SULFONAMIDE DERIVATES AS NLRP3 INHIBITORS

Abstract

The present invention relates to compounds of formula (I): (I) wherein Q is selected from O or S; R.sup.1 is an optionally substituted hydrocarbyl group which may optionally include one or more heteroatoms N, O or S in its carbon skeleton, provided that the hydrocarbyl group includes at least one heteroatom N, O or S in its carbon skeleton or is substituted with a substituent comprising at least one heteroatom N, O or S, and provided that the atom of R.sup.1 which is attached to the sulfur atom of the remainder of the molecule is not a ring atom of a cyclic group; R.sup.2 is an ,-substituted cyclic group which may optionally be further substituted; R.sup.3 and R.sup.4 are each independently hydrogen, halogen, OH, NH.sub.2, CN, R.sup.5, OR.sup.5, NHRs 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, optionally substituted cyclic group; and R.sup.5 is independently optionally substituted C.sub.1-C.sub.4 alkyl. 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 NLRP.sub.3.

##STR00001##

Claims

1. A compound of formula (I): ##STR00070## or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: Q is selected from O or S; R.sup.1 is 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, provided that the hydrocarbyl group includes at least one heteroatom N, O or S in its carbon skeleton or is substituted with a substituent comprising at least one heteroatom N, O or S, and provided that the atom of R.sup.1 which is attached to the sulfur atom of the remainder of the molecule is not a ring atom of a cyclic group; R.sup.2 is a cyclic group substituted at the and 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.sup.5, NHR.sup.5 or N(R.sup.5).sub.2; R.sup.4 is hydrogen, halogen, OH, NH.sub.2, CN, R.sup.5, OR.sup.5, NHR.sup.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; and R.sup.5 is independently optionally substituted C.sub.1-C.sub.4 alkyl.

2. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, wherein R.sup.1 is R.sup.21-L-, wherein: L is a saturated or unsaturated C.sub.1-C.sub.12 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.21 is NR.sup.22R.sup.23, OR.sup.24, SR.sup.25, (CNR.sup.26)R.sup.27, (CO)R.sup.28, (CS)R.sup.29, CN or N.sub.3; R.sup.22, R.sup.23, R.sup.24, R.sup.25, R.sup.26, R.sup.27, R.sup.28 and R.sup.29 are each independently hydrogen or a saturated or unsaturated C.sub.1-C.sub.10 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; wherein optionally L and R.sup.22, or L and R.sup.23, or R.sup.22 and R.sup.23 together with the nitrogen 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; wherein optionally L and R.sup.24 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; wherein optionally L and R.sup.25 together with the sulfur 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; wherein optionally L and R.sup.26, or L and R.sup.27, or R.sup.26 and R.sup.27 together with the (CN) group 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; wherein optionally L and R.sup.28 together with the (CO) group 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; and wherein optionally L and R.sup.29 together with the (CS) group 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; provided that the atom of L which is attached to the sulfur atom of the remainder of the molecule is not a ring atom of a cyclic group.

3. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, wherein R.sup.1 is a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl group, wherein the C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl group is substituted with one, two or three substituents independently selected from OR, SR, NHR, N(R).sub.2 or a 3- to 7-membered heterocyclic group, wherein the heterocyclic group itself is optionally substituted with C.sub.1-C.sub.6 alkyl, and wherein R is hydrogen, C.sub.1-C.sub.6 alkyl or C.sub.1-C.sub.6 haloalkyl, wherein the C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl group is optionally further substituted with one, two or three substituents independently selected from halo, CN, NO.sub.2, N.sub.3, or oxo (O), and wherein the C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl or C.sub.2-C.sub.6 alkynyl group optionally includes one or two heteroatoms N, O or S in its carbon skeleton.

4. The compound or a pharmaceutically acceptable salt, solvate or prodrug 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 and positions, and wherein R.sup.2 may optionally be further substituted.

5. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 4, 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 , positions and a second cycloalkyl, cycloalkenyl, non-aromatic heterocyclic, aryl or heteroaryl ring is fused to the aryl or heteroaryl group across the , positions, and wherein R.sup.2 may optionally be further substituted.

6. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, wherein R.sup.2 is a cyclic group substituted at the -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 -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 -position and may optionally be further substituted.

7. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, wherein R.sup.3 and R.sup.4 are hydrogen.

8. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, wherein Q is O.

9. The compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, wherein the compound is selected from the group consisting of: ##STR00071## ##STR00072## ##STR00073## ##STR00074## ##STR00075##

10. (canceled)

11. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1, and a pharmaceutically acceptable excipient.

12. (canceled)

13. A method of treating or preventing a disease, disorder or condition in a subject, the method comprising the step of administering an effective amount of the compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as claimed in claim 1 to the subject, thereby treating or preventing the disease, disorder or condition, optionally wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

14. The method as claimed in claim 13, 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.

15. The method as claimed in claim 13, 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).

16. (canceled)

17. The method as claimed in claim 13, 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 a pharmaceutically acceptable salt, solvate or prodrug 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 a pharmaceutically acceptable salt, solvate or prodrug 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

EXAMPLESCOMPOUND SYNTHESIS

[0441] All solvents, reagents and compounds were purchased and used without further purification unless stated otherwise.

Abbreviations

[0442] 2-MeTHF 2-methyltetrahydrofuran [0443] Ac.sub.2O acetic anhydride [0444] AcOH acetic acid [0445] aq aqueous [0446] Boc tert-butyloxycarbonyl [0447] br broad [0448] Cbz carboxybenzyl [0449] CDI 1,1-carbonyl-diimidazole [0450] conc concentrated [0451] d doublet [0452] DABCO 1,4-diazabicyclo[2.2.2]octane [0453] DAST diethylaminosulfur trifluoride [0454] DCE 1,2-dichloroethane, also called ethylene dichloride [0455] DCM dichloromethane [0456] DIPEA N,N-diisopropylethylamine, also called Hnig's base [0457] DMA dimethylacetamide [0458] DMAP 4-dimethylaminopyridine, also called N,N-dimethylpyridin-4-amine [0459] DME dimethoxyethane [0460] DMF N,N-dimethylformamide [0461] DMSO dimethyl sulfoxide [0462] EDC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [0463] eq or equiv equivalent [0464] (ES.sup.+) electrospray ionization, positive mode [0465] Et ethyl [0466] EtOAc ethyl acetate [0467] EtOH ethanol [0468] h hour(s) [0469] HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate [0470] HPLC high performance liquid chromatography [0471] LC liquid chromatography [0472] m multiplet [0473] m-CPBA 3-chloroperoxybenzoic acid [0474] Me methyl [0475] MeCN acetonitrile [0476] MeOH methanol [0477] (M+H).sup.+ protonated molecular ion [0478] MHz megahertz [0479] min minute(s) [0480] MS mass spectrometry [0481] Ms mesyl, also called methanesulfonyl [0482] MsCl mesyl chloride, also called methanesulfonyl chloride [0483] MTBE methyl tert-butyl ether, also called tert-butyl methyl ether [0484] m/z mass-to-charge ratio [0485] NaHMDS sodium hexamethyldisilazide, also called sodium bis(trimethylsilyl)amide [0486] NaO.sup.tBu sodium tert-butoxide [0487] NBS 1-bromopyrrolidine-2,5-dione, also called N-bromosuccinimide [0488] NCS 1-chloropyrrolidine-2,5-dione, also called N-chlorosuccinimide [0489] NMP N-methylpyrrolidine [0490] NMR nuclear magnetic resonance (spectroscopy) [0491] Pd(OAc).sub.2 palladium acetate [0492] Pd(dba).sub.2 bis(dibenzylideneacetone) palladium(0) [0493] Pd.sub.2(dba).sub.3 tris(dibenzylideneacetone) dipalladium(0) [0494] Pd(dppf)Cl.sub.2 [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium(II) [0495] PE petroleum ether [0496] Ph phenyl [0497] PMB p-methoxybenzyl, also called 4-methoxybenzyl [0498] prep-HPLC preparative high performance liquid chromatography [0499] prep-TLC preparative thin layer chromatography [0500] PTSA p-toluenesulfonic acid [0501] q quartet [0502] RP reversed phase [0503] RT room temperature [0504] s singlet [0505] sat saturated [0506] SCX solid supported cation exchange (resin) [0507] SEM 2-(trimethylsilyl)ethoxymethyl [0508] sept septuplet [0509] t triplet [0510] T3P propylphosphonic anhydride [0511] TBME tert-butyl methyl ether, also called methyl tert-butyl ether [0512] TEA triethylamine [0513] TFAA 2,2,2-trifluoroacetic acid anhydride [0514] TFA 2,2,2-trifluoroacetic acid [0515] THF tetrahydrofuran [0516] TLC thin layer chromatography [0517] TMSCl trimethylsilyl chloride [0518] wt % weight percent or percent by weight [0519] XantPhos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene [0520] Xphos 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl [0521] XtalFluor-E (diethylamino)difluorosulfonium tetrafluoroborate

Experimental Methods

Analytical Methods

[0522] 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: [0523] 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. [0524] An Agilent MercuryPlus 300 instrument fitted with a 7.05 Tesla magnet from Oxford instruments, 4 nuclei auto-switchable probe and Mercury plus console. [0525] A Bruker Avance III spectrometer at 400 MHz fitted with a BBO 5 mm liquid probe. [0526] A Bruker 400 MHz spectrometer using ICON-NMR, under TopSpin program control. [0527] A Bruker Avance III HD spectrometer at 500 MHz, equipped with a Bruker 5 mm SmartProbe.

[0528] 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 (2.150 mm, 2.5 m).

TABLE-US-00001 Pump flow: 0.6 mL/min UV detection: 215, 238 nm Injection volume: 0.2 L Run time: 4.0 min Column temperature: 35 C. Mass detection: API-ES +ve and ive

[0529] Pump Program:

TABLE-US-00002 Gradient Time (min) % A % B 0.0 80 20 0.5 80 20 2.0 0 100

[0530] 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.H.sub.2O in water (v/v); B: Acetonitrile. Column: Kinetex EVO C18 2.130 mm, 5 m.

Reversed Phase HPLC Conditions for the LCMS Analytical Methods

[0531] Methods 1a and 1b: Waters Xselect CSH C18 XP column (4.630 mm, 2.5 m) at 40 C.; flow rate 2.5-4.5 mL min.sup.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.

[0532] Method 1c: Agilent 1290 series with UV detector and HP 6130 MSD mass detector using Waters XBridge BEH C18 XP column (2.150 mm, 2.5 m) 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.

Reversed Phase HPLC Conditions for the UPLC Analytical Methods

[0533] Methods 2a and 2b: Waters BEH C18 (2.130 mm, 1.7 m) 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.

Purification Method 1

[0534] 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.

Revelis C18 Reversed-Phase 12 g Cartridge

[0535]

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 m

[0536] 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.

Separation Runs:

[0537]

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

[0538] Detection wavelength: 215, 235, 254 and 280 nm. Before each new run, the cartridge was cleaned using the conditioning method.

Purification Method 2

[0539] 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 (10030 mm; 5 m).

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

Pump Program:

[0540]

TABLE-US-00006 Time Flow (ml/min) Flow (ml/min) (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

Purification Method 3 (Acidic Prep)

[0541] Preparative reversed phase HPLC was carried out using a Waters X-Select CSH column C18, 5 m (1950 mm), flow rate 28 mL min.sup.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.

Purification Method 4 (Basic Prep)

[0542] Preparative reversed phase HPLC was carried out using a Waters X-Bridge Prep column C18, 5 m (1950 mm), flow rate 28 mL min.sup.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.

[0543] Alternatively automated reversed phase HPLC column chromatography purification was carried out using:

[0544] (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.

[0545] (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.

[0546] (iii) a TELEDYNE ISCO CombiFlash Rf+150. Detection wavelength: 220 nm and 254 nm and 215 nm.

[0547] (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-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic Acid, Trifluoroacetic Acid Salt

Step A: 2-Bromo-4-fluoro-6-(prop-1-en-2-yl)aniline

[0548] ##STR00034##

[0549] 2,6-Dibromo-4-fluoroaniline (10.0 g, 37.2 mmol), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (6.87 g, 40.9 mmol) and potassium carbonate (15.4 g, 112 mmol) were dissolved in dioxane (8 mL) and water (4 mL) and degassed four times under argon atmosphere. Pd(dppf)Cl.sub.2CH.sub.2Cl.sub.2 (1.52 g, 1.86 mmol) was added and the mixture was refluxed for 48 hours. Water (20 mL) and ethyl acetate (40 mL) were added and the layers were separated. The aqueous layer was extracted with ethyl acetate (30 mL). The combined organic layers were dried over sodium sulfate, evaporated to dryness and subjected to column chromatography (SiO.sub.2, heptanes with 15% ethyl acetate) to yield the title compound (3.5 g, 41%) as a light brown oil.

[0550] .sup.1H NMR (300 MHz, CDCl.sub.3) 7.10 (dd, 1H), 6.76 (dd, 1H), 5.36 (bs, 1H), 5.08 (bs, 1H), 4.05 (bs, 2H), 2.05 (s, 3H).

Step B: 4-Fluoro-2-(2-methoxypyridin-4-yl)-6-(prop-1-en-2-yl)aniline

[0551] ##STR00035##

[0552] 2-Bromo-4-fluoro-6-(prop-1-en-2-yl)aniline (8.56 g, 37.2 mmol) and 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (10.5 g, 44.6 mmol) were dissolved in dioxane (10 mL) under N.sub.2 atmosphere. Potassium carbonate (15.4 g, 112 mmol) in water (10 mL) was added. Pd(dppf)Cl.sub.2CH.sub.2Cl.sub.2 (1.52 g, 1.86 mmol) was added and the mixture was stirred overnight at reflux. The dioxane was largely removed by rotary evaporation. Ethyl acetate (100 mL) was added and the layers were separated. The organic layer was washed with brine, dried over sodium sulfate, filtered and evaporated to dryness to yield the title compound (8.0 g, 83%) as a brown oil.

[0553] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.20 (d, 1H), 7.00 (m, 2H), 6.82 (s, 1H), 6.72 (d, 1H), 5.34 (bs, 1H), 5.09 (bs, 1H), 3.98 (s, 3H), 3.80 (bs, 2H), 2.05 (s, 3H).

[0554] LCMS: m/z 259 (M+H).sup.+ (ES.sup.+).

Step C: 4-Fluoro-2-(2-methoxypyridin-4-yl)-6-(isopropyl)aniline

[0555] ##STR00036##

[0556] 4-Fluoro-2-(2-methoxypyridin-4-yl)-6-(prop-1-en-2-yl)aniline (8.0 g, 31 mmol) was dissolved in methanol (so mL). Pd/C (0.4 g, 0.4 mmol) was added and the mixture was stirred overnight under H.sub.2 atmosphere. The product was filtered over Celite and subjected to column chromatography (SiO.sub.2, heptanes with 15% ethyl acetate) yielding the title compound (7.9 g, 99%) as a colourless oil.

[0557] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.21 (d, 1H), 6.98 (dd, 1H), 6.92 (dd, 1H), 6.82 (s, 1H), 6.70 (dd, 1H), 3.98 (s, 3H), 3.61 (bs, 2H), 2.91 (m, 1H), 1.25 (d, 6H).

[0558] LCMS: m/z 261 (M+H).sup.+ (ES.sup.+).

Step D: 4-(2-Bromo-5-fluoro-3-isopropylphenyl)-2-methoxypyridine

[0559] ##STR00037##

[0560] 4-Fluoro-2-(2-methoxypyridin-4-yl)-6-(isopropyl)aniline (200 mg, 768 mol) in acetonitrile (12 mL) at 0 C. was treated with concentrated HBr (1.3 g) in water (1 mL). Sodium nitrite (58.3 mg, 845 mol) in water (1 mL) was added and the mixture was stirred at 0 C. for 45 minutes. Copper(I) bromide (110 mg, 768 mol) and copper(II) bromide (172 mg, 768 mol) were added and the mixture was allowed to reach room temperature over 2 hours. The mixture was poured into saturated sodium carbonate solution (so mL). The mixture was extracted with DCM (250 mL). The combined organic layers were dried over sodium sulfate and evaporated to dryness in vacuo to yield the title compound (160 mg, 64%) as a brown oil.

[0561] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.20 (d, 1H), 7.02 (dd, 1H), 6.85 (d, 1H), 6.82 (dd, 1H), 6.73 (s, 1H), 3.98 (s, 3H), 3.42 (m, 1H), 1.24 (d, 6H).

[0562] LCMS: m/z 324 (M+H).sup.+ (ES.sup.+).

Step E: (2-(tert-Butoxy)-2-oxoethyl) zinc (II) Bromide

[0563] ##STR00038##

[0564] To a mixture of 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) under N.sub.2 atmosphere. The mixture was refluxed for 1 hour. After cooling to 40 C., tert-butyl 2-bromoacetate (55 g, 281.97 mmol, 1 eq) was added and the mixture was refluxed for 2 hours. The mixture was cooled, decanted and the supernatant was used into the next step without further purification (crude).

Step F: tert-Butyl 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetate

[0565] ##STR00039##

[0566] 4-(2-Bromo-5-fluoro-3-isopropylphenyl)-2-methoxypyridine (3.1 g, 9.6 mmol) was dissolved in THF (25 mL) under N.sub.2 atmosphere. Pd.sub.2dba.sub.3 (chloroform adduct) (0.55 g, 0.53 mmol) and Xphos (0.50 g, 1.1 mmol) were added. (2-(tert-Butoxy)-2-oxoethyl) zinc (II) bromide (5.5 g, 21 mmol) in THF (20 ml) (prepared in step E) was added and the mixture was heated to 80 C. and stirred overnight. Then the mixture was cooled to room temperature, filtered over Celite and evaporated to dryness in vacuo. The crude product was subjected to column chromatography (SiO.sub.2, heptanes with a 0 to 20% gradient of ethyl acetate) yielding the title compound (1.7 g, 48%) as a colourless oil.

[0567] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.19 (d, 1H), 7.03 (dd, 1H), 6.82 (d, 1H), 6.78 (dd, 1H), 6.68 (s, 1H), 3.98 (s, 3H), 3.42 (s, 2H), 3.02 (m, 1H), 1.41 (s, 9H), 1.23 (d, 6H).

[0568] LCMS: m/z 360 (M+H).sup.+ (ES.sup.+).

Step G: 2-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic Acid, Trifluoroacetic Acid Salt

[0569] ##STR00040##

[0570] tert-Butyl 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetate (3.4 g, 9.5 mmol) was dissolved in DCM (20 mL) and TFA (15 g, 10 mL, 0.13 mol) and stirred for 6 hours at room temperature. The mixture was evaporated to dryness, yielding the title compound (3.9 g, 99%) as a colourless oil.

[0571] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.19 (d, 1H), 7.03 (dd, 1H), 6.81 (d, 1H), 6.78 (dd, 1H), 6.68 (s, 1H), 3.98 (s, 3H), 3.59 (s, 2H), 3.02 (m, 1H), 1.23 (d, 6H).

[0572] LCMS: m/z 302 (MH).sup. (ES.sup.).

Intermediate A2: 2-(5-(2-Methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetyl Chloride

Step A: 5-(Benzyloxy)-4-bromo-2,3-dihydro-1H-indene

[0573] ##STR00041##

[0574] 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%).

[0575] .sup.1H NMR (300 MHz, CDCl.sub.3) 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

[0576] ##STR00042##

[0577] A solution of 5-(benzyloxy)-4-bromo-2,3-dihydro-1H-indene (1.83 g, 6.04 mmol, 1 eq) in anhydrous tetrahydrofuran (so mL) was bubbled through with nitrogen for 20 minutes. To the degassed solution was added tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct (312 mg, 302 mol, 0.05 eq) and dicyclohexyl(2,4,6-triisopropyl-[1,1-biphenyl]-2-yl)phosphane (288 mg, 604 mol, 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 A1, Step E) 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%).

[0578] .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

[0579] ##STR00043##

[0580] 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 mol, 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 mol, 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%).

[0581] .sup.1H NMR (300 MHz, CDCl.sub.3) 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

[0582] ##STR00044##

[0583] 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 (so 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%).

[0584] .sup.1H NMR (300 MHz, CDCl.sub.3) 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-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetate

[0585] ##STR00045##

[0586] A suspension of 2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (1.08 g, 4.57 mmol, 1 eq), tert-butyl 2-(5-(((trifluoromethyl)sulfonyl)oxy)-2,3-dihydro-1H-inden-4-yl)acetate (1.74 g, 4.57 mmol, 1 eq) and potassium carbonate (1.90 g, 13.7 mmol, 3 eq) in 1,4-dioxane (25 mL) was bubbled through with nitrogen for 20 minutes. After that, [1,1-bis(diphenylphosphino)ferrocene] dichloro palladium (II) (167 mg, 229 mol, 0.05 eq) was added and the reaction mixture was heated to 80 C. After stirring overnight, another batch of [1,1-bis(diphenylphosphino)ferrocene] dichloro palladium (II) (167 mg, 229 mol, 0.05 eq) was added and the temperature of the reaction mixture was increased to 100 C. After 2 more hours of stirring, another batch of [14-bis(diphenylphosphino)ferrocene] dichloro palladium (II) (167 mg, 229 mol, 0.05 eq) was added. After stirring for 20 more hours, another batch of [1,1-bis(diphenylphosphino)ferrocene] dichloro palladium (II) (167 mg, 229 mol, 0.05 eq) was added. After 3 more hours of stirring, the reaction mixture was cooled to room temperature and then filtered. The residue was washed with ethyl acetate and dichloromethane. The filtrates were combined and 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 (358 mg, 1.05 mmol, 23%).

[0587] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.16 (d, 1H), 7.19 (d, 1H), 7.03 (d, 1H), 6.86 (dd, 1H), 6.71 (s, 1H), 3.97 (d, 3H), 3.46 (s, 2H), 2.99 (t, 2H), 2.90 (t, 2H), 2.13 (p, 2H), 1.42 (s, 9H).

Step F: 2-(5-(2-Methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic Acid, Trifluoroacetic Acid Salt

[0588] ##STR00046##

[0589] A solution of tert-butyl 2-(5-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetate (172 mg, 507 mol, 1 eq) in trifluoroacetic acid (1 mL, 13 mmol, 26 eq) was stirred at room temperature. After for 20 hours, more trifluoroacetic acid (0.5 mL, 6.5 mmol, 13 eq) was added. After 2 more hours, the solution was concentrated in vacuo. The crude product was suspended in toluene and then concentrated again; this was performed 3 times to afford the title compound (180 mg, 506 mol, 89%).

[0590] .sup.1H NMR (300 MHz, CD.sub.3OD) 8.23 (dd, 1H), 7.32-7.23 (m, 1H), 7.23-6.99 (m, 3H), 4.07 (s, 3H), 3.59 (s, 2H), 2.97 (dt, 4H), 2.14 (p, 2H).

Step G: 2-(5-(2-Methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetyl Chloride

[0591] ##STR00047##

[0592] To a solution of 2-(5-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetic acid, trifluoroacetic acid salt (219 mg, 0.55 mmol, 1 eq) in anhydrous dichloromethane (10 mL) was added one drop of dimethylformamide and then dropwise oxalyl chloride (145 L, 1.65 mmol, 3 eq) at room temperature. After stirring for 2 hours, the reaction mixture was concentrated in vacuo. The crude product was used in the next step without any purification.

Intermediate A3: 2-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetyl Chloride

[0593] ##STR00048##

[0594] 2-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic acid, TFA salt (Intermediate A1) (61 mg, 0.2 mmol) was stirred in DCM (10 mL) and one drop of dimethylformamide was added followed by the dropwise addition of oxalylchloride (88 L, 1 mmol). The solution was stirred at room temperature for 4 hours and then concentrated thoroughly to afford the title compound (65 mg, 99%) as a yellow oil.

[0595] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.04 (s, 1H), 7.23-7.11 (m, 2H), 7.04 (s, 1H), 6.88-6.75 (m, 1H), 4.40 (s, 3H), 4.08 (s, 2H), 3.17 (m, 1H), 1.27 (m, 6H).

Intermediate A4: 2-(4-Fluoro-2,6-diisopropylphenyl)acetic Acid

Step A: 4-Fluoro-2,6-di(prop-1-en-2-yl)aniline

[0596] ##STR00049##

[0597] 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 (iso mL), and extracted with EtOAc (2150 mL). The combined organic layers were washed with brine (2200 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.

[0598] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.68 (d, 2H), 5.32-5.31 (m, 2H), 5.08 (d, 2H), 3.84 (s, 2H) and 2.07 (d, 6H).

[0599] LCMS: m/z 192.2 (M+H).sup.+ (ES.sup.+).

Step B: 4-Fluoro-2,6-diisopropylaniline

[0600] ##STR00050##

[0601] 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.

[0602] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.76 (d, 2H), 3.56 (s, 2H), 2.99-2.89 (m, 2H) and 1.26 (d, 12H).

[0603] LCMS: m/z 196.2 (M+H).sup.+ (ES.sup.+).

Step C: 2-Bromo-5-fluoro-1,3-diisopropylbenzene

[0604] ##STR00051##

[0605] To a solution of 4-fluoro-2,6-diisopropylaniline (3.7 g, 18.95 mmol, 1 eq) in MeCN (180 mL) was added CuBr (40.08 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.

[0606] .sup.1H NMR (400 MHz, CDCl.sub.3) 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

[0607] ##STR00052##

[0608] A mixture of zinc (80 g) in HCl (i. 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

[0609] ##STR00053##

[0610] 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.

[0611] .sup.1H NMR (400 MHz, CDCl.sub.3) 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

[0612] ##STR00054##

[0613] 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 (3500 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 (3500 mL). The combined organic layers were washed with brine (2200 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.

[0614] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 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: N-Methyl-N-(1-methylpyrrolidin-3-yl)sulfamide

[0615] ##STR00055##

[0616] Chlorosulfonylisocyanate (1.06 g, 7.5 mmol) was dissolved in DCM (5 mL) at 0 C. under N.sub.2 atmosphere. tert-Butanol (0.56 g, 7.5 mmol) in DCM (5 mL) was added dropwise and the mixture was stirred for 30 minutes at 0 C. N,1-dimethylpyrrolidin-3-amine (0.86 g, 7.5 mmol) and triethylamine (0.83 g, 8.3 mmol) in DCM (10 mL) were added dropwise while maintaining the temperature below 8 C. Then the mixture was allowed to reach room temperature and stirred for 48 hours. DCM (30 mL) and water (30 mL) were added. The layers were separated. The water layer was extracted with further DCM (30 mL). The combined organic layers were dried over sodium sulfate and evaporated to dryness. The yellow solid obtained was dissolved in acetonitrile (60 mL) and aqueous 5N HCl (10 mL) was added. The mixture was stirred at 80 C. for 3 hours, and then evaporated to dryness and redissolved in isopropanol (20 mL). Potassium tert-butoxide (0.84 g, 7.5 mmol) was added. The mixture was stirred for 1 hour at room temperature, and then filtered. The filtrate was evaporated to dryness to yield the title compound (1.3 g, 90%) as a light brown oil.

[0617] .sup.1H NMR (300 MHz, DMSO-d.sub.6) 4.22 (m, 1H), 2.51 (s, 3H), 2.38 (t, 1H), 2.17 (s, 3H), 2.12 (t, 1H), 1.90 (m, 1H), 1.65 (m, 3H).

[0618] LCMS: m/z 194 (M+H).sup.+ (ES.sup.+).

Intermediate P2: 3-(Diethylamino)propane-1-sulfonamide

[0619] ##STR00056##

[0620] To a microwave vial containing a solution of 3-chloropropane-1-sulfonamide (boo mg, 3.81 mmol) in acetonitrile (12 mL) was added diethylamine (1.58 mL, 15.2 mmol) and potassium carbonate (526 mg, 3.81 mmol) and the vial was sealed. The mixture was heated at 100 C. in a microwave for 2 hours and subsequently heated by conventional heating at 80 C. overnight. Then the solids were filtered off. The filtrate was concentrated under reduced pressure and purified by column chromatography (SiO.sub.2, 0-20% 3.5M MeOH/NH.sub.3 in DCM) to afford the title compound (406 mg, 55%) as white solid.

[0621] .sup.1H NMR (300 MHz, CDCl.sub.3) 3.22 (t, 2H), 2.69 (m, 6H), 2.15 (m, 2H), 1.10 (m, 6H).

Intermediate P3: ((1R,2R,4S)-2-Hydroxy-7,7-dimethylbicyclo[2.2.1]heptan-1-yl)methanesulfonamide

[0622] ##STR00057##

[0623] To a solution of LiAl.sub.4 (62 mg, 1.62 mmol) in dry THF (4 mL) cooled to 70 C. was added dropwise a solution of ((1R,4S)-7,7-dimethyl-2-oxobicyclo[2.2.1]heptan-1-yl)methanesulfonamide (300 mg, 1.3 mmol) in dry THF (2 mL). The reaction mixture was stirred towards room temperature for 2 hours, and then quenched by carefully adding Glauber's salt followed by some water. The mixture was filtered over a glass filter with EtOAc (10 mL). Water (1 mL) was added to the filtrate. The organic phase was separated, dried (Na.sub.2SO.sub.4), filtered and concentrated to afford the title compound (77 mg, 25%) as a white solid.

[0624] .sup.1H NMR (300 MHz, Chloroform-d) 3.63 (d, 1H), 3.17-3.00 (m, 2H), 1.99-1.82 (m, 1H), 1.82-1.64 (m, 4H), 1.56-1.43 (m, 2H), 108 (s, 3H), 0.84 (s, 3H).

Intermediate P4: 4-Hydroxy-4-methylpentane-1-sulfonamide

[0625] ##STR00058##

[0626] To a solution of methyl 4-sulfamoylbutanoate (750 mg, 4.14 mmol, 1 eq) in anhydrous tetrahydrofuran (15 mL) was added dropwise methyl magnesium bromide (3M solution in diethyl ether, 2.8 mL, 8.28 mmol, 2 eq). After stirring overnight at room temperature, the reaction mixture was quenched with a saturated solution of ammonium chloride. The aqueous solution was extracted four times with ethyl acetate. The organic layers were combined and dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was dissolved in anhydrous tetrahydrofuran (15 mL) and then methyl magnesium bromide (3M solution in diethyl ether, 2.8 mL, 8.28 mmol, 2 eq) was added dropwise. After stirring for six days at room temperature, more methyl magnesium bromide (3M solution in diethyl ether, 2.8 mL, 8.28 mmol, 2 eq) was added. After stirring for one more day, the reaction mixture was quenched with water. The aqueous solution was extracted three times with ethyl acetate. The organic layers were combined, dried over sodium sulfate, filtered and then concentrated in vacuo. The crude product was dissolved in methanol, coated on Agilient hydromatrix (a high purity, inert diatomaceous earth sorbent) and then submitted to normal phase flash chromatography using dichloromethane and ammonia (3.5 M) in methanol to afford the title compound (121 mg, 668 mol, 16%).

[0627] .sup.1H NMR (300 MHz, CDCl.sub.3) 4.57 (s, 2H), 3.20-2.96 (m, 2H), 2.11-1.83 (m, 2H), 1.72-1.54 (m, 2H), 1.23 (s, 6H).

SYNTHESIS OF EXAMPLES

Example 1: 2-(5-(2-Methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)-N(N-methyl-N-(1-methylpyrrolidin-3-yl)sulfamoyl)acetamide, Potassium Salt

[0628] ##STR00059##

[0629] To a suspension of N-methyl-N-(1-methylpyrrolidin-3-yl)sulfamide (Intermediate Pt) (76 mg, 0.39 mmol, 2.5 eq) in anhydrous tetrahydrofuran (2 mL) was added potassium tert-butoxide (47 mg, 0.42 mmol, 2.6 eq). The suspension was stirred for 30 minutes at room temperature. A solution of 2-(5-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetyl chloride (Intermediate A2) (48 mg, 0.16 mmol, 1 eq) in anhydrous tetrahydrofuran (2 mL) was added. After stirring over the weekend at room temperature, the reaction mixture was concentrated in vacuo. The crude product was subjected to preparative HPLC (see Experimental Methods, Purification Method 2) to afford the title compound (2 mg, 4 mol, 3%).

[0630] .sup.1H NMR (300 MHz, CD.sub.3OD) 8.11 (d, 1H), 7.17 (d, 1H), 7.07-6.93 (m, 2H), 6.83 (s, 1H), 4.57 (s, 1H), 3.92 (s, 3H), 3.45 (s, 2H), 3.03-2.80 (m, 8H), 2.74 (d, 3H), 2.46 (s, 3H), 2.44-2.30 (m, 1H), 2.16-2.02 (m, 3H).

[0631] LCMS: m/z 459 (M+H).sup.+ (ES.sup.+).

Example 2: N-((3-(Diethylamino)propyl)sulfonyl)-2-(5-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetamide

[0632] ##STR00060##

[0633] To a suspension of 3-(diethylamino)propane-1-sulfonamide (Intermediate P2) (54 mg, 0.28 mmol, 2 eq) in anhydrous tetrahydrofuran (2 mL) was added potassium tert-butoxide (33 mg, 0.29 mmol, 2.1 eq). The suspension was stirred for 30 minutes at room temperature and then cooled in an ice bath. A solution of 2-(5-(2-methoxypyridin-4-yl)-2,3-dihydro-1H-inden-4-yl)acetyl chloride (Intermediate A2) (42 mg, 0.14 mmol, 1 eq) in anhydrous tetrahydrofuran (2 mL) was added. After complete addition, the ice bath was removed and the reaction mixture was allowed to warm to room temperature. After stirring over the weekend, the reaction mixture was concentrated in vacuo. The crude product 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 (5 mg, 10 mol, 7%).

[0634] .sup.1H NMR (300 MHz, CD.sub.3OD) 8.10 (d, 1H), 7.15 (d, 1H), 7.02-6.95 (m, 2H), 6.86-6.82 (m, 1H), 3.96-3.88 (m, 3H), 3.50 (s, 2H), 3.31-3.24 (m, 4H), 3.18 (q, 4H), 2.94 (dt, 4H), 2.11 (h, 4H), 1.28 (t, 6H).

[0635] LCMS: m/z 460 (M+H).sup.+ (ES.sup.+).

Example 3: N-((3-(Diethylamino)propyl)sulfonyl)-2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetamide, Potassium Salt

[0636] ##STR00061##

[0637] 3-(Diethylamino)propane-1-sulfonamide (Intermediate P2) (78 mg, 0.4 mmol) and KO.sup.tBu (45 mg, 0.4 mmol) were stirred in THF (6 mL) for 45 minutes. A solution of 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetyl chloride (Intermediate A3) (64 mg, 0.2 mmol) in THF (1 mL) was added dropwise. The mixture was stirred overnight at room temperature and then concentrated. The residue was dissolved in DMSO (0.5 mL). The mixture was filtered over cotton wool and submitted for purification by reversed phase column chromatography (see Experimental Methods, Purification Method 1) to afford the title compound (11 mg, 11%) as a white solid.

[0638] .sup.1H NMR (300 MHz, Methanol-d.sub.4) 8.13 (dd, 1H), 7.07 (dd, 1H), 6.98 (dd, 1H), 6.84 (dd, 1H), 6.76 (dd, 1H), 3.93 (s, 3H), 3.48 (s, 2H), 3.21 (dd, 2H), 3.14 (m, 1H), 2.87 (m, 4H), 2.71-2.55 (m, 2H), 2.06-1.89 (m, 2H), 1.24 (d, 6H), 1.11 (t, 6H).

[0639] LCMS: m/z 480 (M+H).sup.+ (ES.sup.+); 478 (MH).sup. (ES.sup.).

Example 4: N-((3-(Diethylamino)propyl)sulfonyl)-2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)acetamide

[0640] ##STR00062##

[0641] A solution of 2-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)acetyl chloride (25.2 mg, 107 mol) in DCM (1.5 mL) was added to a solution of 3-(diethylamino)propane-1-sulfonamide (Intermediate P2) (22.9 mg, 118 mol) and Et.sub.3N (217 mg, 2.15 mmol) in DCM (0.5 mL). The reaction mixture was stirred for 48 hours at room temperature, and then concentrated under reduced pressure. 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 (9 mg, 21%) as a white solid.

[0642] .sup.1H NMR (300 MHz, CD.sub.3OD) 6.90 (s, 1H), 3.51 (s, 2H), 3.24 (t, 4H), 3.13 (q, 4H), 2.85 (q, 8H), 2.06 (dt, 6H), 1.26 (t, 6H).

[0643] LCMS: m/z 393 (M+H).sup.+ (ES.sup.+); 391 (MH).sup. (ES).sup..

Example 5: 2-(4-Fluoro-2,6-diisopropylphenyl)-N(N-methyl-N-(1-methylpyrrolidin-3-yl)sulfamoyl)acetamide

[0644] ##STR00063##

[0645] To a mixture of 2-(4-fluoro-2,6-diisopropylphenyl)acetic acid (Intermediate A4) (160 mg, 672.65 mol, 1 eq) in DMF (5 mL) were added EDC (258 mg, 1.35 mmol, 2 eq), HOBt (91 mg, 672.65 mol, 1 eq) and DMAP (8 mg, 67.26 mol, 0.1 eq) in one portion. Then N-methyl-N-(1-methylpyrrolidin-3-yl)sulfamide (Intermediate P1) (130 mg, 672.65 mol, 1 eq) was added and the reaction mixture was stirred at 25 C. for 12 hours. Then the reaction mixture was directly purified by reversed phase flash chromatography (0.1% NH.sub.3.H.sub.2O-MeCN) and then further purified by prep-HPLC (Column: Phenomenex Synergi C18, 150 mm*25 mm*10 m; mobile phase [A: water (0.1% TFA); B: MeCN]; B %: 25%-55%, 9 min) to give the title compound (9.48 mg, 2% yield, 100% purity on LCMS, TFA salt) as a colourless oil.

[0646] .sup.1H NMR (400 MHz, CDCl.sub.3) 13.37 (br s, 1H), 8.16 (br s, 1H), 6.83 (d, 2H), 5.14-5.12 (m, 1H), 3.79-3.78 (m, 1H), 3.77 (s, 2H), 3.69-3.61 (m, 1H), 3.08-3.05 (m, 1H), 2.91-2.89 (m, 2H), 2.87 (s, 6H), 2.84-2.78 (m, 1H), 2.15-2.05 (m, 2H) and 1.16 (d, 12H).

[0647] LCMS: m/z 414.4 (M+H).sup.+ (ES.sup.+).

Example 6: N-((3-(Diethylamino)propyl)sulfonyl)-2-(4-fluoro-2,6-diisopropylphenyl)acetamide

[0648] ##STR00064##

[0649] To a solution of 3-(diethylamino)propane-1-sulfonamide (Intermediate P2) (100 mg, 514.68 mol, 1 eq) in DMF (2 mL) and DCM (2 mL) were added 2-(4-fluoro-2,6-diisopropylphenyl)acetic acid (Intermediate A4) (123 mg, 514.68 mol, 1 eq), EDC (197 mg, 1.03 mmol, 2 eq) and DMAP (126 mg, 1.03 mmol, 2 eq). The reaction mixture was stirred at 25 C. for 0.5 hour. Then the reaction mixture was purified by reversed phase flash chromatography (0.1% TFA in water-MeCN) and then further purified by prep-HPLC (Column: Xtimate C18, 150 mm*25 mm*5 m; mobile phase [A: water (0.05% ammonium hydroxide v/v), B: MeCN]; B %: 15%-45%, 8 min) to give the title compound (27.83 mg, 13% yield, 100% purity on LCMS) as a white solid.

[0650] .sup.1H NMR (400 MHz, DMSO-d.sub.6) 6.86 (s, 1H), 6.83 (s, 1H), 3.53 (s, 2H), 3.26-3.15 (m, 2H), 3.10 (t, 2H), 2.93-2.84 (m, 6H), 1.89-1.81 (m, 2H), 1.13 (d, 12H) and 1.08 (t, 6H).

[0651] LCMS: m/z 415.2 (M+H).sup.+ (ES.sup.+).

Example 7: N-((((1S)-7,7-Dimethyl-2-oxobicyclo[2.2.1]heptan-1-yl)methyl)sulfonyl)-2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetamide, Potassium Salt

[0652] ##STR00065##

[0653] ((1S)-7,7-Dimethyl-2-oxobicyclo[2.2.1]heptan-1-yl)methanesulfonamide (so mg, 0.22 mmol) and 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic acid (Intermediate A1, non salt form) (130 mg, 0.22 mmol) were stirred in DCM (6 mL). EDC (150 mg, 0.76 mmol) and DMAP (92 mg, 0.76 mmol) were added. The mixture was stirred overnight, diluted with DCM (5 mL) and washed with 1M HCl (aqueous, 3 mL). The organic phase was concentrated. The residue was dissolved in DMSO (0.5 mL) and KO.sup.tBu (97 mg, 0.86 mmol) was added. The mixture was submitted for purification by reversed phase column chromatography (see Experimental Methods, Purification Method 1) to afford the title compound (39 mg, 35%) as a white solid.

[0654] .sup.1H NMR (300 MHz, Methanol-d.sub.4) 8.12 (dd, 1H), 7.07 (dd, 1H), 6.98 (dd, 1H), 6.84 (dd, 1H), 6.76 (dd, 1H), 3.92 (s, 3H), 3.60 (d, 1H), 3.48 (s, 2H), 3.26-3.09 (m, 2H), 2.60-2.44 (m, 1H), 2.35 (dt, 1H), 2.05 (m, 2H), 1.90 (d, 1H), 1.65 (td, 1H), 1.49-1.35 (m, 1H), 1.24 (dd, 6H), 1.06 (s, 3H), 0.85 (s, 3H).

[0655] LCMS: m/z 517 (M+H).sup.+ (ES.sup.+); 515 (MH).sup. (ES.sup.).

Example 8: 2-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)-N(N-methyl-N-(1-methylpyrrolidin-3-yl)sulfamoyl)acetamide, Potassium Salt

[0656] ##STR00066##

[0657] N-Methyl-N-(1-methylpyrrolidin-3-yl)sulfamide (Intermediate P1) (40 mg, 0.21 mmol) and 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic acid (Intermediate A1, non salt form) (120 mg, 0.21 mmol) were stirred in DCM (6 mL). EDC (140 mg, 0.72 mmol) and DMAP (88 mg, 0.72 mmol) were added. The mixture was stirred overnight and concentrated. The residue was dissolved in DMSO (0.6 mL) and KO.sup.tBu (93 mg, 0.83 mmol) was added. The mixture was submitted for purification by reversed phase column chromatography (see Experimental Methods, Purification Method 1) to afford a hygroscopic oil (30 mg). The oil was taken up in DCM and washed with 5% NH.sub.4Cl (aqueous, 2 mL). The organic phase was concentrated and the residue was dissolved in DMSO (0.6 mL) and KO.sup.tBu (93 mg, 0.83 mmol) was added. The mixture was subjected once more to reversed phase column chromatography (see Experimental Methods, Purification Method 1) to afford the title compound (12 mg, 12%) as a white solid.

[0658] .sup.1H NMR (300 MHz, Methanol-d.sub.4) 8.13 (d, 1H), 7.12-7.00 (m, 2H), 6.90 (d, 1H), 6.76 (dd, 1H), 3.93 (s, 3H), 3.42 (s, 2H), 3.19 (dd, 1H), 2.70 (s, 3H), 2.64-2.34 (m, 5H), 2.28 (s, 3H), 1.96 (m, 1H), 1.79 (dd, 1H), 1.25 (dd, 6H).

[0659] LCMS: m/z 479 (M+H).sup.+ (ES.sup.+); 477 (MH).sup. (ES.sup.).

Example 9: 2-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)-N-((((1R,2R,4S)-2-hydroxy-7,7-dimethylbicyclo[2.2.1]heptan-1-yl)methyl)sulfonyl)acetamide, Potassium Salt

[0660] ##STR00067##

[0661] ((1R,2R,4S)-2-Hydroxy-7,7-dimethylbicyclo[2.2.1]heptan-1-yl)methanesulfonamide (Intermediate P3) (39 mg, 0.17 mmol) and 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic acid (Intermediate A1, non salt form) (98 mg, 0.17 mmol) were stirred in DCM (6 mL). EDC (110 mg, 0.59 mmol) and DMAP (71 mg, 0.59 mmol) were added. The mixture was stirred overnight, diluted with DCM (5 mL) and washed with 1M HCl (aqueous, 3 mL). The organic phase was concentrated. The residue was dissolved in DMSO (0.5 mL) and KO.sup.tBu (75 mg, 0.67 mmol) was added. The mixture was submitted for purification by reversed phase column chromatography (see Experimental Methods, Purification Method 1) to afford the title compound (33 mg, 38%) as a white solid.

[0662] .sup.1H NMR (300 MHz, Methanol-d.sub.4) 8.12 (dd, 1H), 7.07 (dd, 1H), 6.98 (dd, 1H), 6.86 (dd, 1H), 6.75 (dd, 1H), 4.10 (dd, 1H), 3.92 (s, 3H), 3.54 (d, 1H), 3.50-3.44 (s, 2H), 3.23 (m, 1H), 3.20 (d, 1H), 1.79-1.61 (m, 4H), 1.47 (m, 2H), 1.24 (dd, 6H), 1.10 (m, 1H), 1.05 (s, 3H), 0.84 (s, 3H).

[0663] LCMS: m/z 519 (M+H).sup.+ (ES.sup.+); 517 (MH).sup. (ES.sup.).

Example 10: 2-(4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)-N-((4-hydroxy-4-methylpentyl)sulfonyl)acetamide, Potassium Salt

[0664] ##STR00068##

[0665] 4-Hydroxy-4-methylpentane-1-sulfonamide (Intermediate P4) (36 mg, 0.20 mmol) and 2-(4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl)acetic acid (Intermediate A1, non salt form) (120 mg, 0.2 mmol) were stirred in DCM (6 mL). EDC (130 mg, 0.7 mmol) and DMAP (85 mg, 0.7 mmol) were added. The mixture was stirred overnight, diluted with DCM (5 mL) and washed with 1M HCl (aqueous, 3 mL). The organic phase was concentrated. The residue was dissolved in DMSO (0.5 mL) and KO.sup.tBu (67 mg, 0.6 mmol) was added. The mixture was submitted for purification by reversed phase column chromatography (see Experimental Methods, Purification Method 1) to afford the title compound (56 mg, 60%) as a white solid.

[0666] .sup.1H NMR (300 MHz, Methanol-d.sub.4) 8.17-8.08 (m, 1H), 7.12-7.04 (m, 1H), 7.04-6.98 (m, 1H), 6.84 (d, 1H), 6.75 (dd, 1H), 3.92 (s, 3H), 3.47 (s, 2H), 3.25-3.11 (m, 3H), 1.83 (tdd, 2H), 1.64-1.49 (m, 2H), 1.24 (d, 6H), 1.18 (s, 6H).

[0667] LCMS: m/z 467 (M+H).sup.+ (ES.sup.+); 465 (MH).sup. (ES.sup.).

[0668] The compound of example 11 was synthesised by methods analogous to those outlined above.

TABLE-US-00007 TABLE 1 UPLC/MS and MS data Retention Ex Structure and Name time (min)* MS MW 11 [00069] 1.69 488.34 (M + H).sup.+ (ES.sup.+); 486.32 (M H).sup. (ES.sup.). 487.6 *: Analytical UPLC/MS was carried out using a Waters Acquity CSH C18, 1.7 m, 2.1 30 mm column eluting with a gradient of 0.1% formic acid in MeCN in 0.1% formic acid in water. The gradient is structured with a starting point of 5% MeCN held from 0.0-0.11 minutes. The gradient from 5-95% occurs between 0.11-2.15 minutes with a flush from 2.15-2.56 minutes. A column re-equilibration to 5% MeCN is from 2.56-2.83 minutes. Temperature: 40 C. Flow rate 0.77 mL min.sup.1. UV spectra of the eluted peaks were measured using an Acquity PDA and mass spectra were recorded using an Acquity QDa detector with ESI pos/neg switching.

[0669] Further compounds of the invention may be synthesised by methods analogous to those outlined above.

ExamplesBiological Studies

NLRP3 and Pyroptosis

[0670] 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-1) from the cell.

THP-1 Cells: Culture and Preparation

[0671] 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 g/ml Final Assay Concentration (FAC). 40 l of the final preparation was aliquoted into each well of a 96-well plate. The plate thus prepared was used for compound screening.

THP-1 Cells Pyroptosis Assay

[0672] The following method step-by-step assay was followed for compound screening. [0673] 1. Seed THP-1 cells (25,000 cells/well) containing 1.0 g/ml LPS in 40 l of RPMI medium (without FBS) in 96-well, black walled, clear bottom cell culture plates coated with poly-D-lysine (VWR #734-0317) [0674] 2. Add 5 l compound (8 points half-log dilution, with 10 M top dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells [0675] 3. Incubate for 3 hrs at 37 C., 5% CO.sub.2 [0676] 4. Add 5 l nigericin (Sigma #N7143) (FAC 5 M) to all wells [0677] 5. Incubate for 1 hr at 37 C., 5% CO.sub.2 [0678] 6. At the end of the incubation period, spin plates at 300g for 3 mins and remove supernatant [0679] 7. Then add 50 l of resazurin (Sigma #R7017) (FAC 100 M resazurin in RPMI medium without FBS) and incubate plates for a further 1-2 hrs at 37 C. and 5% CO.sub.2 [0680] 8. Plates were read in an Envision reader at Ex 560 nm and Em 590 nm [0681] 9. IC.sub.50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)

96-Well Plate Map

[0682]

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) Low Drug free control Compound 8-point half-log dilution

[0683] The results of the pyroptosis assay are summarised in Table 2 below as THP IC.sub.50.

TABLE-US-00009 TABLE 2 NLRP3 inhibitory activity Example THP No IC.sub.50 1 ++ 2 + 3 ++ 4 + 5 ++ 6 + 7 + 8 ++ 9 ++ 10 + 11 +++ (1 M = +++, 5 M = ++, 10 M = +).

[0684] 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.

[0685] 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.