93-7.97(m,1H),7.65-7.72(m,2H),7.48-7.55(m,1H),6.55-6.63(m,2H),4.04(s,2H),3.79(s,2H),3.72-3.78(m,4H),1.70(br s,1H). LCMS[M+H] + 445,447.

[0501] Example 62-Compound 62

[0502]

[0503] 4-((1-((2,4-dimethylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2,4-dimethylbenzenesulfonyl chloride. 1 H NMR(500MHz, CDCl3)δppm7.82(d,1H),7.51(dd,1H),7.10-7.17(m,2H),6.72(dd,1H),6.64(dd, 1H),4.11(s,2H),3.88(d,2H),3.77(d,2H),3.73(d,2H),2.61(s,3H),2.39(s,3H),1.69(t,1H). LCMS[M+H] + 405.

[0504] Example 63-Compound 63

[0505]

[0506] Methyl 4-((3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidin-1-yl)sulfonyl)benzoate was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and methyl 4-(chlorosulfonyl)benzoate. 1 H NMR (500MHz, CDCl3) δppm8.25(d,2H),7.93(d,2H),7.48(dd,1H),6.58(dd,1 H),6.48(dd,1H),3.99(s,3H),3.98(s,2H),3.68-3.77(m,6H),1.61(t,1H). LCMS[M+H] + 435.

[0507] Example 64-Compound 64

[0508]

[0509] 2-Fluoro-4-((3-(hydroxymethyl)-1-((6-(trifluoromethyl)pyridin-3-yl)sulfonyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 6-(trifluoromethyl)pyridine-3-sulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 9.18(d,1H),8.34(dd,1H),7.92(d,1H),7.47-7.56(m,1H),6.56-6.66(m,2H),4.06(s,2H),3.80-3.88(m,4H),3.78(d,2H),1.62(t,1H). LCMS[M+H] + 446.

[0510] Example 65-Compound 65

[0511]

[0512] 4-((1-((2,4-dimethylthiazol-5-yl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2,4-dimethylthiazole-5-sulfonyl chloride. 1H NMR(500MHz,CDCl3)δppm 7.53(dd,1H),6.69(dd,1H),6.63(dd,1H),4.08(s,2H),3.81-3.87(m,4H),3.75-3.80(m,2H),2.73(s,3H),2.67(s,3H),1.69(t,1H). LCMS[M+H] + 412.

[0513] Example 66-Compound 66

[0514]

[0515] 4-((1-((1,3-dimethyl-1H-pyrazol-4-yl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 1,3-dimethyl-1H-pyrazole-4-sulfonyl chloride. 1 H NMR(500MHz, CDCl3)δppm7.76(s,1H),7.53(dd,1H),6.70(dd,1H),6.65(dd,1H) ,4.08(s,2H),3.89(s,3H),3.85(s,2H),3.72(d,2H),3.67(d,2H),2.45(s,3H). LCMS[M+H] + 395.

[0516] Example 67-Compound 67

[0517]

[0518] 4-((1-((3-chloro-4-fluorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 3-chloro-4-fluorobenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.95(dd,1H),7.74-7.80(m,1H),7.52(dd,1H),7.36(t,1H),6.60-6.68(m,2H),4.07(s,2H),3.71-3.80(m,6H),1.66(t,1H). LCMS[M+H] + 429,431.

[0519] Example 68-Compound 68

[0520]

[0521] 4-((1-((3,5-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 3,5-dichlorobenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm 7.74(d,2H),7.63-7.66(m,1H),7.53(dd,1H),6.67(dd,1H),6.62(dd,1H),4.07(s,2H),3.74-3.83(m,6H),1.60(t,1H). LCMS[M+H] + 445,447.

[0522] Example 69-Compound 69

[0523]

[0524] 2-Fluoro-4-((3-(hydroxymethyl)-1-((4-(methylsulfonyl)phenyl)sulfonyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 4-(methylsulfonyl)benzenesulfonyl chloride. 1 H NMR(500MHz,DMSO-d6)δppm 8.19-8.25(m,2H),8.06-8.11(m,2H),7.69-7.75(m,1H),6.97(dd,1H),6.6 2(dd,1H),4.98(t,1H),3.94(s,2H),3.64(s,4H),3.37(d,2H),3.32(s,3H). LCMS[M+H] + 455.

[0525] Example 70-Compound 70

[0526]

[0527] 2-Fluoro-4-((3-(hydroxymethyl)-1-(((3-(trifluoromethyl)phenyl)sulfonyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 3-(trifluoromethyl)benzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.14(s,1H),8.07(d,1H),7.94(d,1H),7.76(t,1H),7.51(dd,1H),6.62(dd,1H),6.56(dd,1H),4.04(s,2H),3.72-3.81(m,6H),1.57(t,1H). LCMS[M+H] + 445.

[0528] Example 71-Compound 71

[0529]

[0530] 4-((1-((2-chloropyridin-3-yl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-chloropyridine-3-sulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.58(dd,1H),8.36(dd,1H),7.54(dd,1H),7.42(dd,1H),6.78(dd,1H),6.7 3(dd,1H),4.18(s,2H),4.06(d,2H),4.02(d,2H),3.93(d,2H),1.76(t,1H). LCMS[M+H] + 453,455.

[0531] Example 72-Compound 72

[0532]

[0533] 2-Fluoro-4-((3-(hydroxymethyl)-1-(phenylsulfonyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and benzenesulfonyl chloride. 1HNMR(500MHz,CDCl3)δppm 7.85-7.90(m,2H),7.68-7.74(m,1H),7.59-7.65(m,2H),7.49(dd,1H),6.59(dd,1 H),6.52(dd,1H),3.97(s,2H),3.71-3.77(m,4H),3.66-3.71(m,2H),1.58(t,1H). LCMS[M+H] + 377.

[0534] Example 73-Compound 73

[0535]

[0536] 2-Fluoro-4-((3-(hydroxymethyl)-1-tosylazetidin-3-yl)methoxy)benzonitrile was prepared in a similar manner to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 4-methyl-benzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.73-7.79(m,2H),7.50(dd,1H),7.41(d,2H),6.59(dd,1H),6.46(dd,1H),3 .95(s,2H),3.77(d,2H),3.71(d,2H),3.66(d,2H),2.50(s,3H),1.57(t,1H). LCMS[M+H] + 391.

[0537] Example 74-Compound 74

[0538]

[0539] 4-((1-((4-chloro-2-methylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-methyl-4-chlorobenzenesulfonyl chloride. 1H NMR (500MHz, CDCl3) δppm7.89(d,1H),7.54(dd,1H),7.35(d,1H),7.31(dd,1H),6.73(dd,1H),6. 70(dd,1H),4.14(s,2H),3.90(s,2H),3.80-3.84(m,2H),3.75-3.79(m,2H),2.63(s,3H),1.67(br s,1H).

[0540] Example 75-Compound 75

[0541]

[0542] 4-((1-((2,6-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a similar manner to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2,6-dichlorobenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.53(dd,1H),7.48(d,2H),7.32-7.38(m,1H),6.77(dd,1H),6.71(dd,1H) ,4.18(s,2H),4.04-4.09(m,2H),3.99-4.03(m,2H),3.93(s,2H),1.71(br s,1H).

[0543] Example 76-Compound 76

[0544]

[0545] 2-Fluoro-4-((3-(hydroxymethyl)-1-(o-toluenesulfonyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-methyl-benzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.92-7.98(m,1H),7.46-7.55(m,2H),7.31-7.38(m,2H),6.73(dd,1H),6.68(dd,1H), 4.13(s,2H),3.89(s,2H),3.79-3.83(m,2H),3.75-3.78(m,2H),2.65(s,3H),1.82(br s,1H).

[0546] Example 77-Compound 77

[0547]

[0548] 2-Fluoro-4-((1-((4-fluoro-2-methylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-methyl-4-fluorobenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.97(dd,1H),7.53(dd,1H),6.97-7.08(m,2H),6.75(dd,1H),6.71(dd,1H),4.16(s,2 H),3.90(s,2H),3.79-3.84(m,2H),3.74-3.79(m,2H),2.65(s,3H),1.62-1.73(m,1H).

[0549] Example 78-Compound 78

[0550]

[0551] 4-((1-((4-chloro-2-fluorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-fluoro-4-chlorobenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm7.79-7.86(m,1H),7.52(dd,1H),7.28-7.36(m,2H),6. 67(dd,1H),6.63(dd,1H),4.10(s,2H),3.85-3.91(m,4H),3.84(s,2H),1.78(br s,1H).

[0552] Example 79-Compound 79

[0553]

[0554] 4-((1-((4-cyano-2-methylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-methyl-4-cyanobenzenesulfonyl chloride.1 H NMR(500MHz,CDCl3)δppm 8.06(d,1H),7.59-7.66(m,2H),7.49-7.57(m,1H),6.77(dd,1H),6.73(dd,1 H),4.17(s,2H),3.87-3.94(m,4H),3.82-3.87(m,2H),2.68(s,3H),1.84(br s,1H).

[0555] Example 80-Compound 80

[0556]

[0557] Methyl 3-chloro-4-((3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidin-1-yl)sulfonyl)benzoate was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and methyl 3-chloro-4-(chlorosulfonyl)benzoate. 1 H NMR(500MHz,CDCl3)δppm 8.19(d,1H),8.11(d,1H),8.03(dd,1H),7.52(dd,1H),6.75(dd,1H),6.69 (dd,1H),4.16(s,2H),3.94-4.03(m,7H),3.90(s,2H),1.64-1.84(m,1H).

[0558] Example 81-Compound 81

[0559]

[0560] 4-((1-((4-chloro-2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-chloro-benzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm8.01(d,1H),7.85(d,1H),7.71(dd,1H),7.54(dd,1H),6.76( dd,1H),6.71(dd,1H),4.17(s,2H),3.96-4.05(m,4H),3.90(s,2H),1.67-1.76(m,1H). LC / MS / MS[MH] - 479.8.

[0561] Example 82-Compound 82

[0562]

[0563] 4-((1-((2-chloro-4-methylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-chloro-4-methylbenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm7.91(d,1H),7.52(t,1H),7.37(s,1H),7.20(d,1H),6. 73(dd,1H),6.65(dd,1H),4.14(s,2H),3.83-3.99(m,6H),2.42(s,3H),1.74(br s,1H).

[0564] Example 83-Compound 83

[0565]

[0566] 4-((1-((2-chloro-4-methoxyphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-chloro-4-methoxybenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm 7.96(d,1H),7.52(dd,1H),7.06(d,1H),6.88(dd,1H),6.74(dd,1H),6.67(dd,1H),4.14(s,2H),3.82-3.94(m,9H),1.81(br s,1H).

[0567] Example 84-Compound 84

[0568]

[0569] 4-((1-((4-chlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 4-chlorobenzenesulfonyl chloride. 1H NMR(500MHz,CDCl3)δppm 7.79-7.84(m,2H),7.57-7.62(m,2H),7.51(t,1H),6.53-6.61(m,2H),3.99(s,2H),3.77(s,2H),3.66-3.75(m,4H),1.66(br s,1H).

[0570] Example 85-Compound 85

[0571]

[0572] 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyanobenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.09(dd,1H),7.90(dd,1H),7.69-7.80(m,2H),7.53(dd,1H),6.66-6.79(m,2H),4.17(s,2H),3.97-4.04(m,4H),3.90(d,2H),1.75(t,1H).

[0573] Example 86-Compound 86

[0574]

[0575] 4-((1-((2-cyano-4-(trifluoromethyl)phenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-trifluoromethylbenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.23(d,1H),8.13(d,1H),7.98-8.02(m,1H),7.53(dd,1H),6.76(dd,1H) ,6.71-6.74(m,1H),4.18(s,2H),4.02-4.10(m,4H),3.91(s,2H),1.84(br s,1H).

[0576] Example 87-Compound 87

[0577]

[0578] (1-((2,4-dichlorophenyl)sulfonyl)-3-((3,4-difluorophenoxy)methyl)azetidin-3-yl)methanol was prepared according to Reaction Scheme 24:

[0579]

[0580] Reagents: Step 1) TBS-Cl, imidazole, DMF; 2) TFA, DCM; 3) 2,4-dichlorobenzenesulfonyl chloride, Et3N, DCM; 4) LiAlH(Ot-Bu)3, THF; 5) methanesulfonyl chloride, Et3N, DCM; 6) 3,4-difluorophenol, K2CO3, DMF, 85°C; 7) TBAF, THF.

[0581] Step 1. 1-(tert-butyl)3-ethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)azetidine-1,3-dicarboxylate. A mixture of 1-(tert-butyl)3-ethyl 3-(hydroxymethyl)azetidine-1,3-dicarboxylate (3.85 g 13.3 mmol), tert-butyldimethylsilyl chloride (2.48 g, 16.46 mmol) and imidazole (1.84 g, 27.1 mmol) in anhydrous DMF (50 mL) was stirred at rt under N2 for 20 h. More tert-butyldimethylsilyl chloride (1.38 g, 9.17 mmol) and imidazole (0.95 g, 14 mmol) were added and stirring was continued for 3 days. The reaction mixture was diluted with EtOAc (300 mL), washed with 1M HCl (2 x 150 mL) and brine (150 mL), dried over NaSO, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica gel (hexane / EtOAc) to give the title compound (4.84 g) as a colorless oil. 1 HNMR (500MHz, CDCl3) δppm 4.20(q,2H),4.10(d,2H),3.88(s,2H),3.83(d,2H),1.43(s,9H),1.28(t,3H),0.87(s,9H),0.05(s,6H).

[0582] Step 2. Ethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)azetidine-3-carboxylate trifluoroacetate. A mixture of 1-(tert-butyl)3-ethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)azetidine-1,3-dicarboxylate (832 mg, 2.00 mmol) and TFA (2.0 mL) in anhydrous DCM (20 mL) was stirred at rt under N2 for 2 h. The reaction mixture was concentrated to dryness, concentrated twice from toluene and once from Et2O to give the title compound (840 mg) which was used directly as a crude product.

[0583] Step 3. Ethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylate. To a solution of ethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)azetidine-3-carboxylate trifluoroacetate (840 mg, theoretical value 2.00 mmol) and Et3N (0.70 mL, 5.02 mmol) in anhydrous DCM (20 mL) was added 2,4-dichlorobenzenesulfonyl chloride (467 mg, 1.9 mmol) and the mixture was stirred at rt under N2 for 20 h. The reaction mixture was poured into saturated aqueous NaHCO3 (50 mL) and extracted with DCM (3×35 mL). The extract was dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica gel (hexanes / EtOAc) to give the desilylated by-product as a colorless oil (187 mg) and the title compound as a colorless oil (617 mg). 1 HNMR(500MHz,CDCl3)δppm 7.96(d,1H),7.54(d,1H),7.36(dd,1H),4.24(d,2H),4.19(q,2H),3.99 (d,2H),3.89(s,2H),1.26(t,3H),0.82-0.87(m,9H),0.02-0.06(m,6H).

[0584] Step 4. (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-yl)methanol. To a solution of ethyl 3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylate (809 mg, 1.68 mmol) in anhydrous THF (12 mL) was added dropwise a solution of LiAlH(Ot-Bu)3 (1 M in THF, 5 mL) at 0°C under N2. The mixture was warmed to rt and stirred for 20 h. Another portion of LiAlH(Ot-Bu)3 (1 M in THF, 1.7 mL) was added dropwise and stirred for 22 h. The reaction mixture was diluted with EtOAc (100 mL), washed with 1M HCl (2 x 75 mL) and brine (75 mL), dried over Na2SO4, filtered and concentrated to give the title compound as a colorless oil (720 mg). 1H NMR (500MHz, CDCl3) δppm 7.97(d,1H),7.55(d,1H),7.37(dd,1H),3.74-3.83(m,8H),0.85-0.90(m,9H),0.05-0.09(m,6H).

[0585] Step 5. (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate. To a solution of (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methanol (620 mg, 1.41 mmol) and Et3N (0.39 mL, 2.8 mmol) in anhydrous DCM (12 mL) was added methanesulfonyl chloride (0.13 mL, 1.68 mmol) dropwise and the mixture was stirred at rt under N2 for 2 h. The reaction mixture was concentrated to dryness and the residue was purified by flash chromatography on silica gel (hexane / EtOAc) to give the title compound (684 mg) as a colorless oil. 1 H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.56(d,1H),7.38(dd,1H),4.33(s,2H),3.81-3.89(m,4H),3.73(s,2H),3.04(s,3H),0.85-0.90(m,9H),0.04-0.08(m,6H).

[0586] Step 6. 3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)-3-((3,4-difluorophenoxy)methyl)azetidine. A mixture of (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate (32.6 mg, 0.06 mmol), 3,4-difluorophenol (10.4 mg, 0.08 mmol) and K2CO3 (17.9 mg, 0.13 mmol) in anhydrous DMF (0.5 mL) was stirred at 85°C for 18 h. The reaction mixture was cooled, diluted with EtOAc (10 mL), washed with 2M K2CO3 (2×5 mL) and brine (2×5 mL), dried over Na2SO4, filtered and concentrated to dryness to give the crude title compound (31.8 mg).

[0587] Step 7. (1-((2,4-dichlorophenyl)sulfonyl)-3-((3,4-difluorophenoxy)methyl)azetidin-3-yl)methanol. To solid 3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)-3-((3,4-difluorophenoxy)methyl)azetidine (31.8 mg, 0.06 mmol) in anhydrous THF (0.5 mL) was added TBAF (1 M in THF, 0.175 mL) and the mixture was stirred at rt for 20 h. The reaction mixture was diluted with EtOAc (10 mL), washed with saturated aqueous NaHCO3 (2×5 mL) and brine (2×5 mL), dried over Na2SO4, filtered and concentrated to dryness. The residue was purified by flash chromatography on silica gel (hexanes / EtOAc) to give the title compound as a colorless oil (18.4 mg). 1 H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.57(d,1H),7.39(dd,1H),7.03-7.11(m,1H),6.66-6.72(m,1H),6 .54-6.59(m,1H),4.05(s,2H),3.91-3.97(m,4H),3.90(s,2H),1.65(br.s,1H). LCMS[M+H] + 438,440.

[0588] Example 88-Compound 88

[0589]

[0590] 3-Chloro-4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 3-chloro-4-hydroxybenzonitrile. 1 H NMR (500MHz, CDCl3) δppm7.98(d,1H),7.66(d,1H),7.53-7.59(m,2H),7.36-7.41(m,1H),7.00(d,1H),4.26(s,2H),3.92-4.03(m,6H),1.79(t,1H). LCMS[M+H] + 461,463.

[0591] Example 89-Compound 89

[0592]

[0593] 3-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)benzonitrile was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 3-cyanophenol. 1 H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.57(d,1H),7.36-7.42(m,2H),7.27-7.30(m,1H),7.08-7. 15(m,2H),4.13(s,2H),3.93-4.00(m,4H),3.91(s,2H),1.77(br.s,1H). LCMS[M+H] + 427,429.

[0594] Example 90-Compound 90

[0595]

[0596] Methyl 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)benzoate was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and methyl 4-hydroxybenzoate. 1 H NMR (500MHz, CDCl3) δppm7.97-8.01(m,3H),7.57(d,1H),7.39(dd,1H),6.85-6.90 (m,2H),4.16(s,2H),3.93-4.00(m,4H),3.92(s,2H),3.89(s,3H),1.72(br.s,1H). LCMS[M+H] + 460,462.

[0597] Example 91-Compound 91

[0598]

[0599] (1-((2,4-dichlorophenyl)sulfonyl)-3-((pyridin-4-yloxy)methyl)azetidin-3-yl)methanol was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 4-hydroxypyridine.1 H NMR (500MHz, CDCl3) δppm 8.37(d,2H),7.98(d,1H),7.56(d,1H),7.38(dd,1H),6.71(d,2H),4.12(s,2H),3.92-3.98(m,4H),3.90(s,2H). LCMS[M+H] + 403,405.

[0600] Example 92-Compound 92

[0601]

[0602] (1-((2,4-dichlorophenyl)sulfonyl)-3-((p-tolyloxy)methyl)azetidin-3-yl)methanol was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and p-cresol. 1 H NMR (500MHz, CDCl3) δppm7.98(d,1H),7.56(d,1H),7.38(dd,1H),7.08(d,2H), 6.74(d,2H),4.08(s,2H),3.91-3.96(m,4H),3.90(s,2H),2.29(s,3H),1.76(br s,1H). LCMS[M+H] + 416,418.

[0603] Example 93-Compound 93

[0604]

[0605] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)benzamide was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 4-hydroxybenzamide. 1 H NMR(500MHz,DMSO-d6)δppm 7.97(d,1H),7.94(d,1H),7.78-7.85(m,3H),7.67(dd,1H),7.14-7.18(br s,1H),6.79-6.84(m,2H),5.07-5.14(m,1H),4.00(s,2H),3.78-3.84(m,4H),3.53-3.56(m,2H). LCMS[M+H]+ 445,447.

[0606] Example 94-Compound 94

[0607]

[0608] (3-(((2-chloropyridin-4-yl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methanol was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 2-chloropyridin-4-ol. 1 H NMR(500MHz,CDCl3)δppm 8.20(d,1H),7.98(d,1H),7.57(d,1H),7.39(dd,1H),6.82(d,1H),6.72( dd,1H),4.16(s,2H),3.93-3.97(m,4H),3.91(d,2H),1.90-1.95(m,1H). LCMS[M+H] + 437,439.

[0609] Example 95-Compound 95

[0610]

[0611] (1-((2,4-dichlorophenyl)sulfonyl)-3-(((2-methylpyridin-4-yl)oxy)methyl)azetidin-3-yl)methanol was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 2-methylpyridin-4-ol. 1 HNMR(500MHz, CDCl3)δppm8.26(d,1H),7.98(d,1H),7.57(d,1H),7.39(dd,1H) ,6.56-6.64(m,2H),4.13(s,2H),3.91-3.97(m,4H),3.90(s,2H),2.52(s,3H). LCMS[M+H] + 417,419.

[0612] Example 96-Compound 96

[0613]

[0614] (1-((2,4-dichlorophenyl)sulfonyl)-3-((m-tolyloxy)methyl)azetidin-3-yl)methanol was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and m-cresol. 1 H NMR(500MHz, CDCl3)δppm7.98(d,1H),7.56(d,1H),7.38(dd,1H),7.17(t,1H),6.80(d,1H),6 .62-6.68(m,2H),4.08(s,2H),3.92-3.97(m,4H),3.91(s,2H),2.33(s,3H),1.83(br.s,1H). LCMS[M+H] + 416,418.

[0615] Example 97-Compound 97

[0616]

[0617] (1-((2,4-dichlorophenyl)sulfonyl)-3-((pyridin-3-yloxy)methyl)azetidin-3-yl)methanol was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 3-hydroxypyridine. 1 H NMR(500MHz,CDCl3)δppm 8.21-8.29(m,2H),7.98(d,1H),7.57(d,1H),7.39(dd,1H),7.21-7.24(m,1H),7. 15-7.18(m,1H),4.16(s,2H),3.94-3.99(m,4H),3.93(s,2H),1.94-2.37(m,1H). LCMS[M+H] + 403,405.

[0618] Example 98-Compound 98

[0619]

[0620] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-3-methoxybenzonitrile was prepared in a manner similar to that in Scheme 24 from (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate and 4-hydroxy-3-methoxybenzonitrile. 1 H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.55(d,1H),7.37(dd,1H),7.23-7.29(m,1H),7.08(d,1H),6.92(d,1H),4.23(s,2H),3.90-4.00(m,6H),3.84(s,3H).

[0621] Example 99-Compound 99

[0622]

[0623] (3-((4-cyano-3-ethoxyphenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl 4-bromobenzoate was prepared according to Reaction Scheme 25:

[0624]

[0625] Reagents: Step 1) 4M NaOH aqueous solution, THF, EtOH; 2) BH3·SMe2, THF; 3) 4-bromobenzoyl chloride, Et3N, DCM.

[0626] Step 1. 3-((4-cyano-3-ethoxyphenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid. To a solution of ethyl 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylate (1.96 g, 4.0 mmol) in THF (25 mL) and EtOH (25 mL) was added aqueous NaOH (4 M, 10 mL) and the mixture was stirred at rt under N2 for 16 h. The reaction mixture was concentrated in vacuo to remove most of the THF and EtOH, acidified with 1 M HCl (100 mL), diluted with water (100 mL), saturated with solid NaCl and extracted with DCM (3 x 150 mL). The extracts were dried over Na2SO4, filtered and concentrated in vacuo to give a mixture of 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid and the title compound (1.86 g).

[0627] Step 2. 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-ethoxybenzonitrile. To a mixture of 3-((4-cyano-3-fluorophenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid and 3-((4-cyano-3-ethoxyphenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidine-3-carboxylic acid (941 mg, 2.0 mmol) in anhydrous THF (20 mL) was added BH3·SMe2 solution (0.3 mL, 3.16 mmol) dropwise at 0°C under N2. The mixture was warmed to rt and stirred for 20 h. The reaction mixture is quenched with 4M NaOH (50mL), poured into saturated NaHCO3 aqueous solution (200mL) and extracted with EtOAc (2 × 350mL). The organic phase is washed with brine (250mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue is purified by flash chromatography (hexane / EtOAc), giving a mixture of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidine-3-yl)methoxy)-2-fluorobenzonitrile and the title compound (703mg) as a viscous colorless oil.

[0628] Step 3. (3-((4-cyano-3-ethoxyphenoxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl 4-bromobenzoate. To a solution of a mixture of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-ethoxybenzonitrile (1.94 g, 4.4 mmol) and EtN (0.95 mL, 6.82 mmol) in anhydrous DCM (45 mL) was added solid 4-bromobenzoyl chloride (1.15 g, 5.24 mmol) and the mixture was stirred at rt under N for 4 h. The reaction mixture was concentrated in vacuo and the residue was purified by flash chromatography (hexanes / EtOAc) to give the title compound as a white solid (103 mg). 1 H NMR(500MHz,CDCl3)ppm 7.98(d,1H),7.84-7.89(m,2H),7.57-7.63(m,2H),7.53(d,1H),7.48(d,1H),7.38(d d,1H),6.40-6.50(m,2H),4.56(s,2H),4.19(s,2H),4.03-4.14(m,6H),1.48(t,3H).

[0629] Example 100-Compound 100

[0630]

[0631] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidine-3-yl)methoxy)-2-ethoxybenzonitrile was prepared as described below. To a solution of 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidine-3-yl)methoxy)-2-ethoxybenzonitrile (74.6 mg, 0.11 mmol) in THF (0.5 mL) and EtOH (0.5 mL) was added an aqueous NaOH solution (1 M, 1.15 mL), and the mixture was stirred at rt for 1 h. The reaction mixture was diluted with EtOAc (20 mL) and washed with saturated NaHCO3 aqueous solution (3×15 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (hexane / EtOAc) to give the title compound (25.2 mg) as a white solid. 1 H NMR(500MHz,CDCl3)δppm 7.98(d,1H),7.56(d,1H),7.47(d,1H),7.38(dd,1H),6.47(dd,1H),6.43(d,1H),4.14(s ,2H),4.08-4.14(m,2H),3.91-4.00(m,4H),3.90(s,2H),1.66-1.72(m,1H),1.48(t,3H). LCMS[M+H] + 471,472.

[0632] Example 101-Compound 101

[0633]

[0634] 4-(((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methyl)amino)-2-fluorobenzonitrile was prepared according to Reaction Scheme 26:

[0635]

[0636] Reagents: Step 1) 4-amino-2-fluorobenzonitrile, LHMDS (1.0 M in THF), NaI, DMF, 70°C; 2) TBAF, THF.

[0637] Step 1. 4-(((3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)amino)-2-fluorobenzonitrile. To a mixture of 4-amino-2-fluorobenzonitrile (27.8 mg, 0.20 mmol) and NaI (5.6 mg, 0.037 mmol) in anhydrous DMF (0.4 mL) was added LHMDS solution (1.0 M in THF, 0.20 mL) at rt under N2. After stirring for 15 min, a solution of (3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl methanesulfonate (51 mg, 0.10 mmol) in anhydrous DMF (0.8 mL) was added dropwise and the mixture was stirred at 70 °C for 22 h. The reaction mixture was cooled, diluted with EtOAc (10 mL), washed with saturated aqueous NaHCO3 (10 mL), brine (2×10 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (EtOAc / hexanes) to give the title compound (4.3 mg) as a yellow oil. 1 H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.56(d,1H),7.38(dd,1H),7.32(dd,1H),6.32(dd,1H),6.28(dd,1H),5.19(t,1 H),3.82-3.85(m,2H),3.81(s,2H),3.77-3.80(m,2H),3.40(d,2H),0.92(s,9H),0.10(s,6H).

[0638] Step 2. 4-(((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methyl)amino)-2-fluorobenzonitrile. To a solution of 4-(((3-(((tert-butyldimethylsilyl)oxy)methyl)-1-((2,4-dichlorophenyl)sulfonyl)azetidin-3-yl)methyl)amino)-2-fluorobenzonitrile (4.3 mg, 0.01 mmol) in anhydrous THF (0.5 mL) was added TBAF solution (1.0 M in THF, 40 μL) dropwise and the mixture was stirred at rt for 5 h. The reaction mixture was dry loaded on silica gel and purified by flash chromatography (EtOAc / hexanes) to give the title compound (1.4 mg) as a yellow oil. 1H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.56(d,1H),7.38(dd,1H),7.33(t,1H),6.37(dd,1H),6.33(dd ,1H),5.11(s,1H),3.88-3.94(m,4H),3.82(d,2H),3.44(d,2H),3.35(s,1H). LCMS[M+H] + 443.9,446.1.

[0639] Example 102-Compound 102

[0640]

[0641] 4-((1-((2-bromo-4-methoxyphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-bromo-4-methoxybenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.00(d,1H),7.53(dd,1H),7.28(d,1H),6.92(dd,1H),6.76(dd,1H),6.70(dd,1H),4.16(s,2H),3.85-3.94(m,9H),1.68(br.s,1H).

[0642] Example 103-Compound 103

[0643]

[0644] 4-((1-((2-cyano-4-methylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-methylbenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.96(d,1H),7.70(s,1H),7.49-7.57(m,2H),6.75(dd,1H),6.66(dd,1H),4 .16(s,2H),3.92-4.01(m,4H),3.89(s,2H),2.50(s,3H),1.76(br.s.,1H).

[0645] Example 104-Compound 104

[0646]

[0647] 4-((1-((2-cyano-4-fluorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-fluorobenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm8.10(dd,1H),7.59(dd,1H),7.54(dd,1H),7.41-7.46(m,1H),6 .78(dd,1H),6.73(dd,1H),4.18(s,2H),3.97-4.04(m,4H),3.90(s,2H),1.75(br.s,1H).

[0648] Example 105-Compound 105

[0649]

[0650] 4-((1-((2-bromo-4-(trifluoromethoxy)phenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-bromo-4-trifluoromethoxybenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 8.13(d,1H),7.61(d,1H),7.53(dd,1H),7.26-7.32(m,1H),6.77(dd,1H),6 .73(dd,1H),4.18(s,2H),3.94-4.02(m,4H),3.92(s,2H),1.74(br.s.,1H).

[0651] Example 106-Compound 106

[0652]

[0653] 4-((1-((2-cyano-4-methoxyphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-methoxybenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.99(d,1H),7.52(dd,1H),7.35(d,1H),7.18(dd,1H),6.75(dd,1H),6.67(dd,1H),4.16(s,2H),3.90-3.99(m,7H),3.89(s,2H),1.82(br.s,1H).

[0654] Example 107-Compound 107

[0655]

[0656] 4-((1-((2-cyano-4-trifluoromethoxyphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-3-fluorophenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-trifluoromethoxybenzenesulfonyl chloride. 1 H NMR (500MHz, CDCl3) δppm8.14(d,1H),7.70(d,1H),7.50-7.59(m,2H),6.77(dd,1 H),6.73(dd,1H),4.19(s,2H),3.99-4.08(m,4H),3.91(s,2H),1.74(br.s.,1H).

[0657] Example 108-Compound 108

[0658]

[0659] 4-((1-((2,4-dichlorophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluoro-5-methoxybenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-5-fluoro-2-methoxyphenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2,4-dichlorobenzenesulfonyl chloride. 1H NMR(500MHz,CDCl3)δppm 7.97(d,1H),7.55(d,1H),7.37(dd,1H),6.95(d,1H),6.73(d,1H),4.21(s, 2H),3.96-4.00(m,2H),3.90-3.95(m,4H),3.82(s,3H),2.06-2.13(m,1H).

[0660] Example 109-Compound 109

[0661]

[0662] 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluoro-5-methoxybenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-5-fluoro-2-methoxyphenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyanobenzenesulfonyl chloride. 1 H NMR(500MHz,DMSO-d6)δppm 8.15(dd,1H),8.03(dd,1H),7.91-7.96(m,1H),7.86-7.91(m,1H),7.39(d,1H),7.19( d,1H),4.97(t,1H),4.08(s,2H),3.80(d,2H),3.75(s,3H),3.72(d,2H),3.40(d,2H).

[0663] Example 110-Compound 110

[0664]

[0665] 4-((1-((2-cyano-4-methylphenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluoro-5-methoxybenzonitrile was prepared in a manner similar to that in Scheme 23 from 3-((4-cyano-5-fluoro-2-methoxyphenoxy)methyl)-3-(hydroxymethyl)azetidine-1-cation trifluoroacetate and 2-cyano-4-methylbenzenesulfonyl chloride. 1 H NMR(500MHz,CDCl3)δppm 7.95(d,1H),7.68(s,1H),7.50-7.55(m,1H),6.94(d,1H),6.72(d,1H),4.22(s,2H),3.97 -4.01(m,2H),3.93-3.97(m,2H),3.90(s,2H),3.82(s,3H),2.48(s,3H),2.08(br.s.,1H).

[0666] Example 111-Compound 111

[0667]

[0668] 4-((1-((2-cyanophenyl)sulfonyl)-3-((oxetan-3-ylamino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and oxetan-3-amine. 1 H NMR(500MHz,CDCl3)δppm 8.09(dd,1H),7.90(dd,1H),7.74(m,2H),7.54(dd,1H),6.76(dd,1H),6.71(dd,1H),4. 80(t,2H),4.37(t,2H),4.16(s,2H),3.98-4.03(m,2H),3.88-3.96(m,3H),2.91(s,2H). LCMS[M+H] + 456.9.

[0669] Example 112-Compound 112

[0670]

[0671] 4-((1-((2-cyanophenyl)sulfonyl)-3-(((trans-4-hydroxycyclohexyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and trans-4-aminocyclohexanol. 1 H NMR(500MHz,CDCl3)δppm 8.08(dd,1H),7.90(dd,1H),7.69-7.79(m,2H),7.52(dd,1H),6.74(dd,1H),6.69(dd,1H),4.12(s,2H),3.94-3.99(m,2H ),3.91(d,2H),3.59(tt,1H),2.90-2.96(m,2H),2.38(tt,1H),1.83-1.99(m,4H),1.23-1.33(m,2H),0.99-1.10(m,2H).

[0672] Example 113-Compound 113

[0673]

[0674] 4-((1-((2-cyanophenyl)sulfonyl)-3-(((trans-3-hydroxycyclobutyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and trans-3-aminocyclobutan-1-ol. 1 HNMR(500MHz, CDCl3)δppm8.08(dd,1H),7.90(dd,1H),7.69-7.80(m,2H),7.53(dd,1H),6.75(dd,1H),6.69(dd,1H),4.39-4.46(m, 1H),4.11-4.15(m,2H),3.96-4.01(m,2H),3.88-3.93(m,2H),3.36-3.43(m,1H),2.81(s,2H),2.05-2.13(m,2H),1.96-2.04(m,2H). LCMS[M+H] + 470.9.

[0675] Example 114-Compound 114

[0676]

[0677] Ethyl 3-(((3-((4-cyano-3-fluorophenoxy)methyl)-1-((2-cyanophenyl)sulfonyl)azetidin-3-yl)methyl)amino)propanoate was prepared in a similar manner to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and ethyl 3-aminopropanoate. 1 H NMR(500MHz,CDCl3)δppm 8.08(dd,1H),7.90(dd,1H),7.69-7.79(m,2H),7.52(dd,1H),6.75(dd,1H),6.70(dd,1H),4.08-4.15( m,4H),3.95-4.00(m,2H),3.89-3.94(m,2H),2.93(s,2H),2.86(t,2H),2.44(t,2H),1.21-1.25(m,3H). LCMS[M+H] + 501.1.

[0678] Example 115-Compound 115

[0679]

[0680] 4-((1-((2-cyanophenyl)sulfonyl)-3-(((2,3-dihydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and 3-aminopropane-1,2-diol. 1 HNMR(500MHz, CDCl3)δppm8.05-8.11(m,1H),7.90(dd,1H),7.69-7.80(m,2H),7.49-7.55(m,1H),6.75(dd,1H),6.70(dd,1H),4.12 -4.16(m,2H),3.95-4.02(m,4H),3.77(m,1H),3.68-3.73(m,1H),3.56(dd,1H),2.92-3.03(m,2H),2.78-2.84(m,1H),2.73(dd,1H). LCMS[M+H] + 475.1.

[0681] Example 116-Compound 116

[0682]

[0683] 4-((1-((2-cyanophenyl)sulfonyl)-3-(((tetrahydro-2H-pyran-4-yl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and 4-aminotetrahydropyran. 1 H NMR(500MHz,CDCl3)δppm 8.09(dd,1H),7.90(dd,1H),7.69-7.79(m,2H),7.49-7.55(m,1H),6.75(dd,1H),6.69(dd,1H),4.12-4.16 (m,2H),3.89-4.00(m,8H),3.35(m,1H),2.96(s,2H),2.56-2.65(m,1H),1.73-1.82(m,2H),1.30(dd,2H). LCMS[M+H] + 484.5.

[0684] Example 117-Compound 117

[0685]

[0686] (S)-4-((1-((2-cyanophenyl)sulfonyl)-3-(((2-hydroxypropyl)amino)methyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile was prepared in a manner similar to that in Scheme 14 from 4-((1-((2-cyanophenyl)sulfonyl)-3-(hydroxymethyl)azetidin-3-yl)methoxy)-2-fluorobenzonitrile and (S)-1-aminopropan-2-ol. 1 H NMR(500MHz,CDCl3)δppm 8.08(dd,1H),7.90(dd,1H),7.69-7.79(m,2H),7.49-7.56(m,1H),6.76(dd,1H),6.70(dd,1H),4.13-4.17(m,2H ),3.94-4.02(m,4H),3.74-3.82(m,1H),2.91-3.05(m,2H),2.69-2.75(m,1H),2.46(dd,1H),1.12-1.16(m,3H). LCMS[M+H] + 458.8.

[0687] Example 118. Biological Activity

[0688] The biological activity of the TRPV4 antagonists of the present disclosure was evaluated as follows.

[0689] Evaluation of the activity and potency of TRPV4 antagonists in human embryonic kidney cells (IC 50 )

[0690] Perform cell-based Ca 2+ Fluorescence-based assay to assess TRPV4 antagonist activity and determine potency (IC) in human embryonic kidney 293 cells (HEK-293) stably overexpressing human TRPV4 50The assay was performed as described in Deering-Rice, CE, Johansen, ME, Roberts, JK, Thomas, KC, Romero, EG, Lee, J., Yost, GS, Veranth, JM and Reilly, CA (2012), Mol. Pharmacol. 81(3), 411-419; Shapiro, D., Deering-Rice, CE, Romero, EG, Hughen, RW, Light, AR, Veranth, JM and Reilly, CA (2013), Chem Res. Toxicol., 26, 750-758; and Deering-Rice, CE, Mitchell, VK, Romero, EG, Abdel Aziz, MH and Reilly, CA (2014), Pharmacol. Res. & Perspectives. 2(5), 1-10.

[0691] HEK-293 cells stably overexpressing human TRPV4 were grown to confluence in 2% (w / v) gelatin-coated 96-well plates in DMEM:F12 medium supplemented with 5% fetal bovine serum, 1× penicillin / streptomycin (Invitrogen), and geneticin (300 μg / mL).

[0692] HEK-293 cells were prepared for the assay by replacing the growth medium with a 1:1 solution of LHC-9 and Fluo4-Direct (Invitrogen) reagent containing Fluo 4-AM, pluronic F-127, probenecid and a proprietary quenching dye. The cells were then incubated at 37°C for 1 h in a cell culture incubator. The cells were then washed by replacing the loading solution with LHC-9 containing 1 mM water-soluble probenecid (Invitrogen), 750 μM trypan red (ATT Bioquest) and test compounds (various concentrations). The cells were incubated at 37°C for another 30 minutes to allow for the cleavage and activation of Fluo 4-AM and the equilibrium of both the test compounds and Fluo 4.

[0693] Agonist treatment solution (90 nM GSK1016790A, a potent activator of TRPV4) was prepared at 3× concentration in LHC9 and 25 μL was added to 50 μL of medium on cells in a 96-well plate to provide a final concentration of 30 nM. Changes in cell fluorescence were monitored for 1 min at 37°C using a BMGLabtech NOVOStar fluorescence plate reader. Data were quantified as the maximum rate of change in fluorescence intensity (maxΔF / s), corrected for medium-only treatment (negative control), and expressed as a percentage of response relative to the no antagonist control. Each treatment condition was repeated at least three times and at least 8 concentrations were used to derive the IC reported for each antagonist. 50 The IC values ​​of each antagonist are shown in Table 4 below. 50 Values ​​were calculated as described in Deering-Rice, CE, Johansen, ME, Roberts, JK, Thomas, KC, Romero, EG, Lee, J., Yost, GS, Veranth, JM and Reilly, CA (2012), Mol. Pharmacol. 81(3), 411-419; Shapiro, D., Deering-Rice, CE, Romero, EG, Hughen, RW, Light, AR, Veranth, JM and Reilly, CA (2013), Chem Res. Toxicol., 26, 750-758; and Deering-Rice, CE, Mitchell, VK, Romero, EG, Abdel Aziz, MH and Reilly, CA (2014), Pharmacol. Res. & Perspectives. 2(5), 1-10.

[0694] Table 4

[0695]

[0696]

[0697]

[0698]

[0699] TRPV4 selectivity

[0700] To determine the selectivity of various test compounds for TRPV4, the cell-based Ca 2+Inhibitory effects of selected compounds active in the flux TRPV4 fluorescence assay on human TRPA1 stably overexpressed in HEK-293 cells, TRPM8 stably overexpressed in HEK-293 cells, TRPV3 stably overexpressed in HEK-293 cells, and TRPV1 stably overexpressed in human lung epithelial (BEAS-2B) cells. The assay was performed essentially as described above for HEK-293 cells stably overexpressing TRPV4, although for TRPV1 BEAS-2B cells were maintained in LHC-9 medium containing geneticin (300 μg / mL) and loaded with Fluo-4AM at room temperature. The agonist for TRPA1 was 2,4-di-tert-butylphenol (50 μM), the agonist for TRPM8 was icilin (50 μM), the agonist for TRPV3 was carvacrol (300 μM), and the agonist for TRPV1 was vanillyl nonamide (5 μM). All other aspects of the above assay were identical. For compound 11, the IC values ​​for TRPA1, TRPM8, TRPV3 and TRPV1 were 50 The values ​​were 52.0 μM, 14.4 μM, >100 μM and 24.3 μM, respectively. For compound 52, the IC 50 The values ​​were 89.6 μM, 35.9 μM, >100 μM and 60.0 μM, respectively. These results show that compound 11 and compound 16 are significantly more selective for TRPV4. For compound 16, the IC values ​​for TRPA1, TRPM8, TRPV3 and TRPV1 are 50 The values ​​were 20.7 μM, 17.8 μM, >100 μM and 6.0 μM, respectively. For compound 17, the IC 50 The values ​​were >35.5 μM, 25.4 μM, >50 μM and 5.3 μM, respectively.

[0701] In vitro screening assay in ocular cells

[0702] Several test compounds were subjected to cell-based Ca2+ assays as described herein and in [Ryskamp et al., J Neurosci 34(47):15689, 2014; Ryskamp et al., Sci Rep 6:30583, 2016 and Jo et al., Proc Natl Acad Sci USA 113(14):3885, 2016]. 2+ Fluorescence flux measurements were performed to evaluate the TRPV4 antagonist activity and potency (IC) of the test compounds in primary human trabecular meshwork (TM) cells, Müller cells, and retinal microvascular endothelial cells. 50value).

[0703] The trabecular meshwork is a tissue region in the eye that is located around the base of the cornea, close to the ciliary body, and is responsible for draining aqueous humor from the eye through the anterior chamber. TM cells are believed to be the main source of resistance to the outflow of aqueous humor from the anterior eye and are therefore the main determinant of intraocular pressure (IOP), which is believed to be the main cause of glaucoma. An increase in IOP may increase the stiffness and contractility of TM cells by increasing the level of intracellular calcium. TRPV4 antagonists can reduce pressure-induced calcium elevations in TM cells, thereby reducing IOP by inhibiting their contractile responses. Primary TM cells were isolated from the juxtatubular and corneoscleral regions of the human eye and grown in trabecular meshwork cell culture medium (ScienCell, catalog number 6591) at 37°C and 5% CO2.

[0704] Müller cells are the main macroglial cells in the retina. Müller cells act as pressure and volume sensors, strongly express TRPV4 channels, and are believed to cause inflammatory responses associated with glaucomatous neurodegeneration and may cause glaucomatous neurodegeneration [Ryskamp et al., J Neurosci. 34 (47): 15689-700, 2014]. TRPV4 antagonists can reduce pressure-induced calcium-dependent reactive gliosis, glial swelling / edema, and inflammatory signaling in the retina. Müller glial cells and retinal ganglion cells (RGCs) were dissociated from mouse retinas digested in L-15 containing papain (7U / ml; Worthington) for 1 hour at room temperature.

[0705] Microvascular endothelial cells (MVEC) are key components of the blood-retinal barrier, regulating the entry of oxygen and metabolites into the inner retina and mediating functional hyperemia, light-induced retinal blood flow increase. The cells strongly express TRPV4 channels, which can transduce their responses to shear flow, swelling and stretching into changes in intracellular calcium concentration. TRPV4 antagonists appear to regulate retinal vascular tension and functional hyperemia responses. Primary MVEC isolated from human retinal capillary tissue was purchased from Cell Systems (ACBRI 181), and cells of passages 3-5 were grown in human endothelial growth medium (EBM-2, Lonza, CC-3156 & CC4176) at 37 ° C and 5% CO2.

[0706] Cell-based Ca 2+ was performed as described herein and in [Ryskamp et al., J Neurosci 34(47):15689, 2014, and Jo et al., J Neurosci 35(39):13525, 2015]. 2+Flux fluorescence measurements to assess TRPV4 antagonist activity in TM cells, Müller cells, and MVECs and determine potency (IC 50 Values). Activity after stimulation of cells with the TRPV4 selective agonist GSK1016790A (25 nM for TM cells and Müller cells, 10 nM for MVEC) was determined in the presence of various concentrations of different test compounds. More specifically, aliquots of 10 mM agonist compounds were prepared in DMSO and dissolved in saline buffer at test concentrations (10–25 nM). Calcium levels were determined in cells loaded with 5 μM Fura-2-AM (Life Technologies) calcium indicator dye for 15–30 min at room temperature and perfused with isotonic saline (pH 7.4) containing the following components (in mM): 98.5 NaCl, 5 KCl, 3 MgCl2, 2 CaCl2, 10 HEPES, 10 D-glucose, 93 mannitol. Calcium levels were imaged epifluorescently using an inverted Nikon Ti microscope with a 40x (1.3 NA oil and 0.8 NA water) objective and Nikon Elements software. Data were quantified as described in [Ryskamp et al., J Neurosci 34(47):15689, 2014, and Molnar et al., J Neurosci 36(11):3184]. The calculated IC values ​​shown in Table 4 below 50 Values ​​represent the mean of cells (3-6 slides) from at least three independent experiments using 20-100 cells per treatment condition.

[0707] The data showed that the calcium responses induced by the agonist (GSK1016790A) and swelling mediated by TRPV4 channels were inhibited by compound 11, with an IC 50 The values ​​were ∼0.01 μM to ∼0.03 μM and were inhibited by compound 52, IC 50 In contrast, HC067047 inhibited TRPV4-mediated calcium responses in ocular cells, with IC 50 Values ​​ranged from -0.5 to -1.6 μM. Differences in potency between different cell types may reflect different molecular conformations (heteromerization, phosphorylation) of TRPV4 channels.

[0708] In vivo assay to assess IOP reduction in a mouse model of ocular hypertension

[0709] Tonometry was used to measure IOP changes in a mouse model of ocular hypertension over the weeks following topical application of the compound.

[0710] To prepare the ocular hypertension mouse model, mice were anesthetized by intraperitoneal (IP) injection of ketamine / xylazine (90 mg / 10 mg / kg body weight). The eyes of the mice were anesthetized and dilated by applying 0.5% proparacaine hydrochloride eye drops and 1% tropicamide eye drops (Bausch & Lomb). The IOP in the ipsilateral eye was increased by injecting 2 μl of polystyrene microbeads (7.8 μm FluoSpheres; Bangs Laboratories) into the anterior chamber within 60 seconds using a Hamilton syringe. The contralateral eye was injected with phosphate buffered saline (PBS).

[0711] Following topical administration of compounds 11 and 52 and timolol, a known drug for the treatment of IOP associated with ocular hypertension and glaucoma, IOP was measured using rebound tonometer at 1, 2, 6, and 10 hours after injection. IOP was determined from the average of 10 to 20 readings according to the protocol described in Ryskamp et al., SciRep 6:30583, 2016. Figure 1 As shown in , topical administration of compounds 11 and 52 and 0.5% timolol both reduced IOP and protected RGCs in a mouse model of ocular hypertension. This confirms the effectiveness of the TRPV4 antagonist HC067047, which reduced IOP in the eyes of ocular hypertensive mice when applied at 0.3–1.0 mM [Ryskamp et al., Sci Rep 6:30583, 2016].

[0712] The in vivo data showed that a single topical application of low concentrations [1 μM and 10 μM] of compounds 11 and 52 effectively reduced IOP in the earliest measured time frame (1 hour). The effectiveness is comparable to that of 0.5% timolol, the standard IOP-lowering drug that targets the secondary (uveoscleral) outflow pathway in current clinical practice. TRPV4 blockers promote the drainage of aqueous humor through the conventional outflow pathway (Ryskamp et al., 2016) and can be used as an adjunct to timolol and / or latanoprost (Xalatan) to reduce IOP. Given that the administration of timolol is accompanied by significant side effects including blurred vision, droopy eyelids, eye burning, headache, drowsiness, numbness, dry mouth, nausea, diarrhea, mood changes, irregular heartbeat and loss of appetite, and that the prostaglandin analog (Xalatan) has been shown to cause blurred vision, eye burning / stinging / redness, headache, dizziness, joint pain, chest pain, browning of the iris, tearing, eyelash growth and vision changes when applied to lower IOP, the TRPV4 antagonists of the present disclosure may be used as alternatives to timolol, Xalatan and other drugs currently used to lower IOP.

[0713] In Vivo Assay to Assess IOP Lowering in a Dog Model of Ocular Hypertension

[0714] The present invention is further supported by the finding that compound 52 reduces IOP in ocular hypertensive dogs.

[0715] Beagles homozygous for an autosomal recessive missense mutation in the ADAMTS10 gene develop marked IOP elevation due to decreased aqueous humor outflow, resulting in open-angle glaucoma characterized by cupping of the optic nerve head and optic neuropathy. The carbonic anhydrase inhibitor dorzolamide hydrochloride produces an IOP reduction of 3.8-5.0 mm Hg in dogs with OAG (Gelatt KN, MacKay EO, Vet Ophthalmol. 2001 Mar;4(1):61-7), and the combination of dorzolamide and timolol achieves an IOP reduction of 6.6-8.4 mm Hg (Plummer CE, MacKay EO, Gelatt KN, Vet Ophthalmol. 2006 Jul-Aug;9(4):245-9; Scardillo A, Pugliese M, De Majo M, Niutta PP, Pugliese A., Vet Ther. 2010 11(3):E1-6), whereas IOP in glaucomatous dogs is not affected by Rho inhibitors (Leary KA, Lin KT, Steibel JP, Harman CD, Komáromy AM, Vet Ophthalmol. 24 December 2019).

[0716] Eight drops of Compound 52 (200 mM in hydroxypropylmethylcellulose solution, stock dissolved in DMSO) were administered to beagles at 7 AM for 1.5 hours. Control eyes received vehicle (hydroxypropylmethylcellulose eye drops appropriately diluted with DMSO). IOP was measured in dogs (N=5 OAG, N=5 asymptomatic dogs) under local anesthesia by tonometer at 9:00 AM (6 readings per event, average). General ophthalmic examinations were performed by slit lamp biomicroscopy (Kowa SL-17 portable slit lamp; Kowa Company, Ltd.) and indirect ophthalmoscopy (Keeler All Pupil II headset; Keeler Instruments; Pan Retinal 2.2D focusing lens; Volk Optical) to detect potential treatment-related adverse reactions.

[0717] Compound 52 was well tolerated by dogs (N=10) with no observable signs of ocular discomfort or conjunctival hyperemia. The mean baseline IOP was 45.5 mm Hg. Topical application of Compound 52 significantly lowered the IOP to a mean level of 36.6 mm Hg.

[0718] It should be understood that the above detailed description and the accompanying examples are merely illustrative and should not be taken as limiting the scope of the present invention, which is defined only by the claims and their equivalents.

[0719] Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications, including but not limited to changes and modifications related to the chemical structures, substituents, derivatives, intermediates, syntheses, compositions, formulations or methods of use of the invention, may be made without departing from the spirit and scope of the invention.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein: L 1 is -C(O)-, -S(O2)-, -S(O)- or C1-C4 alkylene; X is O or NR 1 ; m is 0, 1, 2, 3, or 4; n is 0, 1, 2, 3, or 4; Ar 1 and Ar 2 Each is independently selected from a 5-10 membered aryl or heteroaryl group; Z is CHR 2 R 3 , COR 4 or NHR 5 ; R 1 is selected from hydrogen and C1-C4 alkyl; R 2 , R 3 , R 4 and R 5 Each is independently selected from hydrogen, hydroxy, C1-C6 alkyl, hydroxy-C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C1-C6 alkyl, phosphate, C1-C6 alkyl phosphate, C4-C8 heterocyclyl, NR 6 R 7 、-OC(O)R 8 、-C(O)R 9 、-S(O)2R 10 and -OS(O)2R 11 ; R 6 , R 7 , R 8 , R 9 , R 10 and R 11 each independently selected from hydrogen, C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C4-C8 cycloalkyl, C4-C8 heterocyclyl, C5-C8 aryl, C5-C8 heteroaryl, C1-C6 alkoxy-C1-C6 alkyl, C5-C8 aryl-C1-C6 alkyl, C5-C8 heteroaryl-C1-C6 alkyl, C5-C8 heterocyclyl-C1-C6 alkyl, amino-C1-C6 alkyl, hydroxy-C1-C6 alkyl, hydroxy-C3-C8 cycloalkyl, cyano-C1-C6 alkyl, hydroxysulfonyl-C1-C6 alkyl, phosphate-C1-C6 alkyl, alkylphosphate-C1-C6 alkyl, -COR 12 and-CR 13 R 14 C(O)R 15 ; R 12 Selected from C1-C6 alkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, C4-C8 heterocyclyl, hydroxy-C1-C6 alkyl, amino-C4-C8 heterocyclyl, phosphate-C1-C6 alkyl and C1-C6 alkyl phosphate; R 13 and R 14 are each independently selected from hydrogen, C1-C6 alkyl and C1-C6 heteroalkyl, or optionally taken together with the atoms to which they are attached to form a ring; and R 15 is hydroxy, C1-C6 alkoxy or amino; wherein at least one of m and n is not zero, and each aryl, heteroaryl, cycloalkyl or heterocycle is independently unsubstituted or substituted with one or more substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy, amino-C1-C6 alkyl, haloalkyl, sulfonyl, -C(O)-C1-C6 alkoxy and -C(O)-NH2.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein L 1 It is -S(O2)-.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ar 1 is selected from a 5-membered aryl group, a 5-membered heteroaryl group, a 6-membered aryl group or a 6-membered heteroaryl group, wherein each aryl or heteroaryl is unsubstituted or substituted with one or two substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy, haloalkyl, sulfonyl and -C(O)-C1-C6 alkoxy.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ar 2 is selected from a 5-membered aryl group, a 5-membered heteroaryl group, a 6-membered aryl group or a 6-membered heteroaryl group, wherein each aryl or heteroaryl is unsubstituted or substituted with one or two substituents independently selected from cyano, halogen, C1-C6 alkyl, C1-C6 alkoxy, -C(O)-C1-C6 alkoxy and -C(O)-NH2.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Z is -CH2OH, -C(O)OH, NH2, -NHC(O)CH2, -CH2NH2, -CH2NHCH3, -CH2NH(CH2)2OH, -CH2NH(CH2)3OH, -CH2NH(C5H9O), -CH2NH(CH2)2OCH3, -CH2NHCH2(C4H7O), -CH2NHCH2CH3, -CH2NHCH2CH(OH)CH3, -CH2NH(C3H5O) or -CH2NHCH2(C3H5O).

6. The compound according to any one of claims 1 to 5, selected from:

7. A pharmaceutical composition comprising an effective amount of at least one compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

8. A method for treating a disorder associated with transient receptor potential cation channel subfamily V, member 4 ("TRPV4") receptor activity in a subject, the method comprising administering to the subject an effective amount of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 7.

9. The method of claim 8, wherein the disorder is ocular hypertension.

10. The method of claim 8, wherein the disorder is atherosclerosis, disorders associated with intestinal edema, postoperative abdominal edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone-related dysfunction and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator-induced lung injury, altitude-induced pulmonary edema, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, sinusitis / rhinitis, asthma, overactive bladder, pain, motor neuron disorders, inherited gain-of-function disorders, cardiovascular disease, renal insufficiency, osteoarthritis, Crohn's disease, colitis, diarrhea, bowel irregularities (hyper-responsiveness / hypo-responsiveness), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramping, celiac disease, lactose intolerance, or flatulence.

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