Compounds

Abstract

The present invention relates to sulfonylureas and sulfonylthioureas of formula (I) comprising a 5-membered heteroaryl ring attached to the sulfonyl group, wherein the heteroaryl ring is di-substituted at the 3- and 4-positions relative to the point of attachment of the sulfonyl group, and wherein the group attached to the terminal nitrogen atom of the urea group is either a 1,2,3,5,6,7-hexahydro-s-indacen-4-yl group substituted at the 8-position, or a heteroaryl group substituted at the alpha and alpha positions. 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 NLRP3 inhibition. ##STR00001##

Claims

1. A compound of formula (I): ##STR00045## or a pharmaceutically acceptable salt or solvate thereof, wherein: Q is selected from O or S; V, X and Y are each independently selected from C and N, and W and Z are each independently selected from N, O, S, NH and CH, provided that at least one of V, W, X, Y and Z is N, O, S or NH; R.sup.X and R.sup.Y are each independently a halo, OH, NO.sub.2, NH.sub.2, N.sub.3, SH, SO.sub.2H, SO.sub.2NH, 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 R.sup.X and R.sup.Y together with the atoms X and Y to which they are attached may form a 4- to 12-membered saturated or unsaturated cyclic group fused to ring A, wherein the cyclic group fused to ring A may optionally be substituted; m is 0, 1 or 2; each R.sup.1 is independently a halo, OH, NO.sub.2, NH.sub.2, N.sub.3, SH, SO.sub.2H, SO.sub.2NH, 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 R.sup.X and any R.sup.1 attached to W may together with the atoms W and X to which they are attached form a 4- to 12-membered saturated or unsaturated cyclic group fused to ring A, wherein the cyclic group fused to ring A may optionally be substituted; optionally R.sup.Y and any R.sup.1 attached to Z may together with the atoms Y and Z to which they are attached form a 4- to 12-membered saturated or unsaturated cyclic group fused to ring A, wherein the cyclic group fused to ring A may optionally be substituted; and R.sup.2 is selected from: (i) a group having the formula: ##STR00046## wherein R.sup.20 is a halo, OH, NO.sub.2, NH.sub.2, N.sub.3, SH, SO.sub.2H, SO.sub.2NH, 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; or (ii) a heteroaryl group substituted at the and positions, wherein R.sup.2 may optionally be further substituted.

2. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, wherein ring A is monocyclic.

3. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, wherein at least one of W and Z is O or S.

4. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, wherein any monovalent R.sup.X or R.sup.Y contains from 1 to 8 atoms other than hydrogen.

5. The compound or pharmaceutically acceptable salt- or solvate thereof, as claimed in claim 1, wherein R.sup.X and R.sup.Y are each independently a halo group or a saturated hydrocarbyl group, wherein the saturated hydrocarbyl group may be straight-chained or branched, or be or include cyclic groups, wherein the saturated hydrocarbyl group may optionally be substituted with one or more groups independently selected from halo, CN, OH, NH.sub.2 and oxo (O), and wherein the saturated hydrocarbyl group may optionally include one or two heteroatoms independently selected from N and O in its carbon skeleton.

6. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, wherein each R.sup.1 is independently selected from halo; CN; NO.sub.2; N.sub.3; R.sup.; OH; OR.sup.; R.sup.-halo; R.sup.CN; R.sup.NO.sub.2; R.sup.N.sub.3; R.sup.R.sup.; R.sup.OH; R.sup.OR.sup.; SH; SR; SOR; SO.sub.2H; SO.sub.2R.sup.; SO.sub.2NH.sub.2; SO.sub.2NHR.sup.; SO.sub.2N(R.sup.).sub.2; R.sup.SH; R.sup.SR; R.sup.SOR; R.sup.SO.sub.2H; R.sup.SO.sub.2R.sup.; R.sup.SO.sub.2NH.sub.2; R.sup.SO.sub.2NHR.sup.; R.sup.SO.sub.2N(R.sup.).sub.2; NH.sub.2; NHR.sup.; N(R.sup.).sub.2; R.sup.NH.sub.2; R.sup.NHR.sup.; R.sup.-N(R.sup.).sub.2; CHO; COR.sup.; COOH; COOR.sup.; OCOR.sup.; R.sup.-CHO; R.sup.COR.sup.; R.sup.COOH; R.sup.COOR.sup.; or R.sup.OCOR.sup.; wherein each R.sup. 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.3 groups; and wherein each R.sup. is independently selected from a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl or C.sub.2-C.sub.6 cyclic group, and wherein any R.sup. 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, CCH, oxo (O), or 4- to 6-membered heterocyclic group.

7. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, wherein R.sup.2 has the formula: ##STR00047## wherein R.sup.20 is as defined in claim 1.

8. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 7, wherein R.sup.20 is a fluoro, chloro, bromo or CN group.

9. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, wherein R.sup.2 is a heteroaryl group substituted at the and positions, wherein R.sup.2 may optionally be further substituted.

10. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 9, wherein R.sup.2 has a formula selected from: ##STR00048## wherein: A.sup.1 and A.sup.2 are each independently selected from a straight chain alkylene group, wherein one or two carbon atoms in the backbone of the alkylene group may optionally be replaced by one or two heteroatoms independently selected from nitrogen and oxygen, wherein the alkylene group may optionally be substituted with one or more halo,-OH, CN, O(C.sub.1-C.sub.4 alkyl) or O(C.sub.1-C.sub.4 haloalkyl) groups, and wherein any ring containing A.sup.1 or A.sup.2 is a 5- or 6-membered ring; each R.sup.4 is independently selected from a C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.6 cycloalkyl group, wherein the C.sub.1-C.sub.6 alkyl or C.sub.3-C.sub.6 cycloalkyl group may optionally be substituted with one or more halo, OH, CN, O(C.sub.1-C.sub.4 alkyl) or O(C.sub.1-C.sub.4 haloalkyl) groups; and each R.sup.5 is independently selected from hydrogen or a halo group.

11. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 9, wherein R.sup.2 has the formula: ##STR00049## wherein each R.sup.4 is independently selected from a C.sub.1-C.sub.4 alkyl group.

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

13. The compound or pharmaceutically acceptable salt or solvate thereof, as claimed in claim 1, which is (a) a compound selected from the group consisting of: ##STR00050## ##STR00051## ##STR00052## ##STR00053## or (b) a pharmaceutically acceptable salt or solvate of the selected compound.

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

15. A method of treating by ameliorative or palliative therapy, or of delaying onset 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 by ameliorative or palliative therapy, or delaying onset or reducing risk of the disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

16. The method as claimed in claim 15, 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.

17. The method as claimed in claim 15, 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 (HA2o).

18. A method of inhibiting NLRP3 in a subject, the method 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.

20. The method as claimed in claim 15, wherein the compound or the pharmaceutically acceptable salt or solvate thereof is administered as a pharmaceutical composition further comprising a pharmaceutically acceptable excipient.

21. A prodrug of the compound as claimed in claim 1, or a pharmaceutically acceptable salt or solvate thereof.

22. A method of treating by ameliorative or palliative therapy, or of delaying onset or reducing risk of a disease, disorder or condition in a subject, the method comprising the step of administering an effective amount of the prodrug or the pharmaceutically acceptable salt or solvate thereof, as claimed in claim 21, to the subject, thereby treating by ameliorative or palliative therapy, or delaying onset or reducing risk of the disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

Description

EXAMPLESCOMPOUND 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 DCE 1,2-dichloroethane, also called ethylene dichloride DCM dichloromethane DIPEA N,N-diisopropylethylamine, also called Hunig's base DMA dimethylacetamide DMAP 4-dimethylaminopyridine, also called N,N-dimethylpyridin-4-amine DME dimethoxyethane DMF N,N-dimethylformamide DMSO dimethyl sulfoxide eq or equiv equivalent (ES+) 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)+ 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 NaOtBu 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(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 Sept septuplet sat saturated SCX solid supported cation exchange (resin) t triplet T3P propylphosphonic anhydride TBME tert-butyl methyl ether, also called methyl tert-butyl ether TEA triethylamine TFA 2,2,2-trifluoroacetic acid THF tetrahydrofuran TLC thin layer chromatography wt % weight percent or percent by weight

(3) Experimental Methods

(4) Nuclear Magnetic Resonance

(5) NMR spectra were recorded at 300, 400 or 500 MHz. Spectra were measured at 298 K, unless indicated otherwise, and were referenced relative to the solvent resonance. The chemical shifts are reported in parts per million. Spectra were recorded using one of the following machines: a Bruker Avance III spectrometer at 400 MHz fitted with a BBO 5 mm liquid probe, a Bruker 400 MHz spectrometers using ICON-NMR, under TopSpin program control, a Bruker Avance III HD spectrometer at 500 MHz, equipped with a Bruker 5 mm SmartProbe, 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, or an Agilent MercuryPlus 300 instrument fitted with a 7.05 Tesla magnet from Oxford instruments, 4 nuclei auto-switchable probe and Mercury plus console.

(6) LC-MS

(7) LC-MS Methods: Using SHIMADZU LCMS-2020, Agilent 1200 LC/G1956A MSD and Agilent 1200\G6110A, 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.

(8) Reversed Phase HPLC Conditions for the LCMS Analytical Methods

(9) 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. Gradient information: 0-3.00 min, ramped from 95% water-5% acetonitrile to 5% water-95% acetonitrile; 3.00-3.01 min, held at 5% water-95% acetonitrile, flow rate increased to 4.5 mL min.sup.1; 3.01-3.50 min, held at 5% water-95% acetonitrile; 3.50-3.60 min, returned to 95% water-5% acetonitrile, flow rate reduced to 3.50 mL min.sup.1; 3.60-3.90 min, held at 95% water-5% acetonitrile; 3.90-4.00 min, held at 95% water-5% acetonitrile, flow rate reduced to 2.5 mL min.sup.1.

(10) 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. Gradient information: 0-0.5 min, held at 80% A-20% B; 0.5-2.0 min, ramped from 80% A-20% B to 100% B.

(11) Reversed Phase HPLC Conditions for the UPLC Analytical Methods

(12) 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. Gradient information: 0-0.11 min, held at 95% water-5% acetonitrile, flow rate 0.77 mL min.sup.1; 0.11-2.15 min, ramped from 95% water-5% acetonitrile to 5% water-95% acetonitrile; 2.15-2.49 min, held at 5% water-95% acetonitrile, flow rate 0.77 mL min.sup.1; 2.49-2.56 min, returned to 95% water-5% acetonitrile; 2.56-3.00 min, held at 95% water-5% acetonitrile, flow rate reduced to 0.77 mL min.sup.1.

(13) Preparative Reversed Phase HPLC General Methods

(14) Method 1 (acidic preparation): 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.

(15) Method 2 (basic preparation): 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.

(16) Method 3: Phenomenex Gemini column, 10 m (15025 mm), flow rate=25 mL/min eluting with a water-acetonitrile gradient containing 0.04% NH.sub.3 at pH 10 over 9 minutes using UV detection at 220 and 254 nm. Gradient information: 0-9 minutes, ramped from 8% to 35% acetonitrile; 9-9.2 minutes, ramped from 35% to 100% acetonitrile; 9.2-15.2 minutes, held at 100% acetonitrile.

(17) Method 4: Revelis C18 reversed-phase 12 g cartridge [carbon loading 1.sup.8%; surface area 568 m.sup.2/g; pore diameter 65 Angstrom; pH (5% slurry) 5.1; average particle size 40 m], flow rate=30 mL/min eluting with a water-methanol gradient over 35 minutes using UV detection at 215, 235, 254 and 280 nm. Gradient information: 0-5 minutes, held at 0% methanol; 5-30 minutes, ramped from 0% to 70% methanol; 30-30.1 minutes, ramped from 70% to 100% methanol; 30.1-35 minutes, held at 100% methanol.

Synthesis of Intermediates

Intermediate A1: 8-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene-4-carbonitrile

Step A: 8-Bromo-1,2,3,5,6,7-hexahydro-s-indacen-4-amine

(18) ##STR00036##

(19) To a solution of 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (0.5 g, 2.89 mmol) in DCM (10 mL) at 0 C. was added NBS (0.514 g, 2.89 mmol). The solution was gradually warmed to room temperature and stirred for 12 hours. The reaction mixture was diluted with aq Na.sub.2S.sub.2O.sub.3 (25 mL) and extracted with DCM (220 ml). The combined organic extracts were washed with water (10 mL) and saturated brine (20 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to afford a brown solid. The crude product was purified by chromatography on silica gel (12 g column, 0-10% EtOAc/iso-hexane) to afford the title compound (579 mg, 79%) as a brown solid.

(20) .sup.1H NMR (DMSO-d6) 4.71 (s, 2H), 2.80-2.63 (m, 8H), 2.08-1.91 (m, 4H).

(21) LCMS; m/z 252/254 (M+H).sup.+ (ES.sup.+).

Step B: 8-Amino-1,2,3,5,6,7-hexahydro-s-indacene-4-carbonitrile

(22) ##STR00037##

(23) A solution of 8-bromo-1,2,3,5,6,7-hexahydro-s-indacen-4-amine (579 mg, 2.296 mmol) and dicyanozinc (283 mg, 2.411 mmol) in DMA (10 mL) was degassed for 10 minutes with nitrogen. Then Pd(PPh.sub.3).sub.4 (265 mg, 0.230 mmol) was added and the reaction mixture was heated to 100 C. under N.sub.2 for 18 hours. The reaction mixture was allowed to cool to room temperature and then filtered over Celite eluting with EtOAc (30 mL). The filtrate was washed with sat aq NaHCO.sub.3 (210 mL), water (210 mL) and brine (210 mL), dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The residue was purified by chromatography on silica gel (24 g column 0-40% EtOAc/isohexane) to afford the title compound (96 mg, 20%) as colourless solid.

(24) .sup.1H NMR (DMSO-d6) 5.68 (s, 2H), 2.85 (t, J=7.5 Hz, 4H), 2.64 (t, J=7.4 Hz, 4H), 2.15-1.96 (m, 4H).

(25) LCMS; m/z 199.1 (M+H).sup.+ (ES.sup.+).

Step C: 8-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene-4-carbonitrile

(26) ##STR00038##

(27) Triphosgene (95 mg, 0.320 mmol) was added to a mixture of 8-amino-1,2,3,5,6,7-hexahydro-s-indacene-4-carbonitrile (96 mg, 0.484 mmol) and triethylamine (0.202 ml, 1.453 mmol) in THF (5.5 ml) and heated at reflux for 2 hours. The mixture was concentrated in vacuo and dried azeotropically with toluene (31 ml). The residue was taken up in toluene and filtered through a plug of silica, washing with toluene, and the filtrate was concentrated to afford the title compound (101 mg, 87%) as a colourless solid. A sample was quenched with morpholine in DMSO and was analysed.

(28) LCMS m/z 312.1 (M+H).sup.+ (ES.sup.+).

Intermediate A2: 4-Bromo-8-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene

(29) ##STR00039##

(30) Prepared according to the general procedure of 8-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene-4-carbonitrile (Intermediate A1) from 8-bromo-1,2,3,5,6,7-hexahydro-s-indacen-4-amine (Intermediate A1, Step A) to afford the title compound (197 mg, 97%) as a colourless solid. A sample was quenched with morpholine in DMSO and was analysed.

(31) LCMS m/z 365/367 (M+H).sup.+ (ES.sup.+).

Intermediate P1: 5-(2-Hydroxypropan-2-yl)-1-methyl-1H-pyrazole-3-sulfonamide

(32) ##STR00040##

(33) MeMgCl (3 M in THF, 10.72 mL, 32.2 mmol) was added dropwise over 30 minutes to a stirred ice cold solution of ethyl 1-methyl-3-sulfamoyl-1H-pyrazole-5-carboxylate (1.5 g, 6.43 mmol) in THF (50 mL). Further THF (20 mL) was added to improve stirring, followed by the dropwise addition of further MeMgCl (3 M in THF, 2.14 mL, 6.43 mmol). The reaction was allowed to warm to room temperature and stirred for a further 20 hours. Then the reaction was cooled to 0 C., quenched with sat aq NH.sub.4Cl (30 mL) and extracted with EtOAc (430 mL). The combined organic extracts were washed with brine (30 mL), dried (MgSO.sub.4), filtered and evaporated in vacuo. The residue was purified by chromatography on silica gel (220 g column, 0-20% EtOAc/iso-hexane) to afford the title compound (1.1 g, 77%) as a colourless gum.

(34) .sup.1H NMR (DMSO-d.sub.6) 7.32 (s, 2H), 6.40 (s, 1H), 5.46 (s, 1H), 4.01 (s, 3H), 1.50 (s, 6H).

(35) LCMS m/z 220.0 (M+H).sup.+ (ES.sup.+).

PREPARATION OF EXAMPLES

Example 1: N-((8-Cyano-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5-(2-hydroxypropan-2-yl)-1-methyl-1H-pyrazole-3-sulfonamide

(36) ##STR00041##

(37) 5-(2-Hydroxypropan-2-yl)-1-methyl-1H-pyrazole-3-sulfonamide (Intermediate P1) (33 mg, 0.151 mmol) was dissolved in THF (2 mL) and 60% wt sodium hydride (7 mg, 0.175 mmol) was added and stirred at room temperature for 30 minutes to give a white suspension. 8-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene-4-carbonitrile (Intermediate A1) (37 mg, 0.165 mmol) in THF (2 mL) was added and stirred at room temperature for 4 hours. The reaction mixture was diluted with water (2 mL), concentrated, then redissolved in water (1.5 mL) washed with TBME (23 mL) and the aqueous phase was filtered through a syringe filter. The crude product was purified by chromatography on RP Flash C18 (12 g column, 5-50% MeCN/10 mM ammonium bicarbonate) to afford the title compound (40 mg, 58%) as a colourless solid.

(38) .sup.1H NMR (DMSO-d6) 10.99 (s, 1H), 8.43 (s, 1H), 6.57 (s, 1H), 5.53 (s, 1H), 4.04 (s, 3H), 2.96 (t, J=7.5 Hz, 4H), 2.70 (t, J=7.4 Hz, 4H), 2.06 (p, J=7.3 Hz, 4H), 1.50 (s, 6H).

(39) LCMS; m/z 444.5 (M+H).sup.+ (ES.sup.+).

Example 2: N-((8-Bromo-1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-5-(2-hydroxypropan-2-yl)-1-methyl-1H-pyrazole-3-sulfonamide

(40) ##STR00042##

(41) 5-(2-Hydroxypropan-2-yl)-1-methyl-1H-pyrazole-3-sulfonamide (Intermediate P1) (44 mg, 0.201 mmol) was dissolved in THF (2 ml) and sodium hydride (10 mg, 0.250 mmol) was added and stirred at room temperature for 30 minutes to give a white suspension. 4-Bromo-8-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A2) (62 mg, 0.223 mmol) in THF (2 ml, 24.66 mmol) was added and stirred at room temperature for 2 hours. The reaction mixture was diluted with water (2 ml) and then concentrated and the crude product was purified by chromatography on RP Flash C18 (12 g column, 5-40% MeCN/10 mM ammonium bicarbonate) to afford the title compound (53 mg, 46%) as a colourless solid.

(42) .sup.1H NMR (DMSO-d6) 10.85 (s, 1H), 8.08 (s, 1H), 6.55 (s, 1H), 5.50 (s, 1H), 4.03 (s, 3H), 2.82 (t, J=7.4 Hz, 4H), 2.75 (t, J=7.3 Hz, 4H), 2.06-1.91 (m, 4H), 1.49 (s, 6H).

(43) LCMS; m/z 497/499 (M+H).sup.+ (ES.sup.+).

(44) Other compounds of the invention may be synthesised by methods analogous to those outlined above.

ExamplesBiological Studies

(45) NLRP3 and Pyroptosis

(46) 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.

(47) THP-1 Cells: Culture and Preparation

(48) 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.

(49) THP-1 Cells Pyroptosis Assay

(50) The following method step-by-step assay was followed for compound screening. 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) 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 3. Incubate for 3 hrs at 37 C. in 5% CO.sub.2 4. Add 5 l nigericin (Sigma #N7143) (FAC 5 M) to all wells 5. Incubate for 1 hr at 37 C. and 5% CO.sub.2 6. At the end of the incubation period, spin plates at 300g for 3 mins and remove supernatant 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 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)

(51) 96-Well Plate Map

(52) TABLE-US-00001 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

(53) The results of the pyroptosis assay performed are summarised in Table 1 below as THP IC.sub.50.

(54) Human Whole Blood IL1 Release Assay

(55) For systemic delivery, the ability to inhibit NLRP3 when the compounds are present within the bloodstream is of great importance. For this reason, the NLRP3 inhibitory activity of a number of compounds in human whole blood was investigated in accordance with the following protocol.

(56) Human whole blood in Li-heparin tubes was obtained from healthy donors from a volunteer donor panel. 1. Plate out 80 l of whole blood containing 1 g/ml of LPS in 96-well, clear bottom cell culture plate (Corning #3585) 2. Add 10 l compound (8 points half-log dilution with IoM 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 10 l nigericin (Sigma #N7143) (10 M FAC) 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 300g for 5 mins to pellet cells and remove 20 l of supernatant and add to 96-well v-bottom plates for IL-1 analysis (note: these plates containing the supernatants can be stored at 80 C. to be analysed at a later date) 7. IL-1 was measured according to the manufacturer protocol (Perkin Elmer-AlphaLisa IL-1 Kit AL220F-5000) 8. IC.sub.50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)

(57) The results of the human whole blood assay are summarised in Table 1 below as HWB IC.sub.50.

(58) TABLE-US-00002 TABLE 1 NLRP.sub.3 inhibitory activity Example No Structure THP IC.sub.50 HWB IC.sub.50 1 +++ ND 2 ++++ + [THP IC.sub.50 (0.16 M = ++++, 0.64 M = +++, 2.56 M = ++, 10 M = +, not determined = ND)].

(59) As is evident from the results presented in Table 1, surprisingly in spite of the structural differences versus the prior art compounds, the compounds of the invention show high levels of NLRP3 inhibitory activity in the pyroptosis assay and in the human whole blood assay.

(60) 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.