Compounds
11834433 · 2023-12-05
Assignee
Inventors
Cpc classification
C07D405/04
CHEMISTRY; METALLURGY
International classification
C07D401/12
CHEMISTRY; METALLURGY
C07D405/04
CHEMISTRY; METALLURGY
Abstract
The present invention relates to compounds of formula (I): comprising a 5-membered nitrogen-containing heteroaryl ring A, wherein the heteroaryl ring A is substituted with at least one group containing an oxygen atom, and wherein said oxygen atom is attached to the heteroaryl ring A via at least two other atoms. The present invention further relates to salts, solvates and prodrugs of such compounds, to pharmaceutical compositions comprising such compounds, and to the use of such compounds in the treatment and prevention of medical disorders and diseases, most especially by the inhibition of NLRP3. ##STR00001##
Claims
1. A compound of formula (I): ##STR00085## or a pharmaceutically acceptable salt or solvate thereof, wherein: ring A is monocyclic; Q is selected from O or S; V is selected from N or C, and W, X, Y and Z are each independently selected from N, O, NH or CH, wherein at least one of V, W, X, Y and Z is N or NH; L is a saturated or unsaturated hydrocarbylene group, wherein the hydrocarbylene group may be straight-chained or branched, or be or include cyclic groups, wherein the hydrocarbylene group may optionally be substituted, and wherein the hydrocarbylene group may optionally include one or more heteroatoms N, O or S in its carbon skeleton; R.sup.1 is hydrogen or a saturated or unsaturated hydrocarbyl group, wherein the hydrocarbyl group may be straight-chained or branched, or be or include cyclic groups, wherein the hydrocarbyl group may optionally be substituted, and wherein the hydrocarbyl group may optionally include one or more heteroatoms N, O or S in its carbon skeleton; optionally L and R.sup.1 together with the oxygen atom to which they are attached may form a 3- to 12-membered saturated or unsaturated cyclic group, wherein the cyclic group may optionally be substituted; R.sup.2 is a cyclic group substituted at the α and α′ positions, wherein R.sup.2 may optionally be further substituted; R.sup.3 is hydrogen, halogen, —OH, —NH.sub.2, —CN, —R.sup.5, —OR.sub.5, —NHR.sub.5 or —N(R.sup.5).sub.2; R.sup.4 is hydrogen, halogen, —OH, —NH.sub.2, —CN, —R.sup.5, —OR.sub.5, —NHR.sub.5 or —N(R.sup.5).sub.2; or R.sup.3 and R.sup.4 together with the carbon atom to which they are attached may form a 3- to 7-membered saturated or unsaturated cyclic group, wherein the cyclic group may optionally be substituted; each R.sup.5 is independently an optionally substituted C.sub.1-C.sub.4 alkyl group; each R.sup.6 is independently halogen, —OH, —NO.sub.2, —NH.sub.2, —N.sub.3, —SH, —SO.sub.2H, —SO.sub.2NH.sub.2, or a saturated or unsaturated hydrocarbyl group, wherein the hydrocarbyl group may be straight-chained or branched, or be or include cyclic groups, wherein the hydrocarbyl group may optionally be substituted, and wherein the hydrocarbyl group may optionally include one or more heteroatoms N, O or S in its carbon skeleton; and m is 0, 1, 2 or 3; provided that any atom of L or R.sup.1 that is directly attached to W, X, Y or Z is not the same atom of L or R.sup.1 that is directly attached to the oxygen atom of R.sup.1—O-L-.
2. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein W, X, Y and Z are each independently N, NH or CH.
3. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein at least two of V, W, X, Y and Z are N or NH.
4. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein the group R.sup.1—O-L-, including any optional substituents, contains only atoms selected from the group consisting of carbon, hydrogen, oxygen and halogen atoms.
5. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein the group ##STR00086## including any optional substituents, contains from 8 to 16 atoms other than hydrogen or halogen.
6. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein each R.sup.6 is independently selected from halo; —CN; —NO.sub.2; —N.sub.3; —R.sup.β; —OH; —OR.sup.β; —R.sup.α-halo; —R.sup.α—CN; —R.sup.α—NO.sub.2; —R.sup.α—N.sub.3; —R.sup.α—R.sup.β; —R.sup.α—OH; —R.sup.α—OR.sup.β; —SH; —SR.sup.β; —SOR.sup.β; —SO.sub.2H; —SO.sub.2R.sup.β; —SO.sub.2NH.sub.2; —SO.sub.2NHR.sup.β; —SO.sub.2N(R.sup.β).sub.2; —R.sup.α—SH; —R.sup.α—SR.sup.β; —R.sup.α—SOR.sup.β; —R.sup.α—SO.sub.2H; —R.sup.α—SO.sub.2R.sup.β; —R.sup.α—SO.sub.2NH.sub.2; —R.sup.α—SO.sub.2NHR.sup.β; —R.sup.α—SO.sub.2N(R.sup.β).sub.2; —NH.sub.2; —NHR.sup.β; —N(R.sup.β).sub.2; —R.sup.α—NH.sub.2; —R.sup.α—NHR.sup.β; —R.sup.α—N(R.sup.β).sub.2; —CHO; —COR.sup.β; —COOH; —COOR.sup.β; —OCOR.sup.β; —R.sup.α—CHO; —R.sup.α—COR.sup.β; —R.sup.α—COOH; —R.sup.α—COOR.sup.β; or —R.sup.α—OCOR.sup.β; wherein each —R.sup.α— is independently selected from an alkylene, alkenylene or alkynylene group, wherein the alkylene, alkenylene or alkynylene group contains from 1 to 6 atoms in its backbone, wherein one or more carbon atoms in the backbone of the alkylene, alkenylene or alkynylene group may optionally be replaced by one or more heteroatoms N, O or S, and wherein the alkylene, alkenylene or alkynylene group may optionally be substituted with one or more halo and/or —R.sup.β groups; and wherein each —R.sup.β is independently selected from a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.7 alkynyl or C.sub.3-C.sub.6 cyclic group, and wherein any —R.sup.β may optionally be substituted with one or more C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 haloalkyl, C.sub.3-C.sub.7 cycloalkyl, —O(C.sub.1-C.sub.4 alkyl), —O(C.sub.1-C.sub.4 haloalkyl), —O(C.sub.3-C.sub.7 cycloalkyl), halo, —OH, —NH.sub.2, —CN, —C≡CH, oxo (═O), or 4- to 6-membered heterocyclic group.
7. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein R.sup.2 is an aryl or a heteroaryl group, wherein the aryl or the heteroaryl group is substituted at the α and α′ positions, and wherein R.sup.2 may optionally be further substituted.
8. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 7, wherein R.sup.2 is a fused aryl or a fused heteroaryl group, wherein a first cycloalkyl, cycloalkenyl, non-aromatic heterocyclic, aryl or heteroaryl ring is fused to the aryl or heteroaryl group across the α,β positions and a second cycloalkyl, cycloalkenyl, non-aromatic heterocyclic, aryl or heteroaryl ring is fused to the aryl or heteroaryl group across the α′,β′ positions, and wherein R.sup.2 may optionally be further substituted.
9. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein R.sup.2 is a cyclic group substituted at the α-position with a monovalent heterocyclic group or a monovalent aromatic group, wherein a ring atom of the heterocyclic or aromatic group is directly attached to the α-ring atom of the cyclic group, wherein the heterocyclic or aromatic group may optionally be substituted, and wherein the cyclic group is substituted at the α′-position and may optionally be further substituted.
10. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein R.sup.3 and R.sup.4 are hydrogen.
11. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein Q is O.
12. The compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, wherein the compound is selected from the group consisting of: ##STR00087## ##STR00088## ##STR00089## ##STR00090## ##STR00091## ##STR00092## ##STR00093## ##STR00094## ##STR00095## ##STR00096## and pharmaceutically acceptable salts and solvates thereof.
13. A pharmaceutical composition comprising the compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, and a pharmaceutically acceptable excipient.
14. A method of treating, delaying onset of, or reducing risk of a disease, disorder or condition in a subject, the method comprising the step of administering an effective amount of the compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1 to the subject, thereby treating, delaying onset of, or reducing risk of the disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.
15. The method as claimed in claim 14, wherein the disease, disorder or condition is selected from: (i) inflammation; (ii) an auto-immune disease; (iii) cancer; (iv) an infection; (v) a central nervous system disease; (vi) a metabolic disease; (vii) a cardiovascular disease; (viii) a respiratory disease; (ix) a liver disease; (x) a renal disease; (xi) an ocular disease; (xii) a skin disease; (xiii) a lymphatic condition; (xiv) a psychological disorder; (xv) graft versus host disease; (xvi) allodynia; and (xvii) any disease where an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.
16. The method as claimed in claim 14, wherein the disease, disorder or condition is selected from: (i) cryopyrin-associated periodic syndromes (CAPS); (ii) Muckle-Wells syndrome (MWS); (iii) familial cold autoinflammatory syndrome (FCAS); (iv) neonatal onset multisystem inflammatory disease (NOMID); (v) familial Mediterranean fever (FMF); (vi) pyogenic arthritis, pyoderma gangrenosum and acne syndrome (PAPA); (vii) hyperimmunoglobulinemia D and periodic fever syndrome (HIDS); (viii) Tumour Necrosis Factor (TNF) Receptor-Associated Periodic Syndrome (TRAPS); (ix) systemic juvenile idiopathic arthritis; (x) adult-onset Still's disease (AOSD); (xi) relapsing polychondritis; (xii) Schnitzler's syndrome; (xiii) Sweet's syndrome; (xiv) Behcet's disease; (xv) anti-synthetase syndrome; (xvi) deficiency of interleukin 1 receptor antagonist (DIRA); and (xvii) haploinsufficiency of A20 (HA20).
17. The method as claimed in claim 14, wherein the compound is administered as a pharmaceutical composition further comprising a pharmaceutically acceptable excipient.
18. A method of inhibiting NLRP3 in a subject, comprising administering the compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1 to the subject thereby inhibiting NLRP3.
19. A method of analysing inhibition of NLRP3 or an effect of inhibition of NLRP3 by a compound, comprising contacting a cell or non-human animal with the compound or the pharmaceutically acceptable salt or solvate thereof as claimed in claim 1, and analysing inhibition of NLRP3 or an effect of inhibition of NLRP3 in the cell or non-human animal by the compound.
Description
EXAMPLES—COMPOUND SYNTHESIS
(1) All solvents, reagents and compounds were purchased and used without further purification unless stated otherwise.
Abbreviations
(2) 2-MeTHF 2-methyltetrahydrofuran Ac.sub.2O acetic anhydride AcOH acetic acid aq aqueous Boc tert-butyloxycarbonyl br broad Cbz carboxybenzyl CDI 1,1-carbonyl-diimidazole conc concentrated d doublet DABCO 1,4-diazabicyclo[2.2.2]octane DAST diethylaminosulfur trifluoride DCE 1,2-dichloroethane, also called ethylene dichloride DCM dichloromethane DIPEA N,N-diisopropylethylamine, also called Hünig's base DMA dimethylacetamide DMAP 4-dimethylaminopyridine, also called N,N-dimethylpyridin-4-amine DME dimethoxyethane DMF N,N-dimethylformamide DMSO dimethyl sulfoxide EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide eq or equiv equivalent (ES.sup.+) electrospray ionization, positive mode Et ethyl EtOAc ethyl acetate EtOH ethanol h hour(s) HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate HPLC high performance liquid chromatography LC liquid chromatography m multiplet m-CPBA 3-chloroperoxybenzoic acid Me methyl MeCN acetonitrile MeOH methanol (M+H).sup.+ protonated molecular ion MHz megahertz min minute(s) MS mass spectrometry Ms mesyl, also called methanesulfonyl MsCl mesyl chloride, also called methanesulfonyl chloride MTBE methyl tert-butyl ether, also called tert-butyl methyl ether m/z mass-to-charge ratio NaHMDS sodium hexamethyldisilazide, also called sodium bis(trimethylsilyl)amide NaO.sup.tBu sodium tert-butoxide NBS 1-bromopyrrolidine-2,5-dione, also called N-bromosuccinimide NCS 1-chloropyrrolidine-2,5-dione, also called N-chlorosuccinimide NMP N-methylpyrrolidine NMR nuclear magnetic resonance (spectroscopy) Pd(OAc).sub.2 palladium acetate Pd(dba).sub.2 bis(dibenzylideneacetone) palladium(0) Pd.sub.2(dba).sub.3 tris(dibenzylideneacetone) dipalladium(0) Pd(dppf)Cl.sub.2 [1,1′-bis(diphenylphosphino)ferrocene] dichloropalladium(II) PE petroleum ether Ph phenyl PMB p-methoxybenzyl, also called 4-methoxybenzyl prep-HPLC preparative high performance liquid chromatography prep-TLC preparative thin layer chromatography PTSA p-toluenesulfonic acid q quartet RP reversed phase RT room temperature s singlet sat saturated SCX solid supported cation exchange (resin) SEM 2-(trimethylsilyl)ethoxymethyl sept septuplet t triplet T3P propylphosphonicanhydride TBME tert-butyl methyl ether, also called methyl tert-butyl ether TBSCl tert-butyldimethylsilyl chloride TEA triethylamine TFAA 2,2,2-trifluoroacetic acid anhydride TFA 2,2,2-trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography TMSCl trimethylsilyl chloride wt % weight percent or percent by weight XantPhos® 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene Xphos® 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl XtalFluor-E® (diethylamino)difluorosulfonium tetrafluoroborate
(3) Experimental Methods
(4) Analytical Methods
(5) NMR spectra were recorded at 300, 400 or 500 MHz with chemical shifts reported in parts per million. Spectra were measured at 298 K, unless indicated otherwise, and were referenced relative to the solvent resonance. Spectra were collected using one of the machines below:— An Agilent VNMRS 300 instrument fitted with a 7.05 Tesla magnet from Oxford instruments, indirect detection probe and direct drive console including PFG module. An Agilent MercuryPlus 300 instrument fitted with a 7.05 Tesla magnet from Oxford instruments, 4 nuclei auto-switchable probe and Mercury plus console. A Bruker Avance III spectrometer at 400 MHz fitted with a BBO 5 mm liquid probe. A Bruker 400 MHz spectrometer using ICON-NMR, under TopSpin program control. A Bruker Avance III HD spectrometer at 500 MHz, equipped with a Bruker 5 mm SmartProbe™.
(6) HPLC and LC-MS were recorded on an Agilent 1290 series with UV detector and HP 6130 MSD mass detector. Mobile phase A: ammonium acetate (10 mM); water/MeOH/acetonitrile (900:60:40); mobile phase B: ammonium acetate (10 mM); water/MeOH/acetonitrile (100:540:360); column, Waters XBridge BEH C18 XP (2.1×50 mm, 2.5 μm).
(7) TABLE-US-00001 Pump flow: 0.6 mL/min UV detection: 215, 238 nm Injection volume: 0.2 μL Run time: 4.0 min Column temperature: 35° C. Mass detection: API-ES +ve and −ive
(8) Pump Program:
(9) TABLE-US-00002 Gradient Time (min) % A % B 0.0 80 20 0.5 80 20 2.0 0 100
(10) Alternatively LC-MS were recorded using SHIMADZU LCMS-2020, Agilent 1200 LC/G1956A MSD and Agilent 1200\G6110A, or Agilent 1200 LC & Agilent 6110 MSD. Mobile Phase: A: 0.025% NH.sub.3.Math.H.sub.2O in water (v/v); B: Acetonitrile. Column: Kinetex EVO C18 2.1×30 mm, 5 μm.
(11) Reversed Phase HPLC Conditions for the LCMS Analytical Methods
(12) Methods 1a and 1b: Waters Xselect CSH C18 XP column (4.6×30 mm, 2.5 μm) at 40° C.; flow rate 2.5-4.5 mL min-1 eluted with a H.sub.2O-MeCN gradient containing either 0.1% v/v formic acid (Method 1a) or 10 mM NH.sub.4HCO.sub.3 in water (Method 1b) over 4 min employing UV detection at 254 nm.
(13) Method 1c: Agilent 1290 series with UV detector and HP 6130 MSD mass detector using Waters XBridge BEH C18 XP column (2.1×50 mm, 2.5 μm) at 35° C.; flow rate 0.6 mL/min; mobile phase A: ammonium acetate (10 mM); water/MeOH/acetonitrile (900:60:40); mobile phase B: ammonium acetate (10 mM); water/MeOH/acetonitrile (100:540:360); over 4 min employing UV detection at 215 and 238 nm.
(14) Reversed Phase HPLC Conditions for the UPLC Analytical Methods
(15) Methods 2a and 2b: Waters BEH C18 (2.1×30 mm, 1.7 μm) at 40° C.; flow rate 0.77 mL min.sup.−1 eluted with a H.sub.2O-MeCN gradient containing either 0.1% v/v formic acid (Method 2a) or 10 mM NH.sub.4HCO.sub.3 in water (Method 2b) over 3 min employing UV detection at 254 nm.
(16) Purification Method 1
(17) Automated reversed phase column chromatography was carried out using a Buchi Sepracore® X50 system driven by a C-605 pump module, C-620 Sepracore control package, C-640 UV photometer detection unit and C-660 fraction collector.
(18) Revelis C18 Reversed-Phase 12 g Cartridge
(19) TABLE-US-00003 Carbon loading 18% Surface area 568 m.sup.2/g Pore diameter 65 Angstrom pH (5% slurry) 5.1 Average particle size 40 μm
(20) The column was conditioned before use with MeOH (5 min), then brought to H.sub.2O (in 5 min) and kept 5 min at H.sub.2O. Flow rate=30 mL/min.
(21) Separation runs:
(22) TABLE-US-00004 Time (min) A: water (%) B: MeOH (%) 0 100 0 5 100 0 30 30 70 30.1 0 100 35 0 100
(23) Detection wavelength: 215, 235, 254 and 280 nm. Before each new run, the cartridge was cleaned using the conditioning method.
(24) Purification Method 2
(25) Preparative column chromatography was carried out using a Waters prep system driven by a 2767 Sample Manager, SFO System Fluidics Organizer, 515 HPLC Pumps, 2545 Binary Gradient Module, 2998 Photodiode Array Detector, SQD Detector 2 with ESI mass. Mobile phase ACD: acetonitrile; mobile phase A: ammonium acetate (10 mM); mobile phase B: acetonitrile; column, XSelect CSH Prep C18 OBD (100×30 mm; 5 μm).
(26) TABLE-US-00005 Pump flow: 40 mL/min Injection volume: 1.5 mL Run time: 15.0 min Column temperature: not controlled Mass detection: API-ES +ve and −ive
(27) Pump Program:
(28) TABLE-US-00006 Flow (ml/min) Flow (ml/min) Time (min) Bin. pump ACD pump % A % B 0.0 22 4 85 15 2.0 38 2 85 15 2.5 38 2 85 15 10.0 38 2 65 35 10.1 38 2 5 95 12.0 38 2 5 95 12.1 38 2 85 15 15.0 38 2 85 15
(29) Purification Method 3 (Acidic Prep)
(30) Preparative reversed phase HPLC was carried out using a Waters X-Select CSH column C18, 5 μm (19×50 mm), flow rate 28 mL min-1 eluting with a H.sub.2O-MeCN gradient containing 0.1% v/v formic acid over 6.5 min using UV detection at 254 nm. Gradient information: 0.0-0.2 min, 20% MeCN; 0.2-5.5 min, ramped from 20% MeCN to 40% MeCN; 5.5-5.6 min, ramped from 40% MeCN to 95% MeCN; 5.6-6.5 min, held at 95% MeCN.
(31) Purification Method 4 (Basic Prep)
(32) Preparative reversed phase HPLC was carried out using a Waters X-Bridge Prep column C18, 5 μm (19×50 mm), flow rate 28 mL min-1 eluting with a 10 mM NH.sub.4HCO.sub.3-MeCN gradient over 6.5 min using UV detection at 254 nm. Gradient information: 0.0-0.2 min, 10% MeCN; 0.2-5.5 min, ramped from 10% MeCN to 40% MeCN; 5.5-5.6 min, ramped from 40% MeCN to 95% MeCN; 5.6-6.5 min, held at 95% MeCN.
(33) Alternatively automated reversed phase HPLC column chromatography purification was carried out using: (i) a Gilson GX-281 system driven by a Gilson-322 pump module, Gilson-156 UV photometer detection unit and Gilson-281 fraction collector. Detection wavelength: 220 nm and 254 nm and 215 nm. (ii) a Gilson GX-215 system driven by a LC-20AP pump module, SPD-20A UV photometer detection unit and Gilson-215 fraction collector. Detection wavelength: 220 nm and 254 nm and 215 nm. (iii) a TELEDYNE ISCO CombiFlash Rf+150. Detection wavelength: 220 nm and 254 nm and 215 nm. (iv) a Shimadzu CBM-20A system driven by LC-20AP pump module, SPD-20A UV photometer detection unit and FRC-10A fraction collector. Detection wavelength: 220 nm and 254 nm and 215 nm.
Synthesis of Intermediates
Intermediate A1: 2-(5-(2-Cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetyl chloride
Step A: 5-(Benzyloxy)-4-bromo-2,3-dihydro-1H-indene
(34) ##STR00047##
(35) To a solution of 4-bromo-2,3-dihydro-1H-inden-5-ol (1.36 g, 6.38 mmol, 1 eq) (Hunsberger et al., JACS, 1955, vol. 77(9), pages 2466-2475) in dimethylformamide (35 mL) was added potassium carbonate (1.76 g, 12.8 mmol, 2 eq) and benzyl bromide (0.83 mL, 7.02 mmol, 1.1 eq). The reaction mixture was heated to 60° C. After stirring for 1.5 hours, the mixture was cooled to room temperature and diluted with diethyl ether. The organic layer was washed 4 times with water, once with brine, dried over sodium sulfate and then concentrated in vacuo to afford the title compound (1.83 g, 6.04 mmol, 94%).
(36) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.52-7.46 (m, 2H), 7.42-7.29 (m, 3H), 7.03 (d, 1H), 6.72 (d, 1H), 5.13 (s, 2H), 2.96 (t, 4H), 2.10 (p, 2H).
Step B: tert-Butyl 2-(5-(benzyloxy)-2,3-dihydro-1H-inden-4-yl)acetate
(37) ##STR00048##
(38) A solution of 5-(benzyloxy)-4-bromo-2,3-dihydro-1H-indene (1.83 g, 6.04 mmol, 1 eq) in anhydrous tetrahydrofuran (50 mL) was bubbled through with nitrogen for 20 minutes. To the degassed solution was added tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct (312 mg, 302 μmol, 0.05 eq) and dicyclohexyl(2′,4′,6′-triisopropyl-[1,1′-biphenyl]-2-yl)phosphane (288 mg, 604 μmol, 0.1 eq). The reaction mixture was stirred for 30 minutes at room temperature. After that, (2-(tert-butoxy)-2-oxoethyl) zinc (II) bromide (Intermediate A2, Step D) in THF (0.55 molar, 22 mL, 12.1 mmol, 2 eq) was added and the reaction mixture was heated in a sand bath at 70° C. After stirring for 1 hour, the reaction mixture was cooled to room temperature and then diluted with diethyl ether. The reaction mixture was washed twice with saturated ammonium chloride, once with brine, dried over sodium sulfate, filtered and then concentrated in vacuo. The crude product was submitted to normal phase flash chromatography using heptane and ethyl acetate as eluent to afford the title compound (1.82 g, 5.38 mmol, 89%).
(39) .sup.1H NMR (300 MHz, CDCl.sub.3) 7.44 (d, 2H), 7.40-7.29 (m 3H), 7.05 (d, 1H), 6.72 (d, 1H), 5.06 (s, 2H), 3.62 (s, 2H), 2.87 (t, 4H), 2.08 (p, 2H), 1.40 (s, 9H).
Step C: tert-Butyl 2-(5-hydroxy-2,3-dihydro-1H-inden-4-yl)acetate
(40) ##STR00049##
(41) A solution of tert-butyl 2-(5-(benzyloxy)-2,3-dihydro-1H-inden-4-yl)acetate (1.82 g, 5.38 mmol, 1 eq) in 2,2,2-trifluoroethanol (50 mL) was bubbled through with nitrogen for 20 minutes. After that, Pd/C (10 wt % loading, matrix activated carbon support, 0.57 g, 538 μmol, 0.1 eq) was added and the flask was charged with hydrogen. The reaction mixture was stirred under a hydrogen atmosphere. After 1.5 hours of stirring, another batch of Pd/C (10 wt % loading, matrix activated carbon support, 0.57 g, 538 μmol, 0.1 eq) was added. After stirring over the weekend, the reaction mixture was filtered over Celite®, and the residue was washed extensively with ethyl acetate. The filtrates were combined and concentrated in vacuo to afford the title compound (1.28 g, 5.15 mmol, 95%).
(42) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.33 (bs, 1H), 7.01 (d, 1H), 6.76 (d, 1H), 3.57 (s, 2H), 2.88 (td, 4H), 2.15-1.96 (m, 2H), 1.46 (s, 9H).
Step D: tert-Butyl 2-(5-(((trifluoromethyl)sulfonyl)oxy)-2,3-dihydro-1H-inden-4-yl)acetate
(43) ##STR00050##
(44) A solution of tert-butyl 2-(5-hydroxy-2,3-dihydro-1H-inden-4-yl)acetate (1.28 g, 5.15 mmol, 1 eq) and triethylamine (1.4 mL, 10.3 mmol, 2 eq) in dichloromethane (50 mL) was cooled in an ice bath. To the cooled greenish solution was added dropwise triflic anhydride (0.87 mL, 5.15 mmol, 1 eq). After complete addition, the cooling bath was removed and the reaction mixture was allowed to reach room temperature. After 1 hour of stirring, the reaction mixture was washed three times with saturated sodium bicarbonate solution, once with brine, dried over sodium sulfate, filtered and then concentrated in vacuo to afford the title compound (1.74 g, 4.57 mmol, 88%).
(45) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.17 (d, 1H), 7.07 (d, 1H), 3.63 (s, 2H), 2.92 (dt, 4H), 2.14 (p, 2H), 1.44 (s, 9H).
Step E: tert-Butyl 2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-4-yl)acetate
(46) ##STR00051##
(47) A solution of tert-butyl 2-(5-(((trifluoromethyl)sulfonyl)oxy)-2,3-dihydro-1H-inden-4-yl)acetate (4.64 g, 12.2 mmol, 1 eq) in 1,4-dioxane (61 mL) was degassed with nitrogen. After that, 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.3 mL, 36.6 mmol, 3.9 eq), triethylamine (10 mL, 73.2 mmol, 6.0 eq) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) adduct (498 mg, 610 μmol, 0.05 eq) was added. The reaction mixture was heated in a sand bath set at 100° C. After stirring overnight, the reaction mixture was concentrated in vacuo. The crude product was submitted to normal phase flash chromatography using heptane and ethyl acetate as eluent to afford the title compound (3.79 g, 10.5 mmol, 86%).
(48) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.63 (d, 1H), 7.13 (d, 1H), 3.92 (d, 2H), 2.90 (dt, 4H), 2.04 (p, 2H), 1.42 (s, 9H), 1.32 (s, 12H).
Step F: tert-Butyl 2-(5-(2-cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetate
(49) ##STR00052##
(50) A solution of tert-butyl 2-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-4-yl)acetate (3.74 g, 10.4 mmol, 1 eq) and 4-bromopicolinonitrile (2.29 g, 12.5 mmol, 1.2 eq) in acetonitrile (74 mL) and water (30 mL) was degassed with nitrogen. Then sodium carbonate (1.77 g, 16.7 mmol, 1.6 eq) and [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) adduct (852 mg, 1.04 mmol, 0.1 eq) were added. The reaction mixture was heated in a sand bath set at 80° C. After 50 minutes, the reaction mixture was cooled to room temperature, diluted with water and then extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, filtered and then concentrated in vacuo. The crude product was submitted to normal phase flash chromatography using ethyl acetate and heptane as eluent to afford the title compound (2.63 g, 7.86 mmol, 75%).
(51) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 8.71 (dd, 1H), 7.72 (dd, 1H), 7.52 (dd, 1H), 7.24 (d, 1H), 7.01 (d, 1H), 3.42 (s, 2H), 3.01 (t, 2H), 2.92 (t, 2H), 2.15 (p, 2H), 1.43 (s, 9H).
Step G: 2-(5-(2-Cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid TFA Salt and 2-(5-(2-carbamoylpyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid TFA Salt in Ratio ˜7:3
(52) ##STR00053##
(53) To a solution of tert-butyl 2-(5-(2-cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetate (2.63 g, 7.86 mmol, 1 eq) in dichloromethane (20 mL) was added trifluoroacetic acid (20 mL, 0.26 mol, 33 eq). The reaction mixture was stirred at room temperature for 2.5 hours and then toluene (40 mL) was added. The reaction mixture was concentrated to about 40 mL, and then again toluene (40 mL) was added; this process was done twice. Then all solvents were evaporated in vacuo to afford 2-(5-(2-cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid (2.92 g, 94%) as a ˜7:3 mixture with 2-(5-(2-carbamoylpyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid both as the TFA salt.
(54) .sup.1H NMR (of 2-(5-(2-cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid) (300 MHz, CDCl.sub.3) δ 8.78 (d, 1H), 7.74 (d, 1H), 7.58 (dd, 1H), 7.34-7.25 (m, 1H), 7.03 (d, 1H), 3.58 (d, 2H), 3.04 (d, 2H), 2.94 (t, 2H), 2.17 (p, 2H).
Step H: 2-(5-(2-Cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetyl chloride
(55) ##STR00054##
(56) To a solution of 2-(5-(2-cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid (non salt form) (34 mg, 0.12 mmol, 1 eq) in anhydrous dichloromethane (2 mL) was added one drop of dimethylformamide and after that dropwise oxalyl chloride (32 μL, 0.37 mmol, 3 eq) at room temperature. After stirring for 1 hour, the volatiles were removed in vacuo and the crude product was used for the following step without any purification.
Intermediate A2: 2-(4-Fluoro-2,6-diisopropylphenyl)acetic acid
Step A: 4-Fluoro-2,6-di(prop-1-en-2-yl)aniline
(57) ##STR00055##
(58) A solution of 2,6-dibromo-4-fluoroaniline (10 g, 1 eq), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (16.67 g, 2.67 eq), Cs.sub.2CO.sub.3 (36.35 g, 3 eq) and Pd(dppf)Cl.sub.2 (2.72 g, 3.72 mmol, 0.1 eq) in dioxane (100 mL) and H.sub.2O (10 mL) was degassed under reduced pressure. The reaction mixture was heated to 100° C. for 3 hours under nitrogen. Then the reaction mixture was quenched by addition of H.sub.2O (200 mL), diluted with EtOAc (150 mL), and extracted with EtOAc (2×150 mL). The combined organic layers were washed with brine (2×200 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 1:0 to 100:1) to give the title compound (8 g, 89% yield, 78.9% purity on LCMS) as a yellow oil.
(59) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.68 (d, 2H), 5.32-5.31 (m, 2H), 5.08 (d, 2H), 3.84 (s, 2H) and 2.07 (d, 6H).
(60) LCMS: m/z 192.2 (M+H).sup.+ (ES.sup.+).
Step B: 4-Fluoro-2,6-diisopropylaniline
(61) ##STR00056##
(62) To a solution of 4-fluoro-2,6-di(prop-1-en-2-yl)aniline (8 g, 1 eq) in MeOH (150 mL) was added Pd/C (624 mg, 10 wt % loading on activated carbon). The reaction mixture was degassed and purged with H.sub.2 (20 psi). The reaction mixture was stirred at 25° C. for 12 hours under H.sub.2 (20 psi), and then filtered. The filtrate was concentrated in vacuo. The residue was purified by column chromatography (SiO.sub.2, only eluting with petroleum ether) to give the title compound (4 g, 63% yield, 100% purity on LCMS) as a colourless oil.
(63) 1H NMR (400 MHz, CDCl.sub.3) δ 6.76 (d, 2H), 3.56 (s, 2H), 2.99-2.89 (m, 2H) and 1.26 (d, 12H).
(64) LCMS: m/z 196.2 (M+H).sup.+ (ES.sup.+).
Step C: 2-Bromo-5-fluoro-1,3-diisopropylbenzene
(65) ##STR00057##
(66) To a solution of 4-fluoro-2,6-diisopropylaniline (3.7 g, 18.95 mmol, 1 eq) in MeCN (180 mL) was added CuBr (4.08 g, 1.5 eq). Then tert-butyl nitrite (2.93 g, 1.5 eq) was added dropwise at 0° C. The resulting mixture was stirred at 60° C. for 1.5 hours, and then concentrated in vacuo. The residue was purified by column chromatography (SiO.sub.2, only eluting with petroleum ether) to give the title compound (2.02 g, 41%) as a white solid.
(67) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.85 (d, 2H), 3.55-3.48 (m, 2H) and 1.24 (d, 12H).
Step D: (2-(tert-Butoxy)-2-oxoethyl) zinc (II) bromide
(68) ##STR00058##
(69) A mixture of zinc (80 g) in HCl (1 M, 308 mL) was stirred at 25° C. for 30 minutes. Then the mixture was filtered and the filter cake was dried in vacuo. To a mixture of the above Zn (55 g, 841.11 mmol, 2.98 eq) in THF (550 mL) was added TMSCl (3.06 g, 28.20 mmol, 0.1 eq) and 1,2-dibromoethane (5.30 g, 28.20 mmol, 0.1 eq) at 20° C. under N.sub.2 atmosphere. Then tert-butyl 2-bromoacetate (55 g, 281.97 mmol, 1 eq) was added at 50° C. under N.sub.2 atmosphere. The reaction mixture was stirred at 50° C. for 2 hours. Then the reaction mixture (theory amount: 0.5 M, 550 mL, in THF solution) was cooled and used into the next step without further purification.
Step E: tert-Butyl 2-(4-fluoro-2,6-diisopropylphenyl)acetate
(70) ##STR00059##
(71) A solution of 2-bromo-5-fluoro-1,3-diisopropylbenzene (16 g, 61.74 mmol, 1 eq) in THF (100 mL) was cooled to 0° C. Then Pd.sub.2(dba).sub.3 (2.83 g, 3.09 mmol, 0.05 eq), Xphos (2.94 g, 6.17 mmol, 0.1 eq) and (2-(tert-butoxy)-2-oxoethyl) zinc (II) bromide (0.5 M, 246.95 mL, in THF solution, 2 eq) were added. The reaction mixture was stirred at 70° C. for 12 hours, and then concentrated in vacuo. The residue was purified by column chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 100:0 to 10:1) to give the title compound (12 g, 59% yield, 90% purity on .sup.1H NMR) as a red oil.
(72) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 6.83 (d, 2H), 3.66 (s, 2H), 3.21-3.14 (m, 2H), 1.43 (s, 9H) and 1.21 (d, 12H).
Step F: 2-(4-Fluoro-2,6-diisopropylphenyl)acetic acid
(73) ##STR00060##
(74) To a solution of tert-butyl 2-(4-fluoro-2,6-diisopropylphenyl)acetate (12 g, 40.76 mmol, 1 eq) in DCM (120 mL) was added TFA (184.80 g, 39.76 eq). The reaction mixture was stirred at 25° C. for 3 hours. Most of the solvents were evaporated under reduced pressure. The residue was diluted with H.sub.2O (300 mL) and the mixture was adjusted to pH 10 with 2M aqueous NaOH solution. The mixture was washed with EtOAc (3×500 mL) and the organic phases were discarded. Then the aqueous layer was adjusted to pH 3 with 1M aqueous HCl solution and extracted with EtOAc (3×500 mL. The combined organic layers were washed with brine (2×200 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to give the title compound (8 g, 82%) as a yellow solid.
(75) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.24 (br s, 1H), 6.91 (d, 2H), 3.78 (s, 2H), 3.16-3.06 (m, 2H) and 1.18 (d, 12H).
Intermediate P1: 1-Isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide
Step A: 1-Isopropyl-3-nitro-1H-pyrazole
(76) ##STR00061##
(77) To a mixture of 3-nitro-1H-pyrazole (30 g, 265.31 mmol, 1 eq) in DMF (300 mL) was added NaH (11.14 g, 278.58 mmol, 60% purity in mineral oil, 1.05 eq) in portions at 0° C. Then the reaction mixture was stirred at 0° C. for 0.5 hour. 2-Bromopropane (39.16 g, 318.37 mmol, 1.2 eq) was added and the resulting mixture was warmed to 25° C. for 12 hours. The reaction mixture was quenched with water (500 mL) and extracted with EtOAc (3×300 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 50:1 to 2:1) to give the title compound (29.2 g, 71%) as a yellow oil.
(78) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.49 (d, 1H), 6.90 (d, 1H), 4.63-4.56 (m, 1H) and 1.58 (d, 6H).
Step B: 1-Isopropyl-1H-pyrazol-3-amine
(79) ##STR00062##
(80) To a solution of 1-isopropyl-3-nitro-1H-pyrazole (29.2 g, 188.20 mmol, 1 eq) in MeOH (400 mL) was added Pd/C (3 g, 10 wt % loading on activated carbon) under N.sub.2. The suspension was degassed in vacuo and purged with H.sub.2 several times. The reaction mixture was stirred at 25° C. for 2 hours under H.sub.2 (30 psi). The reaction mixture was filtered and the filtrate was concentrated in vacuo to give the title compound (15.81 g, 66% yield, 98.2% purity on LCMS) as a brown oil.
(81) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.15 (d, 1H), 5.55 (d, 1H), 4.30-4.20 (m, 1H), 3.61 (s, 2H) and 1.43 (d, 6H).
(82) LCMS: m/z 126.2 (M+H).sup.+ (ES.sup.+).
Step C: 1-Isopropyl-1H-pyrazole-3-sulfonyl chloride
(83) ##STR00063##
(84) To a solution of 1-isopropyl-1H-pyrazol-3-amine (15.8 g, 126.23 mmol, 1 eq) in MeCN (600 mL) at 0° C. was added a solution of HCl (116.57 mL, 11.08 eq, 36 wt % in aqueous solution) in H.sub.2 (50 mL). Then an aqueous solution of NaNO.sub.2 (10.45 g, 151-47 mmol, 1.2 eq) in H.sub.2O (50 mL) was added slowly. The resulting mixture was stirred at 0° C. for 0.75 hour. AcOH (50 mL), CuCl (625 mg, 6.31 mmol, 0.05 eq) and CuCl.sub.2 (8.49 g, 63.11 mmol, 0.5 eq) were added. Then SO.sub.2 gas (15 psi) was bubbled into the mixture at 0° C. for 0.25 hour. The reaction mixture was stirred at 0° C. for 1 hour, and then poured into ice water (500 mL) and extracted with DCM (2×700 mL). The combined organic layers were washed with brine (2×700 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 1:0 to 10:1) to give the title compound (18.36 g, 70%) as a colourless oil.
(85) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.56 (d, 1H), 6.88 (d, 1H), 4.70-4.60 (m, 1H) and 1.59 (d, 6H).
Step D: 1-Isopropyl-N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide
(86) ##STR00064##
(87) To a mixture of 1-isopropyl-1H-pyrazole-3-sulfonyl chloride (17.3 g, 82.91 mmol, 1 eq) and TEA (10.91 g, 107.78 mmol, 1.3 eq) in THF (200 mL) was added bis(4-methoxybenzyl)amine (14.93 g, 58.04 mmol, 0.7 eq). The reaction mixture was stirred at 20° C. for 3 hours, and then poured into water (500 mL) and extracted with DCM (2×500 mL). The combined organic layers were washed with brine (2×500 mL), dried over anhydrous Na.sub.2SO.sub.4 filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 20:1 to 4:1) to give the title compound (21.13 g, 59% yield, 100% purity on LCMS) as a colourless oil.
(88) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.46 (d, 1H), 7.09-7.04 (m, 4H), 6.80-6.74 (m, 4H), 6.65 (d, 1H), 4.62-4.54 (m, 1H), 4.32 (s, 4H), 3.79 (s, 6H) and 1.53 (d, 6H).
(89) LCMS: m/z 452.2 (M+Na).sup.+ (ES.sup.+).
Step E: 5-(3-Hydroxyoxetan-3-yl)-1-isopropyl-N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide
(90) ##STR00065##
(91) To a solution of 1-isopropyl-N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide (12 g, 27.94 mmol, 1 eq) in THF (200 mL) was added dropwise n-BuLi (2.5 M, 12.07 mL, in THF, 1.08 eq) at −78° C. Then the reaction mixture was stirred at −78° C. for 1 hour. A solution of oxetan-3-one (2.07 g, 28.78 mmol, 1.03 eq) in THF (50 mL) was added and the resulting mixture was stirred at −78° C. for 1 hour. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl solution (10 mL), poured into water (500 mL) and extracted with ethyl acetate (2×500 mL). The combined organic layers were washed with brine (2×500 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 10:1 to 2:1) to give the title compound (5-11 g, 35% yield, 96.7% purity on LCMS) as a white solid.
(92) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.05 (d, 4H), 6.88 (s, 1H), 6.84-6.80 (m, 4H), 6.79 (s, 1H), 4.88 (d, 2H), 4.77 (d, 2H), 4.46-4.38 (m, 1H), 4.23 (s, 4H), 3.72 (s, 6H) and 1.36 (d, 6H).
(93) LCMS: m/z 502.3 (M+H).sup.+ (ES.sup.+).
Step F: 1-Isopropyl-N,N-bis(4-methoxybenzyl)-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide
(94) ##STR00066##
(95) To a solution of 5-(3-hydroxyoxetan-3-yl)-1-isopropyl-N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide (9.6 g, 19.14 mmol, 1 eq) in DMF (150 mL) was added portionwise NaH (919 mg, 22.97 mmol, 60 wt % in mineral oil, 1.2 eq) at 0° C. The reaction mixture was stirred at 0° C. for 0.5 hour. Then MeI (10.87 g, 76.56 mmol, 4 eq) was added. The reaction mixture was stirred at 0° C. for 13.5 hours, and then warmed to 20° C. The reaction mixture was quenched with saturated aqueous NH.sub.4Cl solution (10 mL) slowly, poured into water (800 mL) and extracted with ethyl acetate (2×500 mL). The combined organic layers were washed with brine (3×500 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo to give the title compound (9.87 g, 94% yield, 94.3% purity on LCMS) as a white solid.
(96) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.14-7.11 (m, 4H), 6.81-6.77 (m, 4H), 6.60 (s, 1H), 4.91 (d, 2H), 4.80 (d, 2H), 4.36 (s, 4H), 4.32-4.25 (m, 1H), 3.79 (s, 6H), 3.05 (s, 3H) and 1.43 (d, 6H).
(97) LCMS: m/z 516.3 (M+H).sup.+ (ES.sup.+).
Step G: 1-Isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide
(98) ##STR00067##
(99) A solution of 1-isopropyl-N,N-bis(4-methoxybenzyl)-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide (9.8 g, 19.01 mmol, 1 eq) in TFA (40 mL) and DCM (40 mL) was stirred at 16° C. for 20 hours. Then the reaction mixture was concentrated in vacuo. The residue was redissolved in THF (80 mL). Hexane (200 mL) was added to the mixture and some solid was precipitated. The colourless precipitate was collected by filtration, washing with hexane (100 ml) and dried in vacuo to give the title compound (3.5 g, 63% yield, 93.7% purity on LCMS) as a light yellow solid.
(100) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.46 (s, 2H), 6.86 (s, 1H), 4.86-4.82 (m, 4H), 4.30-4.20 (m, 1H), 3.00 (s, 3H) and 1.37 (d, 6H).
(101) LCMS: m/z 276.1 (M+H).sup.+ (ES.sup.+).
Intermediate P2: (R)-1-(2-Hydroxypropyl)-1H-pyrazole-3-sulfonamide
Step A: Lithium 1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-sulfinate
(102) ##STR00068##
(103) To a solution of 1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole (100 g, 657.06 mmol, 1 eq) in THF (1.38 L) was added n-BuLi (2.5 M, 276 mL, in THF, 1.05 eq) slowly while keeping the temperature at −70° C. The reaction mixture was stirred for 1.5 hours. Then SO.sub.2 (15 psi) was bubbled into the mixture for 15 minutes. After the reaction mixture was heated to 25° C., a solid was formed. The reaction mixture was concentrated in vacuo. The residue was triturated with tert-butyl methyl ether (400 mL) and the mixture was filtered. The filter cake was washed with tert-butyl methyl ether (50 mL), n-hexane (50 mL) and dried to afford the title compound (142 g, crude) as a white solid.
(104) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.28 (d, 1H), 6.16 (d, 1H), 5.97 (dd, 1H), 3.92-3.87 (m, 1H), 3.61-3.53 (m, 1H), 2.25-2.18 (m, 1H), 1.98-1.93 (m, 1H), 1.78-1.74 (m, 1H) and 1.52-1.49 (m, 3H).
(105) LCMS: m/z 215 (M-Li).sup.− (ES.sup.−).
Step B: 1-(Tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-sulfonyl chloride
(106) ##STR00069##
(107) To a suspension of lithium 1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-sulfinate (20 g, 90.01 mmol, 1 eq) in DCM (250 mL) was added NCS (12.02 g, 90.01 mmol, 1 eq) while cooling in an ice bath. The reaction mixture was stirred at 0° C. for 2 hours. Then the reaction mixture was quenched with water (100 mL) and partitioned between DCM (300 mL) and water (200 mL). The organic layer was washed with water (200 mL), dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give the title compound (15.8 g, 63.02 mmol, 70%) as a yellow oil, which was used directly in the next step.
Step C: N,N-bis(4-Methoxybenzyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-sulfonamide
(108) ##STR00070##
(109) To a mixture of bis(4-methoxybenzyl)amine (16.01 g, 62.23 mmol, 1.04 eq) and TEA (19.33 g, 190.99 mmol, 3.19 eq) in DCM (300 mL) was added a solution of 1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-sulfonyl chloride (15 g, 59.83 mmol, 1 eq) in DCM (50 mL) at 0° C. The reaction mixture was stirred at 0° C. for 1 hour and then quenched with water (250 mL). The organic layer was washed with water (250 mL), 1M aqueous HCl solution (2×250 mL), water (250 mL), and then dried over anhydrous MgSO.sub.4, filtered, and concentrated in vacuo to give the title compound (25.5 g, 83% yield, 92% purity on LCMS) as a brown oil.
(110) LCMS: m/z 494 (M+Na).sup.+ (ES.sup.+).
Step D: N,N-bis(4-Methoxybenzyl)-1H-pyrazole-5-sulfonamide
(111) ##STR00071##
(112) To a solution of N,N-bis(4-methoxybenzyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-sulfonamide (25 g, 53.01 mmol, 1 eq) in THF (183 mL) and MeOH (37 mL) was added a 1M aqueous HCl solution (18.29 mL). The reaction mixture was stirred at 25° C. for 1 hour. Then the reaction mixture was concentrated in vacuo and the residue was partitioned between DCM (200 mL) and H.sub.2O (100 mL). The organic layer was washed with brine (100 mL), dried over anhydrous MgSO.sub.4, filtered and concentrated in vacuo. The residue was triturated with tert-butyl methyl ether (100 mL), filtered and dried to give the title compound (12.2 g, 8% yield, 97% purity on LCMS) as a white solid.
(113) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 13.82-13.70 (br s, 1H), 7.92 (d, 1H), 7.07-7.01 (m, 4H), 6.78-6.75 (m, 4H), 6.61 (d, 1H), 4.34 (s, 4H) and 3.80 (s, 6H).
(114) LCMS: m/z 410 (M+Na).sup.+ (ES.sup.+).
Step E: (R)-1-(2-Hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide
(115) ##STR00072##
(116) To a mixture of N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide (20 g, 51.62 mmol, 1 eq) and (R)-2-methyloxirane (17.99 g, 309.72 mmol, 6 eq) in MeCN (150 mL) was added K.sub.2CO.sub.3 (14.27 g, 103.24 mmol, 2 eq). The reaction mixture was stirred at 50° C. for 20 hours, and then poured into water (500 mL) and extracted with ethyl acetate (2×500 mL). The combined organic layers were washed with brine (2×500 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO.sub.2, petroleum ether:ethyl acetate, 10:1 to 2:1) to give the title compound (12.77 g, 55% yield, 99.5% purity on LCMS) as a light yellow oil.
(117) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.90 (d, 1H), 7.02-6.99 (m, 4H), 6.82-6.80 (m, 4H), 6.70 (d, 1H), 5.02 (d, 1H), 4.18 (s, 4H), 4.14-4.00 (m, 3H), 3.71 (s, 6H) and 1.07-1.06 (d, 3H).
(118) LCMS: m/z 446.2 (M+H).sup.+ (ES.sup.+).
Step F: (R)-1-(2-Hydroxypropyl)-1H-pyrazole-3-sulfonamide
(119) ##STR00073##
(120) A mixture of (R)-1-(2-hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-pyrazole-3-sulfonamide (5.4 g, 12.12 mmol, 1 eq) in TFA (16 mL) and DCM (32 mL) was stirred at 30° C. for 16 hours. Then the reaction mixture was poured into MeOH (100 mL) and the resulting mixture was stirred for 30 minutes. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was dissolved in MeOH (100 mL) and the resulting mixture was adjusted to pH 8 by treating with resin (Amberlyst® A-21, ion exchange resin). The mixture was filtered and the filtrate was concentrated in vacuo to give the title compound (2.38 g, crude) as a light yellow oil.
(121) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.80-7.79 (d, 1H), 7.36 (s, 2H), 6.56-6.55 (d, 1H), 4.97 (d, 1H), 4.10-4.03 (m, 2H), 4.02-3.93 (m, 1H) and 1.06-1.05 (d, 3H).
(122) LCMS: m/z 228.1 (M+Na).sup.+ (ES.sup.+).
SYNTHESIS OF EXAMPLES
Example 1: 2-(1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)-N-((1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazol-3-yl)sulfonyl)acetamide
(123) ##STR00074##
(124) Et.sub.3N (0.67 mL, 4.80 mmol) and 1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide (Intermediate P1) (84 mg, 0.31 mmol) were added to a solution of 2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)acetyl chloride (60 mg, 0.26 mmol) in DCM (5 mL). The mixture was stirred for 48 hours at room temperature and then concentrated. Purification by column chromatography (SiO.sub.2, 0-6% MeOH in DCM) afforded the title compound.
(125) The product was recrystallized from a heptane/DCM mixture, washed with pentane and dried in vacuo to afford the title compound (48 mg, 39%) as a white solid.
(126) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.91 (s, 1H), 7.09 (s, 1H), 6.96 (s, 1H), 4.96-4.81 (m, 4H), 4.34 (m, 1H), 3.63 (s, 2H), 3.06 (s, 3H), 2.90 (t, 4H), 2.72 (t, 4H), 2.08 (m, 4H), 1.45 (d, 6H).
(127) LCMS: m/z 474 (M+H).sup.+ (ES.sup.+); 472 (M−H).sup.− (ES.sup.−).
Example 2: 2-(5-(2-Cyanopyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)-N-((1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazol-3-yl)sulfonyl)acetamide, potassium Salt
(128) ##STR00075##
(129) To a solution of 1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide (Intermediate P1) (49 mg, 0.18 mmol, 1.8 eq) in anhydrous tetrahydrofuran (2 mL) was added potassium tert-butoxide (20 mg, 0.18 mmol, 1.8 eq). The suspension was cooled in an ice bath. A solution of 2-(5-(2-cyanopyridin-4-yl)-2,3-dihydro-H-inden-4-yl)acetyl chloride (Intermediate A1) (30 mg, 0.10 mmol, 1 eq) in anhydrous tetrahydrofuran (2 mL) was added dropwise. After complete addition, the cooling bath was removed and the reaction mixture was allowed to stir at room temperature. After 2 hours, the reaction mixture was concentrated in vacuo. The residue was dissolved in DMSO (0.5 mL) and submitted for purification by reversed phase column chromatography (see “Experimental Methods”, “Purification Method 1”) to afford the title compound (6.0 mg, 0.011 mmol, 11%) as a white solid.
(130) .sup.1H NMR (CD.sub.3OD) δ 8.66 (d, 1H), 7.85 (s, 1H), 7.62 (dd, 1H), 7.21 (d, 1H), 7.04 (d, 1H), 6.90 (s, 1H), 4.84 (d, 4H), 4.41-4.26 (m, 1H), 3.52 (s, 2H), 3.05 (s, 3H), 2.94 (t, 2H), 2.79 (t, 2H), 2.13-1.99 (m, 2H), 1.42 (dd, 6H).
(131) LCMS: m/z 536 (M+H).sup.+ (ES.sup.+).
Example 3: N-((1-Cyclopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazol-3-yl)sulfonyl)-2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)acetamide
(132) ##STR00076##
(133) Prepared as described for 2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)-N-((1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazol-3-yl)sulfonyl)acetamide (Example 1) using 1-cyclopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide (78 mg, 0.28 mmol) and 2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)acetyl chloride (80 mg, 0.34 mmol) to afford the title compound (71 mg, 53%) as a white solid.
(134) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.82 (s, 1H), 7.09 (s, 1H), 7.00 (s, 1H), 4.93 (s, 4H), 3.67-3.54 (m, 3H), 3.09 (s, 3H), 2.91 (t, 4H), 2.71 (t, 4H), 2.09 (m, 4H), 1.28 (m, 2H), 1.02 (m, 2H).
(135) LCMS: m/z 472 (M+H).sup.+ (ES.sup.+); 470 (M−H).sup.− (ES.sup.−).
Example 4: 2-(1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)-N-((1-(2-methoxy-2-methylpropyl)-1H-pyrazol-3-yl)sulfonyl)acetamide
(136) ##STR00077##
(137) Triethylamine (0.67 mL, 4.80 mmol) and 1-(2-methoxy-2-methylpropyl)-1H-pyrazole-3-sulfonamide (60 mg, 0.26 mmol) were added to a solution of 2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)acetyl chloride (60 mg, 0.26 mmol) in DCM (5 mL). The mixture was stirred for 48 hours at room temperature and then concentrated. Purification by column chromatography (SiO.sub.2, 0-6% MeOH in DCM) afforded the title compound (67 mg, 60%) as a white solid.
(138) .sup.1H NMR (300 MHz, CDCl.sub.3) δ 7.83 (s, 1H), 7.59 (d, 1H), 7.07 (s, 1H), 6.93 (d, 1H), 4.16 (s, 2H), 3.60 (s, 2H), 3.22 (s, 3H), 2.89 (t, 4H), 2.71 (t, 4H), 2.07 (m, 4H), 1.11 (s, 6H).
(139) LCMS: m/z 432 (M+H).sup.+ (ES.sup.+); 430 (M−H).sup.− (ES.sup.−).
Example 5: 2-(4-Fluoro-2,6-diisopropylphenyl)-N-((1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazol-3-yl)sulfonyl)acetamide
(140) ##STR00078##
(141) To a mixture of 1-isopropyl-5-(3-methoxyoxetan-3-yl)-1H-pyrazole-3-sulfonamide (Intermediate P1) (69 mg, 251.79 μmol, 1 eq) and 2-(4-fluoro-2,6-diisopropylphenyl)acetic acid (Intermediate A2) (60 mg, 251.79 μmol, 1 eq) in DMF (2 mL) were added EDC (97 mg, 503.57 μmol, 2 eq) and DMAP (62 mg, 503.57 μmol, 2 eq) in one portion at 16° C. The reaction mixture was stirred at 16° C. for 1 hour, and then poured into saturated aqueous citric acid solution (10 mL) and extracted with DCM (2×10 mL). The combined organic layers were washed with brine (2×10 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by prep-HPLC (Column: Waters Xbridge C18, 150 mm*25 mm*5 μm; mobile phase [A: water (0.05% ammonium hydroxide v/v); B: MeCN]; B %: 16%-36%, 10 min) to give the title compound (34 mg, 27% yield, 100% purity on LCMS) as a white solid.
(142) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 6.93 (s, 1H), 6.85 (d, 2H), 4.81-4.77 (m, 4H), 4.26-4.19 (m, 1H), 3.69 (s, 2H), 2.97-2.93 (m, 5H), 1.34 (d, 6H) and 1.04 (d, 12H). LCMS: m/z 496.2 (M+H).sup.+ (ES.sup.+).
Example 6: (R)-2-(4-Fluoro-2,6-diisopropylphenyl)-N-((1-(2-hydroxypropyl)-1H-pyrazol-3-yl)sulfonyl)acetamide
Step A: (R)-1-(2-((tert-Butyldimethylsilyl)oxy)propyl)-1H-pyrazole-3-sulfonamide
(143) ##STR00079##
(144) To a solution of (R)-1-(2-hydroxypropyl)-1H-pyrazole-3-sulfonamide (Intermediate P2) (200 mg, 974.50 μmol, 1 eq) and imidazole (199 mg, 2.92 mmol, 3 eq) in DMF (2 mL) was added TBSCl (154 mg, 1.02 mmol, 1.05 eq) at 0° C. The mixture was stirred at 20° C. for 2 hours, and then poured into water (10 mL) and extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by reversed phase flash chromatography (0.1% NH.sub.3.Math.H.sub.2O-MeCN) to give the title compound (120 mg, 30% yield, 77.5% purity on LCMS) as a white solid.
(145) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.46 (d, 1H), 6.70 (d, 1H), 5.04 (s, 2H), 4.25-4.20 (m, 2H), 4.03-3.99 (m, 1H), 1.29 (d, 3H), 0.82 (s, 9H), −0.03 (s, 3H), −0.22 (s, 3H).
(146) LCMS: m/z 320.1 (M+H).sup.+ (ES.sup.+).
Step B: (R)—N-((1-(2-((tert-Butyldimethylsilyl)oxy)propyl)-1H-pyrazol-3-yl)sulfonyl)-2-(4-fluoro-2,6-diisopropylphenyl)acetamide
(147) ##STR00080##
(148) To a mixture of 2-(4-fluoro-2,6-diisopropylphenyl)acetic acid (Intermediate A2) (90 mg, 375.59 μmol, 1 eq), EDC (144 mg, 751.19 μmol, 2 eq) and DMAP (92 mg, 751.19 μmol, 2 eq) in DMF (2 mL) was added (R)-1-(2-((tert-butyldimethylsilyl)oxy)propyl)-1H-pyrazole-3-sulfonamide (120 mg, 375.59 μmol, 1 eq) at 20° C. The reaction mixture was stirred at 20° C. for 2 hours, and then poured into 0.5 M aqueous HCl solution (20 mL). The mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the title compound (150 mg, crude) as a white solid.
(149) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 7.43 (d, 1H), 6.82 (d, 2H), 6.67 (d, 1H), 4.12-4.01 (m, 1H), 3.99-3.87 (m, 2H), 3.76 (s, 2H), 3.04-2.89 (m, 2H), 1.27-1.07 (m, 15H), 0.84 (s, 9H), −0.04 (s, 3H) and −0.18 (s, 3H).
(150) LCMS: m/z 540.3 (M+H).sup.+ (ES.sup.+).
Step C: (R)-2-(4-Fluoro-2,6-diisopropylphenyl)-N-((1-(2-hydroxypropyl)-1H-pyrazol-3-yl)sulfonyl)acetamide
(151) ##STR00081##
(152) A solution of (R)—N-((1-(2-((tert-butyldimethylsilyl)oxy)propyl)-1H-pyrazol-3-yl)sulfonyl)-2-(4-fluoro-2,6-diisopropylphenyl)acetamide (150 mg, 277.89 μmol, 1 eq) in HCl/dioxane (5 mL, 4M) was stirred at 20° C. for 2 hours. The mixture was concentrated in vacuo. The residue was purified by prep-HPLC (Column: Phenomenex Synergi C18, 150 mm*25 mm*10 μm; mobile phase [A: water (0.225% formic acid); B: MeCN]; B %: 40%-70%, 2 min) to give the title compound (35.91 mg, 30% yield, 99.4% purity on LCMS) as a white solid.
(153) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 12.34 (br s, 1H), 7.83 (s, 1H), 6.87 (d, 2H), 6.69 (s, 1H), 5.00 (d, 2H), 4.12-4.05 (m, 2H), 3.98-3.92 (m, 1H), 3.74 (s, 2H), 2.92-2.88 (m, 2H) and 1.04-1.01 (m, 15H).
(154) LCMS: m/z 426.1 (M+H).sup.+ (ES.sup.+).
(155) SFC: retention time: 0.770 min. ee value: 100%.
(156) The compounds of examples 7-9 were synthesised by methods analogous to those outlined above.
(157) TABLE-US-00007 TABLE 1 .sup.1H NMR and MS data 1H NMR Ex Structure and Name spectrum MS MW 7
(158) Further compounds of the invention may be synthesised by methods analogous to those outlined above.
EXAMPLES—BIOLOGICAL STUDIES
(159) NLRP3 and Pyroptosis
(160) It is well established that the activation of NLRP3 leads to cell pyroptosis and this feature plays an important part in the manifestation of clinical disease (Yan-gang Liu et al., Cell Death & Disease, 2017, 8(2), e2579; Alexander Wree et al., Hepatology, 2014, 59(3), 898-910; Alex Baldwin et al., Journal of Medicinal Chemistry, 2016, 59(5), 1691-1710; Ema Ozaki et al., Journal of Inflammation Research, 2015, 8, 15-27; Zhen Xie & Gang Zhao, Neuroimmunology Neuroinflammation, 2014, 1(2), 60-65; Mattia Cocco et al., Journal of Medicinal Chemistry, 2014, 57(24), 10366-10382; T. Satoh et al., Cell Death & Disease, 2013, 4, e644). Therefore, it is anticipated that inhibitors of NLRP3 will block pyroptosis, as well as the release of pro-inflammatory cytokines (e.g. IL-1β) from the cell.
(161) THP-1 Cells: Culture and Preparation
(162) THP-1 cells (ATCC #TIB-202) were grown in RPMI containing L-glutamine (Gibco #11835) supplemented with 1 mM sodium pyruvate (Sigma #S8636) and penicillin (100 units/ml)/streptomycin (0.1 mg/ml) (Sigma #P4333) in 10% Fetal Bovine Serum (FBS) (Sigma #F0804). The cells were routinely passaged and grown to confluency (˜10.sup.6 cells/ml). On the day of the experiment, THP-1 cells were harvested and resuspended into RPMI medium (without FBS). The cells were then counted and viability (>90%) checked by Trypan blue (Sigma #T8154). Appropriate dilutions were made to give a concentration of 625,000 cells/ml. To this diluted cell solution was added LPS (Sigma #L4524) to give a 1 μg/ml Final Assay Concentration (FAC). 40 μl of the final preparation was aliquoted into each well of a 96-well plate. The plate thus prepared was used for compound screening.
(163) THP-1 Cells Pyroptosis Assay
(164) The following method step-by-step assay was followed for compound screening. 1. Seed THP-1 cells (25,000 cells/well) containing 1.0 μg/ml LPS in 40 μl of RPMI medium (without FBS) in 96-well, black walled, clear bottom cell culture plates coated with poly-D-lysine (VWR #734-0317) 2. Add 5 μl compound (8 points half-log dilution, with 10 μM top dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells 3. Incubate for 3 hrs at 37° C., 5% CO.sub.2 4. Add 5 μl nigericin (Sigma #N7143) (FAC 5 μM) to all wells 5. Incubate for 1 hr at 37° C., 5% CO.sub.2 6. At the end of the incubation period, spin plates at 300×g for 3 mins and remove supernatant 7. Then add 50 μl of resazurin (Sigma #R7017) (FAC 100 μM resazurin in RPMI medium without FBS) and incubate plates for a further 1-2 hrs at 37° C. and 5% CO.sub.2 8. Plates were read in an Envision reader at Ex 560 nm and Em 590 nm 9. IC.sub.50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)
(165) 96-Well Plate Map
(166) TABLE-US-00008 1 2 3 4 5 6 7 8 9 10 11 12 A High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low B High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low C High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low D High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low E High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low F High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low G High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low H High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low High MCC950(10 uM) Compound 8-point half-log dilution Low Drug free control
(167) The results of the pyroptosis assay are summarised in Table 2 below as THP IC.sub.50.
(168) TABLE-US-00009 TABLE 2 NLRP3 inhibitory activity (≤5 μM = ‘++’, ≤10 μM = ‘+’). Example THP No IC.sub.50 1 ++ 2 + 3 ++ 4 ++ 5 ++ 6 ++ 7 ++ 8 ++ 9 ++
(169) As is evident from the results presented in Table 2, surprisingly in spite of the structural differences versus the prior art compounds, the compounds of the invention show high levels of NLRP3 inhibitory activity.
(170) It will be understood that the present invention has been described above by way of example only. The examples are not intended to limit the scope of the invention. Various modifications and embodiments can be made without departing from the scope and spirit of the invention, which is defined by the following claims only.