NOVEL COMPOUNDS AND USES

Abstract

The present invention relates to compounds of formula (I): wherein Q is O or S; R.sup.1 is a cyclic group substituted with at least one group X, wherein R.sup.1 may optionally be further substituted; X is any group comprising a carbonyl group; and R.sup.2 is a cyclic group substituted at the -position, wherein R.sup.2 may optionally be further substituted. 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 dual action of NLRP.sub.3 inhibition and the stimulation of insulin secretion.

##STR00001##

Claims

1. A compound of formula (I): ##STR00133## or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein: Q is O or S; R.sup.1 is a cyclic group substituted with at least one group X, wherein R.sup.1 may optionally be further substituted; X is any group comprising a carbonyl group; and R.sup.2 is a cyclic group substituted at the -position, wherein R.sup.2 may optionally be further substituted.

2. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 1, wherein R.sup.2 is an aryl or a heteroaryl group, wherein the aryl or the heteroaryl group is substituted at the -position, and wherein R.sup.2 may optionally be further substituted.

3. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 2, 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 or 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, wherein R.sup.2 may optionally be further substituted.

4. (canceled)

5. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 1, wherein R.sup.2 is a cyclic group substituted at the and positions, wherein R.sup.2 may optionally be further substituted.

6. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 1, wherein: (i) R.sup.1 is a 6-membered cyclic group substituted with at least one group X, wherein R.sup.1 may optionally be further substituted, (ii) R.sup.1 is a 6-membered cyclic group substituted at the 4-position with at least one group X, wherein X is attached to the 6-membered cyclic group only via the 4-position, and wherein the 6-membered cyclic group may optionally be further substituted; or R.sup.1 is a phenyl group or a 6-membered heteroaryl group, wherein the phenyl group or the 6-membered heteroaryl group is substituted at the 4-position with a group X, wherein X is monovalent, and wherein the phenyl group or the 6-membered heteroaryl group may optionally be further substituted.

7. (canceled)

8. (canceled)

9. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 1, wherein R.sup.1 is a heterocyclic group substituted with at least one group X, wherein R.sup.1 may optionally be further substituted, and wherein optionally the heterocyclic group contains at least one nitrogen atom in the heterocyclic ring.

10. (canceled)

11. The compound or pharmaceutically acceptable salt, solvate or prodrug as of claim 1, wherein X is: ##STR00134## wherein: L is a bond or 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; R.sup.a and R.sup.b together with the atoms to which they are attached may form a cyclic group, or R.sup.a and R.sup.b are each independently selected from hydrogen or an alkyl, alkenyl, alkynyl, Z-, Z-alkylene-, Z-alkenylene- or Z-alkynylene-group, wherein one or more carbon atoms in the backbone of the alkyl, alkenyl, alkynyl, alkylene, alkenylene or alkynylene group may optionally be replaced by one or more heteroatoms N, O or S, wherein each Z- is a cyclic group; and wherein any R.sup.a and R.sup.b may optionally be substituted.

12. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 11, wherein L is 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.

13. The compound or pharmaceutically acceptable salt, solvate or prodrug of claim 1 selected from the group consisting of: ##STR00135## ##STR00136## ##STR00137## ##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142## ##STR00143## ##STR00144## ##STR00145## ##STR00146## ##STR00147## ##STR00148## or a pharmaceutically acceptable salt, solvate or prodrug thereof.

14. (canceled)

15. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt, solvate or prodrug of claim 1, and a pharmaceutically acceptable excipient.

16. A method of treatment or prevention of a disease, disorder or condition, comprising the step of administering to a subject in need thereof an effective amount of a compound, or a pharmaceutically acceptable salt, solvate or prodrug of claim 1, thereby treating or preventing the disease, disorder or condition, optionally wherein (i) the disease, disorder or condition is responsive to NLRP3 inhibition and/or (ii) the disease, disorder or condition is responsive to stimulation of insulin secretion; and/or (iii) the disease, disorder or condition is selected from: an autoinflammatory disease; non-alcoholic steatohepatitis (NASH); or cancer.

17. (canceled)

18. (canceled)

19. (canceled)

20. The method of claim 16, wherein the disease, disorder or condition is responsive to both NLRP3 inhibition and the stimulation of insulin secretion.

21. The method of claim 16, wherein the method is a method of treatment or prevention of two or more comorbid diseases, disorders or conditions, wherein the first disease, disorder or condition is responsive to the stimulation of insulin secretion and the second disease, disorder or condition is responsive to NLRP3 inhibition, and optionally wherein the first disease, disorder or condition is selected from diabetes or prediabetes, and the second disease, disorder or condition is selected from: (i) inflammation; (ii) a cardiovascular disease; (iii) a central nervous system disease; (iv) a renal disease; (v) an ocular disease; (vi) a skin disease; or (vii) a liver disease.

22. (canceled)

23. A method of inhibiting NLRP3 and/or stimulating insulin secretion, the method comprising administering a compound or a pharmaceutically acceptable salt, solvate or prodrug of claim 1, to a subject in need thereof thereby inhibiting NLRP3 and/or stimulating insulin secretion.

24. The method of claim 23, wherein the method is a method of inhibiting NLRP3 and stimulating insulin secretion.

25. A method of treatment or prevention of a disease, disorder or condition, comprising the step of administering to a subject in need thereof an effective amount of a pharmaceutical composition of claim 10, thereby treating or preventing the disease, disorder or condition, optionally wherein: (i) the disease, disorder or condition is responsive to NLRP3 inhibition; and/or (ii) the disease, disorder or condition is responsive to the stimulation of insulin secretion; and/or (iii) the disease, disorder or condition is selected from an autoinflammatory disease, non-alcoholic steatohepatitis (NASH), or cancer.

26. The method of claim 25, wherein the disease, disorder or condition is responsive to both NLRP3 inhibition and the stimulation of insulin secretion.

27. The method of claim 25, wherein the method treats or prevents two or more comorbid diseases, disorders or conditions, wherein the first disease, disorder or condition is responsive to the stimulation of insulin secretion and the second disease, disorder or condition is responsive to NLRP3 inhibition, and optionally wherein the first disease, disorder or condition is selected from diabetes or prediabetes, and the second disease, disorder or condition is selected from: (i) inflammation; (ii) a cardiovascular disease; (iii) a central nervous system disease; (iv) a renal disease; (v) an ocular disease; (vi) a skin disease; (vii) a liver disease; (viii) a lung disease; (ix) a cancer; or (x) a metabolic disease that is not responsive to the stimulation of insulin secretion.

28. A method of inhibiting NLRP3 and/or stimulating insulin secretion, the method comprising administering a pharmaceutical composition of claim 10 to a subject in need thereof thereby inhibiting NLRP3 and/or stimulating insulin secretion.

29. The method of claim 28, wherein the method is a method of inhibiting NLRP3 and stimulating insulin secretion.

Description

FIGURES

[0149] FIG. 1 shows the insulin-secretory activity of compounds of the invention and other comparative compounds.

EXAMPLESCOMPOUND SYNTHESIS

[0150] All solvents, reagents and compounds were purchased and used without further purification unless stated otherwise.

[0151] For Intermediates I-3 to I-17 and Examples 1-22:

[0152] .sup.1H NMR and .sup.13C NMR spectra were recorded using a BrukerAvance 600 MHz spectrometer (operating at 600 MHz for .sup.1H NMR and 151 MHz for .sup.13C NMR) .sup.13C and .sup.1H chemical shifts (), reported in ppm, were internally referenced to tetramethylsilane (TMS). LC-MS analysis was performed using a 0.05% (v/v) formic acid.sub.(aq)/0.05% formic acid in CH.sub.3CN solvent system on a Shimadzu Prominence instrument equipped with an Agilent Eclipse XDB-Phenyl column (3.5100 mm, 3 m) maintained at 40 C., SPD-M20A diode array UV-Vis detector, ELSD-LT II evaporative light scattering detector (ELSD) and LC-MS-2020 mass spectrometer. High resolution mass spectrometry (HRMS) was performed on a Bruker MicroTOF mass spectrometer with electrospray ionisation (ESI). Medium pressure liquid chromatography (MPLC) purification was conducted on a Grace Reveleris X1 using two serial 12 g Grace C18 columns with a 10 mM NH.sub.4HCO.sub.3(aq)/CH.sub.3CN solvent system flowing at 30 mL/min. HPLC purification was performed on a Gilson PLC 2020 instrument using an Agilent Eclipse XDB-Phenyl column (21.2 mm100 mm, 5 m) with 10 mM NH.sub.4HCO.sub.3(aq)/CH.sub.3CN solvent system flowing at 20 mL/min.

[0153] For Intermediate I-23 and Examples 23-31:

[0154] Analytical methods: NMR spectra were run on Bruker 400 MHz spectrometers using ICON-NMR, under TopSpin program control. Spectra were measured at 298 K, unless indicated otherwise, and were referenced relative to the solvent resonance.

[0155] LC-MS Methods: Using SHIMADZU LCMS-2020, Agilent 1200 LC/G1956A MSD and Agilent 1200\G6110A, Agilent 1200 LC & Agilent 6110 MSD.

[0156] Purification Method: Automated reversed phase column chromatography was carried out using a Gilson 281 system driven by Gilson 322 pump, Gilson 156 UV/Vis detection unit and Gilson GX-281 fraction collector. The Phenomenex Gemini column (150 mm25 mm10 um) was conditioned prior to use with CH.sub.3CN (2 min) then brought to 5% water (0.05% ammonia hydroxide vol/vol) in 0.3 min and kept 2 min at 5% water (0.05% ammonia hydroxide v/v). Flow rate=25 mL/min. Separation runs:

TABLE-US-00001 A: water (0.05% ammonia B: Time hydroxide CH.sub.3CN (min) V/V %) (%) 0 95 5 12 65 35 12.2 0 100 14.2 0 100 14.5 95 5

[0157] Detection wavelengths=220 and 254 nm. Before each new run the cartridge was cleaned using the conditioning method.

General Synthetic Methods

[0158] ##STR00053##

[0159] An isocyanate was prepared in situ from the corresponding amine by dissolving di-tert-butyl dicarbonate (1.1 eq.) in tetrahydrofuran (THF) (3 mL/mmol amine), treating with N,N-dimethylpyridin-4-amine (DMAP) (1.1 eq.) and stirring for 5 min at room temperature before adding the amine (1 eq.) and stirring for a further 20 min. Meanwhile a sulfonamide sodium salt was prepared in situ by dissolving the a sulfonamide (1 eq.) in THF (3 mL/mmol), treating with NaH (1.0 eq., 60% oil dispersion) and stirring under reduced pressure until effervescence ceased (ca. 5-10 min). The sulfonamide salt and isocyanate solutions were combined and stirred at room temperature for 15 h under N.sub.2 atmosphere, monitored by LC-MS. Reaction mixtures were concentrated in vacuo, dissolved in the minimum volume 1:1 CH.sub.3CN/dimethylformamide (DMF) and purified via reverse phase MPLC. Typically a four minute aqueous wash was followed by a 15 min 10 mM NH.sub.4HCO.sub.3(aq)/CH.sub.3CN gradient.

R.sup.2NH.sub.2.fwdarw.R.sup.2NCOGENERAL METHOD A1:

[0160] To a solution of R.sup.2 amine intermediate (1 eq.) with or without a base such as triethylamine (1.2 eq.), in an anhydrous aprotic solvent such as tetrahydrofuran or dichloromethane, was added triphosgene (0.4 to 1.1 eq.). The reaction was stirred at ambient temperature or, where necessary, heated at reflux until completion, typically from 2 to 18 h.

R.sup.2NH.sub.2.fwdarw.R.sup.2NCOGENERAL METHOD A2:

[0161] To di-t-butyldicarbonate (1.2-1.4 eq.) in anhydrous acetonitrile or THF was added DMAP (15-100 mol %), after 5 minutes, a solution of R.sup.2 amine intermediate (1.0 eq.) in acetonitrile was added. The reaction mixture was stirred for 30-60 min at room temperature.

##STR00054##

[0162] To R.sup.1 sulfonamide intermediate (1 eq) in anhydrous THF (5 mL/mmol) was added NaH (1 eq) at 0 C. and stirred for 30 min to 2 h, or until completion, at ambient temperature under nitrogen atmosphere. Again cooled to 0 C., R.sup.2 isocyanate (1.0 eq) in THF was added and stirred at ambient temperature until completion, typically 2 to 16 h.

##STR00055##

[0163] To crude R.sup.2 isocyanate (1.0 eq) in anhydrous THF or DCM (5-11 mL/mmol) was added R.sup.1 sulfonamide (1.0 eq) followed by base such as triethylamine, DIPEA, or DBU (1-2 eq) and the reaction mixture stirred at ambient temperature overnight.

##STR00056##

[0164] R.sup.1 sulfonamide (1.0 eq) was dissolved in anhydrous THF under a nitrogen atmosphere. Solid sodium methoxide (1.0 eq) was added in one portion. This mixture was stirred at ambient temperature for 3 h. A solution of the R.sup.2 isocyanate (1.17 eq) in THF was added drop wise. The reaction mixture was stirred at room temperature overnight.

##STR00057##

[0165] Alkyne (1 eq) and azide (1.2 eq), 5 mol % CuSO.sub.4, 10 mol % NaAsc solution in DMSO (500 L) were stirred at room temperature until completion, typically 12 h.

##STR00058##

##STR00059##

##STR00060##

[0166] Other compounds of Formula I wherein R.sup.1 is a pyrazole moiety having a different regiochemistry may be prepared by methods analogous to general methods D1-D3.

Intermediate Synthesis

Intermediate I-3: 5-Methyl-N-(4-sulfamoylphenethyl)pyrazine-2-carboxamide

[0167] ##STR00061##

[0168] N-(4-(N-(Cyclohexylcarbamoyl)sulfamoyl)phenethyl)-5-methylpyrazine-2-carboxamide (250 mg, 0.56 mmol) dissolved in pyridine (8 mL) was treated with phthalic anhydride (83 mg, 0.56 mmol) and DMAP (10 mg, 0.082 mmol), then heated at reflux for 5 h under N.sub.2 atmosphere. The reaction mixture was purified by MPLC affording the titled compound as an amorphous white solid (113 mg, 63%): .sup.1H NMR (600 MHz, DMSO-d.sub.6) 9.03 (d, J=1.4 Hz, 1H), 8.96 (t, J=6.0 Hz, 1H), 8.61 (m, 1H), 7.73 (d, J=8.4 Hz, 2H, 7.42 (d, J=8.4 Hz, 2H), 7.30 (s, 2H), 3.57 (dt, J=7.3, 6.0 Hz, 2H, 2.95 (t, J=7.3 Hz, 2H), 2.58 (s, 3H); .sup.13C NMR (151 MHz, DMSO-d.sub.6) 162.8, 156.7, 143.4 (C-1), 142.7 (C-11), 142.3 (C-14), 141.9 (C-12), 141.9 (C-6), 129.0 (C-4, 5), 125.6, 39.8, 34.6, 21.2; HRMS (ESI-TOF) m/z calcd for C.sub.14H.sub.17N.sub.4O.sub.3S [M+H].sup.+ 321.1016, found 321.1029; LC-MS m/z 321.0 [M+H].sup.+, purity >95% (ELSD).

Intermediate I-4: 5-Methyl-N-(4-sulfamoylphenethyl)isoxazole-3-carboxamide

[0169] ##STR00062##

[0170] 5-Methylisoxazole-3-carboxylic acid (150 mg, 1.18 mmol) dissolved in toluene (2 mL) was treated with DMF (1 drop) and thionyl chloride (1.42 mmol, 103 L) then heated to reflux for 5 h. The solvent was removed in vacuo to afford crude 5-methylisoxazole-3-carbonyl chloride (155 mg, 1.06 mmol) as a brown oil. The crude acid chloride without purification was dissolved in THF (4 mL), treated with Et.sub.3N (155 L, 1.06 mmol) and stirred for 5 min, before adding 4-(2-aminoethyl)benzenesulfonamide (220 mg, 1.10 mmol) and stirring at room temperature for 15 h under N.sub.2 atmosphere. The reaction mixture was concentrated in vacuo and purified by MPLC, affording the titled compound as an amorphous white solid (205 mg, 62%): .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.79 (t, J=5.8 Hz, 1H), 7.74 (d, J=8.3 Hz, 2H), 7.41 (d, J=8.3 Hz, 2H), 7.30 (s, 2H), 6.50 (q, J=0.8 Hz, 1H), 3.49 (m, 2H), 2.91 (t, J=7.2 Hz, 2H), 2.45 (d, J=0.8 Hz, 3H); .sup.13C NMR (151 MHz, DMSO-d.sub.6) 171.0, 158.8, 158.5, 143.3, 142.0, 129.0, 125.6, 101.1, 39.8, 34.3, 11.7; HRMS (ESI-TOF) m/z calcd for C.sub.13H.sub.14N.sub.3O.sub.4S [MH].sup. 308.0711, found 308.0708; LC-MS m/z 308.0 [M+H].sup.+, purity >99% (ELSD).

Intermediate I-5: 4-(2-(7-Methoxy-4,4-dimethyl-1,3-dioxo-3,4-dihydroisoquinolin-2(1H)-yl)ethyl)benzenesulfonamide

[0171] ##STR00063##

[0172] N-(Cyclohexylcarbamoyl)-4-(2-(7-methoxy-4,4-dimethyl-1,3-dioxo-3,4-dihydroisoquinolin-2(1H)-yl)ethyl)benzenesulfonamide (504 mg, 0.96 mmol) dissolved in pyridine (8 mL) was treated with phthalic anhydride (143 mg, 0.97 mmol) and DMAP (11.8 mg, 0.097 mmol), then heated at reflux for 5 h under N.sub.2 atmosphere. The reaction mixture was purified by MPLC, affording the titled compound as an amorphous white solid (291 mg, 77%): .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.72 (d, J=8.5 Hz, 2H), 7.61 (d, J=8.8 Hz, 1H), 7.53 (d, J=2.9 Hz, 1H), 7.40 (d, J=8.5 Hz, 2H), 7.33-7.26 (m, 3H), 4.13 (t, J=7.4 Hz, 2H), 3.84 (s, 3H), 2.93 (t, J=7.4 Hz, 2H), 1.45 (s, 6H); .sup.13C NMR (151 MHz, DMSO-d.sub.6) 176.3, 163.1, 158.0, 142.6, 142.2, 137.2, 129.1, 127.4, 125.6, 124.1, 121.5, 110.5, 55.3, 42.3, 40.4, 32.9, 28.8; HRMS (ESI-TOF) m/z calcd for C.sub.20H.sub.21N.sub.2O.sub.5S [MH].sup. 401.1177, found 401.1174; LC-MS m/z: 403.1 [M+H].sup.+, purity >95% (ELSD).

Intermediate I-9: Methyl 3-sulfamoylbenzoate

[0173] ##STR00064##

[0174] Methyl 3-(chlorosulfonyl)benzoate (1.00 g, 4.26 mmol) was dissolved in anhydrous tetrahydrofuran (15 mL) and the solution was cooled to 0 C. Aqueous ammonia (5.0 mL) was added drop-wise and the mixture stirred at ambient temperature for 2 h. Upon completion the reaction mixture was poured into chilled water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The resulting solid was triturated with pentane to afford the titled compound as a light brown solid (0.75 g, 82%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): =8.40 (s, 1H), 8.19 (d, J=8 Hz, 1H), 8.1 (d, J=8 Hz, 1H), 7.77 (t, J=8 Hz, 1H), 7.6 (s, 2H), 3.92 (s, 3H); m/z 214.0 [MH.sup.+].sup..

Intermediate I-12: N-(3-Sulfamoylphenyl)pent-4-ynamide

[0175] ##STR00065##

[0176] To a solution of pent-4-ynoic acid (0.1 g, 1.02 mmol) and 3-aminobenzenesulfonamide (0.21 g, 1.22 mmol) in dry DMF (5.0 ml) was added HBTU (0.46 g, 1.22 mmol) followed by DIPEA (212 uL, 1.22 mmol). The reaction mixture was stirred at ambient temperature for 2 h, or until completion. The mixture was diluted with EtOAc (30 mL), washed with H.sub.2O (20 mL), brine (20 mL) then the organics dried (MgSO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 100% hexanes eluant to give the titled compound as a to give the titled compound as a pale-yellow solid (0.2 g, 79%). .sup.1H NMR (400 MHz, CD.sub.3OD) =8.22 (dd, J=2.2, 1.7 Hz, 1H), 7.75-7.68 (m, 1H), 7.65-7.58 (m, 1H), 7.51-7.42 (m, 2H), 2.64-2.59 (m, 2H), 2.58-2.54 (m, 2H), 2.32-2.25 (m, 1H). .sup.13C NMR (101 MHz, CD.sub.3OD) =171.3, 143.8, 138.9, 129.2, 122.9, 121.0, 117.1, 82.1, 69.1, 35.4, 14.0.

Intermediate I-13: N-(Prop-2-yn-1-yl)-3-(4-sulfamoylphenyl)propanamide

[0177] ##STR00066##

[0178] To a solution of 3-(4-sulfamoylphenyl)propanoic acid (0.3 g, 1.5 mmol) and propargyl amine (0.11 g, 1.5 mmol) in dry DMF (5.0 ml) was added HBTU (0.74 g, 1.5 mmol) followed by DIPEA (342 uL, 1.22 mmol). The reaction mixture was stirred at RT for 2 h. The reaction was monitored by LCMS and after the completion of reaction, it was diluted with EtOAc (30 mL) washed with H2O (20 mL), brine (20 mL). The organic layer was separated; dried (MgSO.sub.4) and evaporated to give the crude product. The crude product was purified by silica gel column chromatography (1:1, EtOAc:Hexane) to isolate the title compound as a white solid (0.22 g, 63%). .sup.1H NMR (400 MHz, CD.sub.3OD) =7.85 (d, J=7.9 Hz, 2H), 7.38 (d, J=7.9 Hz, 2H), 3.97 (t, J=2.4 Hz, 2H), 3.04 (t, J=7.6 Hz, 2H), 2.55 (t, J=7.6 Hz, 2H), 2.33 (d, J=2.8 Hz, 1H).

Intermediate I-14: 2-(Methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)-N-(4-sulfamoylphenethyl)acetamide

[0179] ##STR00067##

[0180] 2-(Methylamino)acetic acid (0.24 g, 2.75 mmol) and sodium hydrogencarbonate (0.694 g, 8.26 mmol) were dissolved in a mixture of water (10 mL) and MeOH (20 mL). Then, 4-chloro-7-nitrobenzo[c][1,2,5]oxadiazole (0.50 g, 2.50 mmol) was added and the mixture stirred at 60 C. for 2 h. Upon completion of the reaction, volatiles were removed under reduced pressure and the crude residue obtained was purified by column chromatography on silica gel using 0-5% gradient of methanol in dichloromethane to obtain 2-(methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)acetic acid as a brick-red solid (1.10 g, 87%).

[0181] 2-(Methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)acetic acid (1.00 g, 3.96 mmol) was dissolved in anhydrous tetahydrofuran (25 mL) under nitrogen atmosphere and the solution was cooled to 0 C. Diisopropylethylamine (0.76 g, 5.55 mmol) and 1,1-carbonyldiimidazole (0.90 g, 4.75 mmol) were added and the mixture stirred at 50 C. until all of the 2-(methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)acetic acid had reacted. The reaction mixture was then cooled to 0 C., 4-(2-aminoethyl)-benzenesulfonamide (0.95 g, 4.75 mmol) was added and stirred at ambient temperature until completion, typically 6 h. The solvents were removed in vacuo and the residue was purified by reverse phase preparative HPLC to afford the titled compound as a brick-red solid (1.20 g, 70%). LCMS (m/z): 435.4 (M+1).sup.+.

Intermediate I-15: 2-(7-(Dimethylamino)-2-oxo-2H-chromen-4-yl)-N-(4-sulfamoylphenethyl)acetamide

[0182] ##STR00068##

[0183] 2-(7-(dimethylamino)-2-oxo-2H-chromen-4-yl)acetic acid (0.50 g, 2.02 mmol), EDC-HC (0.47 g, 3.03 mmol), HOBt (0.464 g, 3.03 mmol) and N-methylmorpholine (0.409 g, 4.04 mmol) were mixed in anhydrous tetrahydrofuran (5 mL) and stirred at 0 C. for 30 min. 4-(2-Aminoethyl)benzenesulfonamide (0.445 g, 2.224 mmol) was added and stirring continued at ambient temperature for 18 h. Upon completion, the reaction was poured onto chilled water and extracted with ethyl acetate. The solvent was removed in vacuo and the residue was purified by column chromatography on silica gel using a gradient of 0-5% methanol in dichloromethane to give 2-(7-(dimethylamino)-2-oxo-2H-chromen-4-yl)-N-(4-sulfamoylphenethyl)acetamide as a greenish-yellow solid (0.25 g, 29%). LCMS (m/z): 430.2 (M+1).sup.+.

Intermediate 1-16: 3,5,6,7-Tetrahydro-2H-indeno[5,6-b]furan-8-amine

[0184] ##STR00069##

[0185] A solution of 2,3-dihydrobenzofuran-5-carbaldehyde (10 g, 67.6 mmol), malonic acid (10.5 g, 101.35 mmol) and piperidine (0.47 mL, 4.73 mmol, 0.07 eq) was heated in pyridine (60 mL) at 100 C. for 5 h. The reaction mixture was acidified to pH 3 using 1N HCl and the product extracted using 10% IPA/chloroform (2250 mL). The combined organic extracts were washed with water (250 mL), brine (250 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was triturated using diethyl ether to give (E)-3-(2,3-dihydrobenzofuran-5-yl)acrylic acid as a yellow solid (10 g, 78%). .sup.1H NMR (300 MHz, Chloroform-d) =7.73 (d, J=15.9 Hz, 1H), 7.43 (s, 1H), 7.33 (dd, J=8.1, 1.8 Hz, 1H), 6.80 (d, J=8.1 Hz, 1H), 6.29 (d, J=15.9 Hz, 1H), 4.64 (t, J=8.7 Hz, 2H), 3.24 (t, J=8.7 Hz, 2H).

[0186] A solution of (E)-3-(2,3-dihydrobenzofuran-5-yl)acrylic acid (8.0 g, 42.1 mmol) in acetic acid (80 mL) and water (1.0 mL) was treated with 10% palladium on carbon (1.0 g) in two portions. The reaction mixture was stirred under an atmosphere or hydrogen gas (balloon) until completion, typically 4 h. The mixture was diluted using ethyl acetate (100 mL) and filtered through a bed of celite washing through with further ethyl acetate. The solvents were removed in vacuo and the crude residue azeotroped using toluene (250 mL) to give an off white solid which was triturated using diethyl ether (50 mL) to give 3-(2,3-dihydrobenzofuran-5-yl)propanoic acid as a white solid (6.5 g, 80%). .sup.1H NMR (400 MHz, CDCl.sub.3) =7.04 (s, 1H), 6.93 (d, J=8.4, 1H), 6.7 (d, J=8.4 Hz, 1H), 4.55 (t, J=8.4 Hz, 2H), 3.18 (t, J=8.4 Hz, 2H), 2.89 (t, J=7.6 Hz, 2H), 2.64 (t, J=7.6 Hz, 2H).

[0187] A solution of 3-(2,3-dihydrobenzofuran-5-yl)propanoic acid (6.0 g, 31 mmol) in thionyl chloride (8 mL) was heated at 80 C. for 1 h. On completion of the reaction the thionyl chloride was removed in vacuo and the crude 3-(2,3-dihydrobenzofuran-5-yl)propanoyl chloride dissolved in anhydrous 1,2-dichloroethane (30 mL). In a separate flask aluminium trichloride (2 g, 15 mmol) was added to anhydrous 1,2-dichloroethane (40 mL) at 0 C. followed by the acid chloride solution (10 mL) drop-wise over 5 min and the resulting solution was stirred for 30 min at 0 C. A further portion of aluminium trichloride (3 g, 22.5 mmol) was added followed by drop-wise addition of the remaining acid chloride solution (20 mL) at 0 C. The reaction mixture was stirred at room temperature for 1 h or until completion, diluted with water and extracted using EtOAc (250 mL). The combined organic extracts were washed with 1N HCl (50 mL), 1N NaOH (50 mL), water (25 mL), brine (25 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 10% EtOAc-hexanes eluent to give 2,3,5,6-tetrahydro-7H-indeno[5,6-b]furan-7-one as a white solid (3.8 g, 70%). .sup.1H NMR (300 MHz, CD.sub.3OD) =7.36 (s, 1H), 6.91 (s, 1H), 4.61 (t, J=8.6 Hz, 3H), 3.26 (t, J=8.6 Hz, 2H), 3.05 (t, J=5.5 Hz, 3H), 2.68 (t, J=5.5 Hz, 2H).

[0188] 2,3,5,6-tetrahydro-7H-indeno[5,6-b]furan-7-one (1.5 g, 8.61 mmol) was dissolved in c.H.sub.2SO.sub.4 (6.0 mL) at 0 C. followed by drop-wise addition of f.HNO.sub.3:c.H.sub.2SO.sub.4, 1:1 (1.2 mL) stirring was continued at 0 C. for 1 h. The reaction mixture was added to ice-cold water (60 mL) and stirred for 10 min, the resulting light brown ppt was removed by filtration, washed with ice cold water (20 mL) and dried in vacuo to give 8-nitro-2,3,5,6-tetrahydro-7H-indeno[5,6-b]furan-7-one (1.2 g, 64%). .sup.1H NMR (300 MHz, CD.sub.3OD) =7.54 (s, 1H), 4.80 (t, J=8.6 Hz, 2H), 3.42 (t, J=8.6 Hz, 2H), 3.09 (t, J=5.6 Hz, 2H), 2.74 (t, J=5.6 Hz, 2H).

[0189] A solution of 8-nitro-2,3,5,6-tetrahydro-7H-indeno[5,6-b]furan-7-one (1.0 g, 4.57 mmol) in methanol (20 mL) 0 C. was treated with methane sulfonic acid (0.2 mL) followed by 20% palladium hydroxide (0.5 g). The reaction mixture was stirred under an atmosphere or hydrogen gas at 60 psi until completion. The reaction mixture was filtered through a bed of celite washing through with methanol (50 mL) and concentrated in vacuo. The residue was diluted with ethyl acetate (50 mL) and washed using sat. aq. NaHCO.sub.3 (50 mL), water (20 mL), brine (20 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 10% EtOAc-hexanes eluent to give 3,5,6,7-tetrahydro-2H-indeno[5,6-b]furan-8-amine as a white solid (0.5 g, 63%). .sup.1H NMR (300 MHz, CDCl.sub.3) =6.54 (s, 1H), 5.30 (s, 2H), 4.61 (t, J=8.7 Hz, 2H), 3.21 (t, J=8.7 Hz, 2H), 2.95 (t, J=5.5 Hz, 2H), 2.66 (t, J=5.5 Hz, 2H).

Intermediate I-17: Benzo[1,2-b:4,5-b]difuran-4-amine

[0190] ##STR00070##

[0191] Synthesis of 2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b]difuran-4-carboxylic acid was carried out from hydroquinone using procedures detailed by Monte et al., J. Med. Chem., 1996, 39, 2953-2961 to give the 2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b]difuran-4-carbaldehyde as a bright yellow solid; .sup.1H NMR (400 MHz, CDCl.sub.3): =10.27 (s, 1H), 6.87 (s, 1H), 4.67 (t, J=8.8 Hz, 2H), 4.59 (t, J=8.8 Hz, 2H), 4.59 (t, J=8.8 Hz, 2H), 3.46 (t, J=8.8 Hz, 2H), 3.18 (t, J=8.8 Hz, 2H).

[0192] The aldehyde (0.68 g, 3.58 mmol) was oxidized using silver (I) oxide (1.5 eq.) in 5% aqueous sodium hydroxide at rt for 20 days. The crude reaction mixture was filtered through celite, extracted using diethyl ether (250 mL) to remove unreacted aldehyde then the aqueous phase was acidified to pH 1 using 3.0 M aqueous HCl drop-wise at 0 C. The product was extracted using dichloromethane (250 mL) and the combined organics washed using brine (50 mL), dried (MgSO.sub.4) and concentrated in vacuo to give 2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b]difuran-4-carboxylic acid as a white solid (0.44 g; 60%).

[0193] Alternatively, the aldehyde (0.5 g, 2.77 mmol) in acetone (5.0 mL) was treated with sulfamic acid (0.4 g, 4.17 mmol) in two portions at 0 C. After 2 min a solution of sodium chlorite (0.32 g, 3.6 mmol) in water (1.0 mL) was added drop-wise and stirring continued at 0 C. for 4 h. The reaction mixture was diluted with water (20 mL) and extracted using 10% IPA/chloroform (220 mL). The combined organics were washed with water (25 mL), brine (25 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude solid was triturated with diethyl ether to give 2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b]difuran-4-carboxylic acid (0.4 g; 70%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): =6.86 (s, 1H), 4.52 (t, J=8.8 Hz, 2H), 4.47 (t, J=8.8 Hz, 2H), 3.30 (t, J=8.8 Hz, 2H), 3.10 (t, J=8.8 Hz, 2H). .sup.13C (100 MHz, DMSO-d.sub.6): =166.4, 154.2, 153.9, 128.9, 127.2, 111.4, 110.43, 71.9, 71.6, 31.5, 29.5.

[0194] 2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b]difuran-4-carboxylic acid (0.8 g, 3.88 mmol), 2, 3-dichloro-5,6-dicyanobenzoquinone (2.64 g, 11.65 mmol) in anhydrous dioxane (20 mL) was heated in a sealed tube at 120 C. for 18 h. The reaction mixture was cooled to room temperature and sat. aq. Na.sub.2S.sub.2O.sub.3 (30 mL) added before extraction with ethyl acetate (225 mL). The combined organics dried (Na.sub.2SO.sub.4) and concentrated in vacuo to give the crude benzo[1,2-b:4,5-b]difuran-4-carboxylic acid (1.5 g). The crude acid (1.5 g), triethylamine (2.05 mL) and diphenylphosphoryl azide (4.08 g, 14.85 mmol) in tertiary butanol (20 mL) was heated in a sealed tube at 90 C. for 12 h. The solution was cooled to room temperature, diluted with water (50 mL) and extracted using EtOAc (250 mL). The combined organic extracts were washed with water (25 mL), brine (25 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 10% EtOAc-hexanes eluent to give tert-butyl benzo[1,2-b:4,5-b]difuran-4-ylcarbamate (0.75 g) with minor impurities from the phosphine reagent, the product was dissolved in DCM (10 mL) and TFA (3.0 mL) added drop-wise over 5 min at 0 C. The reaction was stirred at ambient temperature for 2 h then added carefully to sat. aq. NaHCO.sub.3 (50 mL). The aqueous phase was extracted using DCM (230 mL) and the combined organic extracts were washed with water (25 mL), brine (25 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 10% EtOAc-hexanes eluent to give benzo[1,2-b:4,5-b]difuran-4-amine as an off-white solid (0.2 g, 30% over three steps). .sup.1H NMR (400 MHz, CDCl.sub.3): =7.6 (d, J=2.2 Hz, 1H), 7.53 (d, J=2.2 Hz, 1H), 7.12 (s, 1H), 6.78 (m, 2H), 4.17 (br.s., 1H).

Intermediate I-23: 4-(2-Aminoethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)benzenesulfonamide

Step A: tert-Butyl (4-sulfamoylphenethyl)carbamate

[0195] ##STR00071##

[0196] To a solution of 4-(2-aminoethyl)benzenesulfonamide (5 g, 24.97 mmol) in THF (100 mL) was added Boc.sub.2O (6.5 g, 29.96 mmol, 6.88 mL) and triethylamine (5-18, 49.94 mmol, 6.95 mL). The mixture was stirred at 25 C. for 2 hours before the reaction was quenched with water (100 mL) and extracted into dichloromethane (3150 mL). The combined organic extracts were washed with brine (2100 mL), dried (Na.sub.2SO.sub.4), filtered and concentrated in vacuo. The crude product was triturated with ethyl acetate (100 mL) and then isolated by filtration to afford the title compound as yellow solid (7.0 g, 93% yield). The crude product was used in next step directly without further purification.

[0197] .sup.1H NMR (DMSO-d.sub.6) 7.75-7.73 (d, 2H), 7.39-7.37 (d, 2H), 7.30 (s, 2H), 6.93 (s, 1 H), 3.17-3.14 (m, 2H), 2.79-2.75 (m, 2H) and 1.37 (s, 9H).

[0198] LCMS: m/z 323.0 (M+Na).sup.+

Step B: tert-Butyl 4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-phenethylcarbamate

[0199] ##STR00072##

[0200] To a mixture of tert-butyl (4-sulfamoylphenethyl)carbamate (3.0 g, 10.0 mmol) in THF (30 mL) was added sodium methoxide (539.6 mg, 10.0 mmol). The mixture was stirred at 25 C. for 2 hours and then 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (3.0 g, 12.0 mmol) in THF (10 mL) was added drop-wise. The mixture was stirred at 70 C. for another 0.5 hours before the reaction was quenched with water (30 mL) and then adjusted pH to 5-6 with hydrochloric acid (1 M). The resulting solid was filtered and dried in vacuo to afford the title compound as white solid (4.2 g, 84% yield). The crude product was used in next step directly without further purification.

[0201] .sup.1H NMR (DMSO-d.sub.6) 10.64 (s, 1H) 8.08 (s, 1H), 7.86-7.84 (d, 2H), 7.44-7.42 (d, 2H), 6.93 (s, 2H), 3.18-3.17 (m, 2H), 2.81-2.75 (m, 7H), 2.54-2.52 (m, 3H), 1.98-1.88 (m, 4H) and 1.35 (s, 9H)

[0202] LCMS: m/z 444.1 (M+H56).sup.+

Step C: 4-(2-Aminoethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)-carbamoyl)-benzenesulfonamide

[0203] ##STR00073##

[0204] To a solution of tert-butyl 4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethylcarbamate (4.2 g, 8.4 mmol) in dichloromethane (25 mL) was added a solution of hydrogen chloride in ethyl acetate (25 mL, 100.0 mmol). The reaction was allowed to stir at 25 C. for 2 hours before the mixture was poured into water (50 mL) and then adjusted to pH 7 with aqueous sodium hydroxide solution (1 M). The resulting precipitate was filtered and washed with methanol (20 mL) to afford the title compound, Intermediate 1-23 (2.8 g, 75% yield) as a white solid. 1H NMR (DMSO-d.sub.6) 10.97 (s, 1H) 8.55 (s, 1H), 7.92 (s, 2H) 7.82-7.80 (d, 2H), 7.44-7.42 (d, 2H), 6.83 (s, 2H), 2.98-2.91 (m, 2H), 2.89-2.70 (m, 2H), 2.68-2.63 (m, 5H), 2.61-2.59 (m, 2H) and 1.85-1.81 (m, 4H).

[0205] LCMS m/z 400.1 (M+H).sup.+

Sulfonylurea Synthesis

Example 1: 4-Acetyl-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl) benzenesulfonamide

[0206] ##STR00074##

4-Acetylbenzenesulfonamide (100 mg, 0.50 mmol) was treated as per general method A to afford the titled compound as a white solid (31 mg, 16%): .sup.1H NMR (600 MHz, DMSO-d.sub.6) 11.03 (br s, 1H), 8.08 (d, J=8.5 Hz, 2H), 7.99 (d, J=8.5 Hz, 2H), 7.07 (br s, 1H), 6.87 (s, 1H), 2.75 (t, J=7.4 Hz, 4H), 2.62 (s, 3H), 2.56 (t, J=7.4 Hz, 4H), 1.90 (p, J=7.4 Hz, 4H); .sup.13C NMR (151 MHz, DMSO-d.sub.6) 197.3, 151.8, 142.6, 138.9, 136.9, 129.7, 128.2, 127.2, 117.1, 32.4, 30.1, 26.9, 24.9; HRMS (ESI-TOF) m/z calcd for C.sub.21H.sub.21N.sub.2O.sub.4S [MH].sup. 397.1228, found 397.1225; LC-MS m/z 399.1 [M+H].sup.+, purity >95% (ELSD).

Example 2: 5-Chloro-N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl) sulfamoyl)phenethyl)-2-methoxybenzamide

[0207] ##STR00075##

[0208] 5-Chloro-2-methoxy-N-(4-sulfamoylphenethyl) benzamide (300 mg, 0.81 mmol) was treated as per general method A to afford the titled compound as a white solid (325 mg, 70%); .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.74 (br s, 1H), 8.27 (t, J=5.7 Hz, 1H), 8.01 (s, 1H), 7.86 (d, J=8.3 Hz, 2H), 7.65 (d, J=2.8 Hz, 1H), 7.50 (dd, J=8.9, 2.8 Hz, 1H), 7.47 (d, J=8.3 Hz, 2H), 7.13 (d, J=8.9 Hz, 1H), 6.89 (s, 1H), 3.78 (s, 3H), 3.59-3.49 (m, 2H), 2.92 (t, J=7.1 Hz, 2H), 2.75 (t, J=7.4 Hz, 4H), 2.53 (t, J=7.4 Hz, 4H), 1.89 (p, J=7.5 Hz, 4H); .sup.13C NMR (151 MHz, DMSO-d.sub.6) 163.5, 155.6, 153.8, 144.4, 142.8, 136.9, 136.9, 131.4, 129.4, 129.4, 128.9, 127.1, 124.6, 124.2, 117.4, 114.0, 56.1, 40.2, 34.6, 32.3, 30.0, 24.9; HRMS (ESI-TOF) m/z calcd for C.sub.29H.sub.29ClN.sub.3O.sub.5S [MH].sup. 566.1522, found 566.1543; LC-MS m/z 566.2 [MH].sup., purity >95% (ELSD).

Example 3: N-(4-(N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl) phenethyl)-5-methylpyrazine-2-carboxamide

[0209] ##STR00076##

[0210] 5-Methyl-N-(4-sulfamoylphenethyl)pyrazine-2-carboxamide (Intermediate I-3) (100 mg, 0.312 mmol) was treated as per general method A to afford a crude white solid (72 mg). Further purification by HPLC yielded a white solid (14 mg, 9%). .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.79 (br s, 1H), 9.03 (d, J=1.4 Hz, 1H), 8.96 (t, J=6.0 Hz, 1H), 8.59 (m, 1H), 7.79 (d, J=8.3 Hz, 2H), 7.47 (d, J=8.3 Hz, 2H), 7.30 (s, 1H), 6.86 (s, 1H), 3.58-3.52 (m, 2H), 2.94 (t, J=7.3 Hz, 2H), 2.75 (t, J=7.5 Hz, 4H), 2.58 (br s, 3H), 2.54 (t, J=7.5 Hz, 4H), 1.88 (p, J=7.5 Hz, 4H); .sup.13C NMR (151 MHz, DMSO-d.sub.6) 162.8, 156.7, 152.1, 143.4, 142.7, 142.6, 142.3, 141.9, 136.8, 129.9, 129.0, 128.5, 126.9, 125.6, 117.0, 40.0, 34.6, 32.4, 30.1, 24.9, 21.2; HRMS (ESI-TOF) m/z calcd for C.sub.27H.sub.30N.sub.5O.sub.4S [M+H].sup.+ 520.2020, found 520.2030; LC-MS m/z 518.2 [MH].sup., purity >95% (ELSD).

Example 4: N-(4-(N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl) phenethyl)-5-methylisoxazole-3-carboxamide

[0211] ##STR00077##

[0212] 5-Methyl-N-(4-sulfamoylphenethyl) isoxazole-3-carboxamide (Intermediate I-4) (14 mg, 0.044 mmol) was treated as per general method A. Further MPLC purification using 0.05% formic acid.sub.(aq)/0.05% formic acid in CH.sub.3CN gradient elution. Lyophilisation of the appropriate fractions afforded the titled compound as a white solid (14 mg, 62%): .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.73 (br s, 1H), 8.81 (t, J=5.8 Hz, 1H) 8.09 (s, 1H), 7.86 (d, J=8.2 Hz, 2H), 7.47 (d, J=8.2 Hz, 2H), 6.92 (s, 1H), 6.49 (s, 1H), 3.49 (m, 2H), 2.93 (t, J=7.2 Hz, 2H), 2.76 (t, J=7.2 Hz, 4H), 2.52 (m, 4H), 2.44 (s, 3H), 1.90 (p, J=7.2 Hz, 4H); .sup.13C NMR (151 MHz, DMSO) 171.0, 158.7, 158.5, 148.9, 145.0, 142.9, 137.8, 137.1, 129.1, 128.5, 127.2, 117.9, 101.1, 39.7, 34.4, 32.3, 29.9, 24.9, 11.70; HRMS (ESI-TOF) m/z calcd for C.sub.26H.sub.27N.sub.4O.sub.5S [MH].sup. 507.1708, found 507.1709; LC-MS m/z 509.4 [M+H].sup.+, purity >95% (ELSD).

Example 5: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-(7-methoxy-4,4-dimethyl-1,3-dioxo-3,4-dihydroisoquinolin-2(1H)-yl)ethyl)benzenesulfonamide

[0213] ##STR00078##

[0214] 4-(2-(7-Methoxy-4,4-dimethyl-1,3-dioxo-3,4-dihydroisoquinolin-2(1H)-yl)ethyl)benzene sulfonamide (Intermediate I-5) (111 mg, 0.276 mmol) was treated as per general method A to afford the titled compound as a white solid (85 mg, 52%): .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.72 (br s, 1H), 7.91 (s, 1H), 7.80 (d, J=8.0 Hz, 2H), 7.58 (d, J=8.7 Hz, 1H), 7.53 (d, J=2.9 Hz, 1H), 7.39 (d, J=8.0 Hz, 2H), 7.29 (dd, J=8.7, 2.9 Hz, 1H), 6.88 (s, 1H), 4.13 (t, J=7.5 Hz, 2H), 3.83 (s, 3H), 2.93 (t, J=7.4 Hz, 2H), 2.76 (t, J=7.4 Hz, 4H), 2.55 (t, J=7.4 Hz, 4H), 1.90 (p, J=7.4 Hz, 4H), 1.42 (s, 6H); .sup.13C NMR (151 MHz, DMSO) 176.3, 163.1, 158.0, 150.1, 143.5, 142.8, 137.1, 136.9, 129.1, 129.0, 127.4, 127.1, 125.6, 124.1, 121.4, 117.4, 110.4, 55.3, 42.3, 40.3, 33.0, 32.3, 30.0, 28.8, 24.9; HRMS (ESI-TOF) m/z calcd for C.sub.33H.sub.34N.sub.3O.sub.6S [MH].sup. 600.2174, found 600.2183; LC-MS m/z 602.4 [M+H].sup.+, purity >95% (ELSD).

Example 6: 3-Ethyl-N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl) sulfamoyl)phenethyl)-4-methyl-2-oxo-2,5-dihydro-1H-pyrrole-1-carboxamide

[0215] ##STR00079##

[0216] 3-Ethyl-4-methyl-2-oxo-N-(4-sulfamoylphenethyl)-2,5-dihydro-1H-pyrrole-1-carboxamide (100 mg, 0.29 mmol) was treated as per general method A to afford the titled compound as a white solid (78 mg, 50%). .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.73 (br s, 1H), 8.38 (t, J=5.8 Hz, 1H), 8.04 (s, 1H), 7.84 (d, J=8.4 Hz, 2H,), 7.45 (d, J=8.4 Hz, 2H), 6.90 (s, 1H), 4.16 (s, 2H), 3.52-3.45 (m, 2H), 2.89 (t, J=7.2 Hz, 2H), 2.75 (t, J=7.5 Hz, 4H), 2.52 (d, J=7.4 Hz, 4H), 2.18 (q, J=7.5 Hz, 2H,), 2.00 (s, 3H), 1.90 (p, J=7.5 Hz, 4H), 0.97 (t, J=7.5 Hz, 3H). .sup.13C NMR (151 MHz, DMSO-d.sub.6) 171.8, 152.1, 151.7, 149.8, 144.7, 143.0, 138.6, 137.2, 131.9, 129.1, 129.0, 127.4, 117.8, 51.9, 40.2, 35.3, 32.5, 30.1, 25.0, 16.0, 12.9, 12.8. HRMS (ESI-TOF) m/z: calcd for C.sub.29H.sub.33N.sub.4O.sub.5S [MH].sup. 549.2177; found 549.2169. LCMS: m/z 573.47 [M+Na].sup.+, purity >95% (ELSD).

Example 7: Ethyl 5-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-furan-3-carboxylate

[0217] ##STR00080##

[0218] 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A2) and ethyl 5-sulfamoylfuran-3-carboxylate were used in general method B1. The reaction mixture was quenched using water (50 mL), extracted using ethyl acetate (225 mL) and the organics washed with brine (25 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 50% EtOAc-hexanes eluent to give the titled compound as a white solid (0.45 g, 63%). .sup.1H NMR (300 MHz, DMSO-d.sub.6) =8.31 (s, 1H), 7.59 (s, 1H), 6.77 (s, 1H), 4.22 (q, J=7.2 Hz, 2H), 2.75 (t, J=7.3 Hz, 4H), 2.65 (t, J=7.3 Hz, 4H) 1.90 (pent, J=7.6 Hz, 4H), 1.26 (t, J=7.2 Hz, 3H).

Example 8: 5-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)furan-3-carboxylic acid

[0219] ##STR00081##

[0220] Ethyl 5-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)furan-3-carboxylate (Example 7) (0.1 g, 0.24 mmol) in THF (8 mL) at 0 C. was treated with a solution of LiOH (0.1 g, 2.4 mmol) in water (2 mL). The cooling bath was removed and the reaction mixture stirred for 3 h. The solution was acidified using 10% citric acid and immediately extracted using ethyl acetate (225 mL). The organics were washed using water (20 mL), brine (20 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by reversed phase HPLC to give the titled compound as a white solid (5.0 mg, 5%). .sup.1H NMR (400 MHz, CD.sub.3OD): =8.14 (s, 1H), 7.28 (s, 1H), 6.93 (s, 1H), 2.85 (t, J=7.6 Hz, 4H), 2.74 (t, J=7.6 Hz, 4H), 2.04 (quin, J=7.6 Hz, 4H).

Example 9: Methyl 3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)benzoate

[0221] ##STR00082##

[0222] 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A1) was added directly to methyl 3-sulfamoylbenzoate (Intermediate I-9) (0.447 g, 2.07 mmol, 1.20 equiv) at ambient temperature and the mixture was stirred overnight. The reaction mixture was poured into chilled water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The residue obtained was purified by column chromatography on silica gel using 0-10% gradient of methanol in dichloromethane to give methyl 3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)benzoate as a light-brown solid (0.36 g, 50%).

Example 10: 3-(N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-benzoic acid

[0223] ##STR00083##

[0224] Methyl 3-(N-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)sulfamoyl)benzoate (Example 9) (0.25 g, 0.603 mmol) was dissolved in a mixture of tetrahydrofuran:methanol:water (9 mL, 1:1:1) and the mixture was cooled to 0 C. Lithium hydroxide monohydrate (0.75 g, 1.81 mmol, 3 eq) was added and the mixture stirred at ambient temperature for 3 h. Upon completion, the reaction mixture was poured into chilled water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The product was purified by reverse phase preparative HPLC to afford the titled compound as a white solid (0.017 g, 3%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): =13.26 (bs, 1H), 8.43 (s, 1H), 8.13-8.08 (m, 2H), 7.99 (bs, 1H), 7.67 (t, J=8.0 Hz, 1H), 6.87 (s, 1H), 6.52 (s, 1H), 2.75 (t, J=7.2 Hz, 4H), 2.55 (t, J=7.6 Hz, 4H), 1.89 (quin, J=7.6 Hz, 4H). LCMS, Purity: 96%, m/z 400.98 (M+H.sup.+). HRMS (FAB.sup.+) calcd for C.sub.20H.sub.20N.sub.2O.sub.5S [M+H].sup.+: 401.1093, found: 401.4514.

Example 11: 3-(N-(1,2,3,5,6,7-Hexahydro-s-indacen-4-ylcarbamoyl)sulfamoyl)-benzamide

[0225] ##STR00084##

[0226] 3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)benzoic acid (Example 10) (0.06 g, 0.074 mmol) was dissolved in anhydrous N,N-dimethylformamide (4 mL) and the solution cooled to 0 C. Diisopropylethylamine (3.0 eq) and HATU (2.0 eq) were added and the mixture stirred at 0 C. for 15 min. Ammonium chloride (3.0 eq) was added and the mixture stirred at ambient temperature for 5 h. Upon completion the reaction mixture was poured into brine (20 mL) and extracted with ethyl acetate (210 mL). The combined organic extracts were washed with brine (10 mL), dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude residue was purified by reverse phase preparative HPLC to afford the titled compound as a white solid (0.011 g, 37%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): =8.23 (d, J=9.2 Hz, 2H), 8.02 (s, 1H), 7.89 (d, J=7.6 Hz, 1H), 7.84 (d, J=7.6 Hz, 1H), 7.42 (t, J=8.0 Hz, 1H), 7.38 (s, 1H), 7.33 (s, 1H). 6.74 (s, 1H), 2.73 (t, J=6.8 Hz, 4H), 2.62 (t, J=6.8 Hz, 4H), 1.87 (quin, J=7.6 Hz, 4H). LCMS, Purity: 93%, m/z 400.05 (M+H.sup.+). HRMS (FAB.sup.+) calcd for C.sub.20H.sub.21N.sub.3O.sub.4S [M+H].sup.+: 400.1253, found: 400.1378.

Example 12: N-(3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-phenyl)pent-4-ynamide

[0227] ##STR00085##

[0228] 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A2) and N-(3-sulfamoylphenyl)pent-4-ynamide (Intermediate 1-12) were used in general method B3 to give the titled compound as a white solid (116 mg, 61%). .sup.1H NMR (400 MHz, CD.sub.3OD) =8.18 (s, 1H), 7.81 (d, J=8.3 Hz, 1H), 7.68 (d, J=8.3 Hz, 1H), 7.43 (dd, J=8.3, 7.8 Hz, 1H), 6.87 (s, 1H), 2.79 (t, J=7.2 Hz, 4H), 2.67-2.60 (m, 4H), 2.60-2.48 (m, 4H), 2.28-2.22 (m, 1H), 2.04-1.89 (m, 4H). .sup.13C NMR (101 MHz, CD.sub.3OD) =170.9, 143.3, 143.0, 138.8, 137.7, 128.7, 128.3, 126.4, 122.8, 122.0, 117.9, 117.7, 82.0, 68.9, 35.4, 32.4, 29.9, 25.1, 13.9. HRMS (ESI) calcd. for C.sub.24H.sub.26N.sub.3O.sub.4S [M+H] 452.1639, found 452.1658.

Example 13: 3-(4-(N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-phenyl)-N-(prop-2-yn-1-yl)propanamide

[0229] ##STR00086##

[0230] 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A2) and N-(prop-2-yn-1-yl)-3-(4-sulfamoylphenyl)propanamide (Intermediate 1-13) were used in general method B3 to give the titled compound as a white solid (120 mg, 68%). .sup.1H NMR (400 MHz, CD.sub.3OD) =7.91 (d, J=7.8 Hz, 2H), 7.39 (d, J=7.8 Hz, 2H), 6.98 (s, 1H), 3.95 (d, J=2.9 Hz, 2H), 3.03 (t, J=7.7 Hz, 2H), 2.85 (t, J=7.4 Hz, 4H), 2.62 (t, J=6.9 Hz, 4H), 2.55-2.46 (m, 2H), 2.25 (t, J=2.6 Hz, 1H), 2.02 (m, 4H). .sup.13C NMR (101 MHz, CD.sub.3OD) =172.0, 147.2, 144.1, 143.8, 137.5, 129.0, 128.8, 128.1, 127.4, 126.5, 118.9, 79.2, 71.0, 36.8, 32.8, 32.8, 31.2, 30.7, 28.8, 28.7, 25.4, 25.3. HRMS (ESI) calcd. for C.sub.25H.sub.28N.sub.3O.sub.4S [M+H] 466.1795, found 466.1794.

Example 14: N-(4-(N-(1,2,3,5,6,7-Hexahydro-s-indacen-4-ylcarbamoyl)sulfamoyl)-phenethyl)-2-(methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)acetamide

[0231] ##STR00087##

[0232] 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A1) and 2-(methyl(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)-N-(4-sulfamoylphenethyl)acetamide (Intermediate 1-14) were used in general method B2 to give the titled compound as an orange solid (0.003 g, 1%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): =10.74 (s, 1H), 8.51 (d, J=8.8 Hz, 1H), 8.31 (t, J=7.6 Hz, 1H), 8.09-7.96 (m, 1H), 7.82 (d, J=8.4 Hz, 2H), 7.41 (d, J=7.6 Hz, 2H), 6.89 (s, 1H), 6.42-6.32 (m, 1H), 4.74 (bs, 2H), 3.44-3.30 (m, 5H), 2.80 (t, J=7.6 Hz, 2H), 2.73-2.69 (m, 4H), 2.61-2.50 (m, 4H), 1.92-1.88 (m, 4H). LCMS, Purity: 92.20%, m/z 632.35 (MH.sup.+).

Example 15: 2-(7-(Dimethylamino)-2-oxo-2H-chromen-4-yl)-N-(4-(N-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)sulfamoyl)phenethyl)acetamide

[0233] ##STR00088##

[0234] 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A1) and 2-(7-(dimethylamino)-2-oxo-2H-chromen-4-yl)-N-(4-sulfamoylphenethyl)acetamide (Intermediate 1-15) were used in general method B2 to give the titled compound as a pale-yellow solid (0.008 g, 0.44%). .sup.1H NMR (400 MHz, DMSO-d.sub.6): =9.53 (s, 1H), 8.29 (t, J=4.8 Hz, 1H), 8.22 (s, 1H), 7.68 (d, J=8.0 Hz, 2H), 7.46 (d, J=8.8 Hz, 1H), 8.42 (s, 1H), 7.16 (d, J=8.0 Hz, 2H), 6.74-6.70 (m, 2H), 6.54 (d J=2.4 Hz, 1H), 5.99 (s, 1H), 3.56-3.52 (m, 2H), 3.48 (t, J=6.0 Hz, 2H), 3.31-3.24 (m, 2H), 2.76-2.70 (m, 4H), 3.02 (s, 6H), 2.63 (t, J=7.2 Hz, 4H), 1.88 (quin, J=7.6 Hz, 4H). LCMS, Purity: 92.26%, m/z 629.40 (MH.sup.+).

Example 16: Ethyl 5-(N-((3,5,6,7-tetrahydro-2H-indeno[5,6-b]furan-8-yl)carbamoyl)-sulfamoyl)furan-3-carboxylate

[0235] ##STR00089##

[0236] 8-Isocyanato-3,5,6,7-tetrahydro-2H-indeno[5,6-b]furan (prepared using general method A1 from Intermediate 1-16) and ethyl 5-sulfamoylfuran-3-carboxylate were used in general method B1 to give ethyl 5-(N-((3,5,6,7-tetrahydro-2H-indeno[5,6-b]furan-8-yl)carbamoyl)sulfamoyl)furan-3-carboxylate as a pale-brown solid (0.25 g, 50%). .sup.1H NMR (300 MHz, DMSO-d.sub.6) 8.32 (s, 1H), 7.17 (s, 1H), 6.77 (d, J=5.2 Hz, 1H), 4.43 (t, J=8.6 Hz, 2H), 4.23 (q, J=7.1 Hz, 2H), 3.07 (t, J=8.6 Hz, 2H), 2.71 (t, J=7.3 Hz, 2H), 2.63 (t, J=7.3 Hz, 2H), 1.89 (p, J=7.4 Hz, 2H), 1.26 (t, J=7.1 Hz, 3H).

Example 17: Ethyl 5-(N-(benzo[1,2-b:4,5-b]difuran-4-ylcarbamoyl)sulfamoyl)furan-3-carboxylate

[0237] ##STR00090##

[0238] 4-isocyanatobenzo[1,2-b:4,5-b]difuran (prepared using general method A1 from Intermediate 1-17) and ethyl 5-sulfamoylfuran-3-carboxylate were used in general method B1 to give ethyl 5-(N-(benzo[1,2-b:4,5-b]difuran-4-ylcarbamoyl)sulfamoyl)furan-3-carboxylate as a white solid (0.05 g, 53%). .sup.1H NMR (300 MHz, CD.sub.3OD) =8.25 (s, 1H), 7.72 (d, J 2.1 Hz, 1H), 7.63 (d, J 2.1 Hz, 1H), 7.46 (s, 1H), 7.27 (s, 1H), 6.93 (s, 1H), 6.89 (d, J 2.1 Hz, 1H), 6.86 (d, J 2.1 Hz, 1H), 4.30 (q, J 6.9 Hz, 2H), 1.4 (t, J 6.9 Hz, 3H).

Example 18: Methyl 2-(N-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)sulfamoyl)-isonicotinate

[0239] ##STR00091##

[0240] To a solution of 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (0.20 g, 1.15 mmol) in anhydrous THF (5 mL), triethylamine (0.35 g, 3.47 mmol, 3.0 eq) was added followed by triphosgene (0.265 g, 0.86 mmol, 0.5 eq) at 0 C. and the mixture was stirred at ambient temperature for 3 h. The mixture was cooled to 0 C., methyl 2-sulfamoylisonicotinate (0.27 g, 1.27 mmol, 1.1 eq) added and stirring continued at ambient temperature overnight. Upon completion the reaction mixture was poured into brine and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The crude product was purified by column chromatography on silica gel using 20-50% gradient of EtOAc-hexanes eluent to give methyl 2-(N-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)sulfamoyl)isonicotinate as a light brown solid (0.31 g, 65%).

Example 19: 3-(1-(3-Aminopropyl)-1H-1,2,3-triazol-4-yl)-N-(3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)phenyl)propanamide

[0241] ##STR00092##

[0242] N-(3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)phenyl)pent-4-ynamide (Example 12) and 3-azidopropan-1-amine were used in general method C to give the titled compound as a white solid (6 mg, 43%). .sup.1H NMR (600 MHz, CD.sub.3OD) =7.85 (s, 1H), 7.55 (t, J=3.8 Hz, 2H), 7.50 (d, J=8.0 Hz, 2H), 7.29 (t, J=7.9 Hz, 1H), 6.78 (s, 1H), 4.26 (t, J=6.4 Hz, 2H), 3.00 (t, J=6.6 Hz, 2H), 2.71 (t, J=7.3 Hz, 4H), 2.64-2.50 (m, 8H), 1.94-2.02 (m, 2H), 1.92-1.83 (m, 4H). .sup.13C NMR (151 MHz, CD.sub.3OD) =173.0, 147.4, 146.8, 144.7, 144.6, 139.5, 139.2, 131.6, 130.0, 129.8, 124.2, 123.9, 123.2, 119.5, 118.6, 48.3, 37.7, 34.0, 31.6, 26.7, 26.6, 22.9. HRMS (ESI) calcd. for C.sub.27H.sub.34N.sub.7O.sub.4S [M+H] 552.2387, found 552.2368.

Example 20: N-(3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-phenyl)-3-(1-(3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)propyl)-1H-1,2,3-triazol-4-yl)propanamide

[0243] ##STR00093##

[0244] N-(2-Azidopropyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine was synthesized by the methods contained in Chun Li, Etienne Henry, Naresh Kumar Mani, Jie Tang, Jean-Claude Brochon, Eric Deprez, and Juan Xie, Eur. J. Org. Chem., 2010, 2395-2405. To a solution of 4-chloro-7-nitrobenzo[c][1,2,5]oxadiazole (300 mg, 1.5 mmol) in THF (10 mL) was added 3-azidopropyl amine (160 mg, 1.65 mmol) and Cs.sub.2CO.sub.3 (480 mg, 1.5 mmol). The reaction mixture was stirred at 50 C. for 4 h. The reaction mixture was partitioned between EtOAc (50 mL) concentrated in vacuo. The residue was purified by column chromatography on silica gel using 30% EtOAc-petroleum ether eluent to afford N-(2-Azidopropyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine (240 mg, 76%). .sup.1H NMR (400 MHz, CDCl.sub.3): =8.50 (d, J=8.8 Hz, 1H), 6.57 (s, 1H, NH), 6.23 (d, J=8.8 Hz, 1H), 3.66 (q, J=6.8 Hz, 2H), 3.59 (J=6.0 Hz, 2H), 2.00-2.16 (m, 2H). .sup.13C NMR (101 MHz, CDCl.sub.3) 144.2, 144.0, 143.8, 136.7, 123.7, 98.8, 49.1, 41.6, 27.6. HRMS (ESI): calcd. for C.sub.9H.sub.10N.sub.7O.sub.3 264.0840; found 264.0711.

[0245] N-(3-(N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)phenyl)pent-4-ynamide (Example 12) (10 mg, 0.022 mmol) and N-(2-azidopropyl)-7-nitrobenzo[c][1,2,5]oxadiazol-4-amine (7.0 mg, 0.026 mmol), 10 mol % THPTA, 5 mol % CuSO.sub.4, 10 mol % sodium ascorbate in DMSO (500 uL) were stirred at room temperature for 12 h. The reaction mixture was subjected to purification using reverse phase (Reveleris flash column chromatography, 4 g, 18 mL/min., mobile phase; 10 mmol aqu. NH.sub.4CO.sub.3, MeCN) and freeze dried to give the product as a white solid (7.0 mg, 44%). .sup.1H NMR (600 MHz, CD.sub.3OD) =8.46 (d, J=8.7 Hz, 1H), 8.18 (s, 1H), 7.79 (d, J=8.4 Hz, 1H), 7.67 (d, J=7.9 Hz, 1H), 7.61 (s, 1H), 6.94 (s, 1H), 6.15 (d, J=9.0 Hz, 1H), 4.46 (t, J=6.7 Hz, 2H), 3.09 (t, J=7.0 Hz, 2H), 2.82 (t, J=7.4 Hz, 4H), 2.77 (t, J=7.0 Hz, 2H), 2.70-2.56 (m, 6H), 2.37-2.26 (m, 2H), 1.99 (q, J=7.3 Hz, 4H). .sup.13C NMR (151 MHz, CD.sub.3OD) =172.9, 147.9, 145.4, 140.5, 139.0, 138.4, 130.6, 129.1, 128.0, 125.6, 124.2, 123.6, 120.3, 119.6, 112.4, 70.6, 48.9, 37.2, 34.3, 34.2, 31.7, 30.2, 26.8, 22.3; HRMS (ESI) calcd. for C.sub.33H.sub.34N.sub.10O.sub.7S [MH] 713.2260, found 713.2290.

Example 21: N-(3-(4-(3-((3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)phenyl) amino)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)propyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide

[0246] ##STR00094##

[0247] To a solution of biotin (0.4 g, 1.63 mmol) and 3-azidopropylamine (0.2 g, 1.96 mmol) in dry DMF (10.0 ml) was added HBTU (0.93 g, 2.45 mmol) followed by DIPEA (428 uL, 2.45 mmol). The reaction mixture was stirred at RT for 12 h. The reaction was monitored by LCMS and after the completion of reaction, it was diluted with EtOAc (50 mL) washed with H.sub.2O (25 mL), brine (25 mL). The organic layer was separated; dried (MgSO.sub.4) and evaporated to give the crude product. The crude product was purified by column chromatography on silica gel using 50% EtOAc-Hexane eluent to isolate N-(3-azidopropyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide as a white solid (0.13 g, 24%). .sup.1H NMR (400 MHz, CD.sub.3OD) =4.52 (dd, J=7.9, 5.0 Hz, 1H), 4.32 (dd, J=7.9, 4.5 Hz, 1H), 3.36 (t, J=6.7 Hz, 2H), 3.28 (d, J=6.8 Hz, 2H), 3.21-3.14 (m, 1H), 2.93 (dd, J=12.8, 5.0 Hz, 1H), 2.75 (d, J=12.8 Hz, 1H), 2.20 (t, J=7.3 Hz, 2H), 1.78 (q, J=6.8 Hz, 2H), 1.74-1.57 (m, 4H), 1.45 (q, J=7.5 Hz, 2H). .sup.13C NMR (101 MHz, CD.sub.3OD) =173.5, 163.4, 61.0, 59.3, 54.7, 48.2, 39.4, 35.8, 34.8, 27.7, 27.5, 27.2, 24.6.

[0248] N-(3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)phenyl)pent-4-ynamide (Example 12) (1.0 mmol) and N-(3-azidopropyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide (2.0 mmol), 10 mol % THPTA, 5 mol % CuSO.sub.4, 10 mol % sodium ascorbate in DMSO were stirred at room temperature for 12 h. The reaction mixture was purified using reverse phase column chromatography to give the titled compound as a white solid (8.0 mg, 31%); .sup.1H NMR (600 MHz, CD.sub.3OD) =8.26 (s, 1H), 7.83-7.68 (m, 3H), 7.50-7.43 (m, 1H), 6.92 (s, 1H), 4.48 (dd, J=8.0, 4.8 Hz, 1H), 4.41-4.22 (m, 3H), 3.18 (dd, J=6.9, 3.5 Hz, 1H), 3.14 (td, J=6.7, 1.7 Hz, 2H), 3.12-3.06 (m, 2H), 2.90 (dd, J=12.8, 4.9 Hz, 1H), 2.81 (t, J=7.7 Hz, 4H), 2.77 (d, J=7.1 Hz, 1H), 2.71 (s, 1H), 2.62 (t, J=7.3 Hz, 4H), 2.19 (td, J=7.4, 1.7 Hz, 2H), 2.05-2.01 (m, 2H), 2.00-1.95 (m, 4H), 1.76-1.57 (m, 4H), 1.43 (q, J=7.6, 7.1 Hz, 2H). .sup.13C NMR (151 MHz, CD.sub.3OD) =174.8, 174.8, 171.6, 171.5, 164.5, 146.2, 143.6, 139.1, 137.7, 129.1, 129.1, 128.7, 128.1, 126.5, 123.7, 122.9, 122.4, 122.2, 120.9, 118.4, 118.4, 118.3, 118.2, 118.2, 117.2, 110.5, 69.0, 61.9, 60.2, 55.6, 39.8, 36.1, 36.0, 35.8, 35.4, 35.4, 32.6, 32.6, 30.0, 29.7, 29.7, 28.6, 28.3, 28.0, 25.3, 25.2, 25.2, 20.9. HRMS (ESI) calcd. for C.sub.37H.sub.48N.sub.9O.sub.6S.sub.2[M+H] 778.3163, found 778.3145.

Example 22: N-((1-(3-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)phenyl)-1H-1,2,3-triazol-4-yl)methyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide

[0249] ##STR00095##

[0250] 5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)-N-((1-(3-sulfamoylphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentanamide was synthesized using 5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)-N-(prop-2-yn-1-yl)pentanamide (1.0 mmol) and 3-azidobenzenesulfonamide (2.0 mmol), 10 mol % THPTA, 5 mol % CuSO.sub.4, 10 mol % NaAsc in DMSO were stirred at room temperature for 12 h. The formation of product was observed in LCMS. After completion of the reaction, the reaction mixture was subjected to HPLC purification (Reveleris flash column chromatography, 4 g, 18 mL/min., mobile phase; 10 mmol aq. NH.sub.4CO.sub.3, MeCN) to isolate 5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)-N-((1-(3-sulfamoylphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentanamid as a white solid (24 mg, 47%) which was used directly.

[0251] To a solution of 5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)-N-((1-(3-sulfamoylphenyl)-1H-1,2,3-triazol-4-yl)methyl)pentanamide (15 mg, 0.031 mmol) in THF (5.0 mL) under a nitrogen atmosphere was added DIPEA (605 L, 0.037 mmol). This mixture was stirred at room temperature for 15 min. A solution of 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (prepared using general method A2) (705 mg, 0.037 mmol) in THF was added drop-wise. The reaction mixture was stirred at room temperature overnight then the solvent was removed in vacuo to give crude compound which was purified by reversed phase column chromatography using 10 mM aq. (NH.sub.4).sub.2CO.sub.3 and MeCN mobile phase to isolate the titled compound as a white solid (5.2 mg, 24%). .sup.1H NMR (600 MHz, CD.sub.3OD) =8.53 (d, J=2.4 Hz, 1H), 8.50 (d, J=7.5 Hz, 1H), 8.22-8.11 (m, 2H), 7.86-7.78 (m, 1H), 6.99 (s, 1H), 4.62 (s, 2H), 4.57-4.49 (m, 1H), 4.38-4.31 (m, 1H), 3.27-3.20 (m, 1H), 3.00-2.84 (m, 4H), 2.78-2.70 (m, 4H), 2.36 (t, J=7.2 Hz, 2H), 2.06 (q, J=7.4 Hz, 4H), 1.83-1.73 (m, 3H), 1.71-1.63 (m, 1H), 1.54-1.43 (m, 3H). .sup.13C NMR (151 MHz, CD.sub.3OD) =174.6, 164.5, 146.1, 145.4, 143.6, 137.7, 137.2, 130.5, 125.9, 123.4, 121.1, 117.8, 110.6, 61.8, 60.2, 55.5, 47.7, 47.6, 39.8, 3.2, 34.3, 32.6, 30.14, 28.2, 27.9, 25.3, 25.2. HRMS (ESI) calcd. for C.sub.32H.sub.39N.sub.8O.sub.5S.sub.2[M+H] 679.2479, found 679.2456.

Example 23: N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-phenethyl)-2-methoxybenzamide

[0252] ##STR00096##

[0253] To a solution of 4-(2-aminoethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-benzenesulfonamide (Intermediate 1-23; 50 mg, 125.2 umol) in DMF (1 mL) was added 2-methoxybenzoic acid (21 mg, 137.7 umol), HATU (57 mg, 150.2 umol) and triethylamine (25 mg, 250 umol). The mixture was stirred at 25 C. for 1 hour before the reaction was quenched with water (5 mL) and extracted into dichloromethane (315 mL). The combined organic layers were washed with brine (315 mL), dried (Na.sub.2SO.sub.4), filtered and the volatiles removed under reduced pressure. The residue was purified by prep-HPLC to afford the title compound as a white (5.3 mg, 8% yield).

[0254] .sup.1H NMR (CD.sub.3OD) 7.97-7.95 (d, 2H), 7.92-7.90 (dd, 1H), 7.48-7.43 (m, 3H), 7.06-7.03 (m, 2H), 6.87 (s, 1H), 3.77 (s, 3H), 3.74-3.70 (t, 2H), 3.01 (t, 2H), 2.82-2.78 (m, 4H), 2.69-2.65 (m, 4H) and 1.00-1.92 (m, 4H).

[0255] LCMS: m/z 534.2 (M+H).sup.+

Example 24: N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)sulfamoyl)-phenethyl)-2,3-dihydrobenzofuran-7-carboxamide

[0256] ##STR00097##

[0257] Prepared as described for N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethyl)-2-methoxybenzamide (Example 23) to afford the title compound (17%) as a white solid.

[0258] .sup.1H NMR (CD.sub.3OD) 8.00-7.98 (d, 2H), 7.73-7.71 (d, 1H), 7.51-7.49 (d, 2H), 7.36-7.34 (d, 1H), 6.97-6.93 (m, 1H), 6.90 (s, 1H), 4.59-4.55 (t, 2H), 3.74-3.72 (m, 2H), 3.15-3.11 (m, 2H), 3.05-3.03 (m, 2H), 2.80-2.79 (t, 4H), 2.62-2.58 (t, 4H) and 1.98-1.92 (m, 4H).

[0259] LCMS: m/z 546.3 (M+H).sup.+

Example 25: N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyridine-2-carboxamide

[0260] ##STR00098##

[0261] Prepared as described for N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethyl)-2-methoxybenzamide (Example 23) to afford the title compound (17%) as a white solid.

[0262] .sup.1H NMR (CD.sub.3OD) 7.96-7.94 (d, 2H), 7.49-7.47 (m, 3H), 6.94 (s, 1H), 4.05-4.02 (t, 2H), 3.66-3.62 (t, 2H), 3.03-3.00 (m, 2H), 2.85-2.82 (m, 6H), 2.64-2.61 (m, 4H) and 2.03-1.97 (m, 8H).

[0263] LCMS: m/z 548.3 (M+H).sup.+

Example 26: 4-(2-(3-cyclopentylureido)ethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)benzenesulfonamide

[0264] ##STR00099##

[0265] To a solution of 4-(2-aminoethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-benzenesulfonamide (Intermediate 1-23; 60 mg, 150.2 umol) in THF (1.5 mL) was added isocyanatocyclopentane (17 mg, 150.2 umol). The mixture was stirred at 25 C. for 2 hours before the reaction mixture was concentrated in vacuo. The residue was purified by prep-HPLC to afford the title compound as a white solid (5.1 mg, 7% yield).

[0266] .sup.1H NMR (CD.sub.3OD) 7.93-7.91 (d, 2H), 7.38-7.36 (d, 2H), 6.90 (s, 1H), 3.98-3.95 (m, 1 H), 3.41-3.38 (m, 2H), 2.92-2.78 (m, 6H), 2.67 (t, 4H), 2.04-1.96 (m, 4H), 1.93-1.84 (m, 2H), 1.76-1.52 (m, 4H) and 1.42-1.29 (m, 2H).

[0267] LCMS: m/z 511.2 (M+H).sup.+

Example 27: 4-(2-(3-Benzylureido)ethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)benzenesulfonamide

[0268] ##STR00100##

[0269] Prepared as described for 4-(2-(3-cyclopentylureido)ethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)benzenesulfonamide (Example 26) to afford the title compound as a white solid (15%).

[0270] .sup.1H NMR (CD.sub.3OD) 7.93-7.91 (d, 2H), 7.38-7.23 (m, 7H), 6.90 (s, 1H), 4.30 (s, 2H), 3.44-3.42 (m, 2H), 2.89-2.80 (m, 6H), 2.68-2.66 (m, 4H) and 2.03-1.97 (m, 4H).

[0271] LCMS: m/z 533.3 (M+H).sup.+

Example 28: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-(3-phenethylureido)ethyl)benzenesulfonamide

[0272] ##STR00101##

[0273] Prepared as described for 4-(2-(3-cyclopentylureido)ethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)benzenesulfonamide (Example 26) to afford the title compound as a white solid (8%).

[0274] .sup.1H NMR (DMSO-d.sub.6) 7.82 (s, 1H), 7.78-7.76 (d, 2H), 7.31-7.27 (m, 4H), 7.20-7.17 (m, 3H), 6.84 (s, 1H), 5.92-5.88 (m, 2H), 3.25-3.20 (m, 4H), 2.76-2.72 (m, 6H), 2.68-2.66 (m, 2H), 2.60-2.56 (m, 4H), and 1.92-1.89 (m, 4H).

[0275] LCMS: m/z 547.2 (M+H).sup.+

Example 29: 2-Ethyl-N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethyl)pyrimidine-5-carboxamide

[0276] ##STR00102##

[0277] Prepared as described for N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethyl)-2-methoxybenzamide (Example 23) to afford the title compound as a white solid (8%).

[0278] .sup.1H NMR (DMSO-d.sub.6) 10.75 (s, 1H), 9.03 (s, 2H), 8.90-8.87 (br t, 1H), 8.03 (s, 1H), 7.86-7.84 (d, 2H), 7.48-7.46 (d, 2H), 6.90 (s, 1H), 3.55-3.53 (m, 2H), 3.01-2.88, (m, 4H), 2.76-2.74 (m, 4H), 2.54-2.51 (m, 4H), 1.92-1.88 (m, 4H) and 1.31-1.27 (t, 3H).

[0279] LCMS: m/z 534.1 (M+H).sup.+

Example 30: N-(4-(N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)-phenethyl)-4-methoxybenzamide

[0280] ##STR00103##

[0281] Prepared as described for N-(4-(N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-sulfamoyl)phenethyl)-2-methoxybenzamide (Example 23) to afford the title compound as a white solid (15%).

[0282] .sup.1H NMR (DMSO-d.sub.6) 10.80 (br, s, 1H), 8.46-7.43 (m, 1H), 8.01-8.00 (m, 1H), 7.85-7.78 (m, 4H), 7.46-7.44 (m, 2H), 7.01-6.99 (d, 2H), 6.90 (s, 1H), 3.79 (s, 3H), 3.51-3.47, (m, 2H), 2.92 (br t, 2H), 2.77-2.73 (m, 4H), 2.53-2.51 (m, 4H) and 1.94-1.89 (m, 4H).

[0283] LCMS: m/z 534.2 (M+H).sup.+

Example 31: N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-4-(2-(3-(4-methoxyphenyl)ureido)ethyl)benzenesulfonamide

[0284] ##STR00104##

[0285] Prepared as described for 4-(2-(3-cyclopentylureido)ethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)benzenesulfonamide (Example 26) to afford the title compound as a white solid (8%).

[0286] .sup.1H NMR (DMSO-d.sub.6) 8.27 (s, 1H), 7.94 (s, 1H), 7.84-7.82 (d, 2H), 7.42-7.40 (d, 2H), 7.28-7.26 (d, 2H), 6.88 (s, 1H), 6.81-6.79 (d, 2H), 6.05-6.02 (t, 1H), 3.69 (s, 3H), 3.30-3.32 (m, 2H), 2.83-2.80 (t, 2H), 2.78-2.74 (t, 4H), 2.58-2.55 (m, 4H) and 1.98-1.87 (m, 4H).

[0287] LCMS: m/z 549.2 (M+H).sup.+

ExamplesBiological Studies

NLRP3 Inhibition

[0288] The NLRP3-inhibitory effect of a number of the above compounds was tested by measuring the IL-1 secretion from murine bone marrow derived macrophages (BMDM) primed with lipopolysaccharide (LPS), treated with test compound (to a maximum concentration of 200 M) and finally stimulated with the NLRP3 stimulus ATP. The half-maximal inhibitory concentration (IC.sub.50) was determined for each compound and compared to a number of comparative examples (CE 1-10) including commercially available oral sulphonylureas (CE1-6) and the known NLRP3 inhibitor MCC 950 (CE7). The experimental protocol was as follows. The results are shown in Table 1 below.

[0289] MOUSE PRIMARY MACROPHAGE CELL CULTURE: Macrophages from C57BL/6 mice were differentiated from bone marrow as previously described (Schroder et al. Proceedings of the National Academy of Sciences, 2012, 109, E944-E953). The University of Queensland Animal Ethics Committee approved all experimental protocols involving mice. Murine bone marrow-derived macrophages (BMDM) were cultured in RPMI 1640 medium (Life Technologies) supplemented with 10% heat inactivated fetal calf serum, 2 mM GlutaMAX (Life Technologies) and 50 U/mL penicillin-streptomycin (Life Technologies) and 100 ng/mL recombinant human M-CSF (endotoxin free, expressed and purified by the University of Queensland Protein Expression Facility).

[0290] NLRP3 INFLAMMASOME ASSAY: BMDM were seeded at 110.sup.6 cells/mL in 96 well plates. The following day the overnight medium was replaced with Opti-MEM reduced serum medium (Life Technologies) and cells were stimulated with 100 ng/mL ultrapure E. coli K12 LPS (Invivogen) for 3 h. Compounds were prepared from a 10 mM stock by serial dilution in 80% DMSO/20% 10 mM ammonium bicarbonate. Compounds (0.001-200 M) or vehicle control were added to the LPS primed cells for 30 mins before stimulation with 2.5 mM adenosine 5-triphosphate disodium salt hydrate (ATP) (Sigma Aldrich) for 1 h. IL-1 levels in cell-free supernatants were analysed by ELISA (ReadySetGo! eBioscience).

[0291] NLRP3 DATA ANALYSIS: IC.sub.50 values were determined from ELISA data by performing non-linear regression analysis of inhibitor vs. normalized response (variable slope) using Prism Software (GraphPad).

TABLE-US-00002 TABLE 1 Example Structure IC.sub.50 1 [00105] +++ 2 [00106] +++ 3 [00107] +++ 4 [00108] +++ 5 [00109] +++ 6 [00110] +++ CE.sub.1 (Acetohexamide) [00111] xx CE.sub.2 (Glyburide) [00112] + CE.sub.3 (Glipizide) [00113] xx CE.sub.4 (Glisoxepide) [00114] x CE.sub.5 (Gliquidone) [00115] x CE6 (Glimepiride) [00116] x CE.sub.7 (MCC .sub.950) [00117] +++ CE8 [00118] +++ CE.sub.9 [00119] +++ CE.sub.10 [00120] +++ NLRP.sub.3 inhibitory activity (<1 M = +++, <10 M = ++, <50 M = +, >50 M = x, >200 M = xx).

[0292] As is evident from the above, surprisingly in spite of the structural differences the compounds of the invention show comparable NLRP3 inhibitory activity to the known NLRP3 inhibitor MCC 950, and show vastly superior inhibitory activity to the commercially available sulphonyl ureas of comparative examples CE 1-6. The compounds of the invention will therefore find application in the treatment of disorders associated with the over-activity of NLRP3.

[0293] To further confirm the NLRP3 inhibitory activity of compounds of the invention, additional assays were performed by measuring the IL-1 secretion, using either murine bone marrow derived macrophages (BMDM) or human monocyte derived macrophages (HMDM), in accordance with the following protocols. The results are shown in Table 2 below. As can be seen, the compounds of the invention show a high level of efficacy.

[0294] HMDM CELL CULTURE: To generate HMDM, human monocytes are isolated from buffy coat blood using Ficoll-Plaque Plus (GE Healthcare) and density centrifugation. CD14.sup.+ cell selection is performed using MACS magnetic beads (Miltenyl Biotec). Isolated CD14.sup.+ monocytes are differentiated in culture for 7 days with 10 ng/ml human CSF-1 (Miltenyl Biotec) in Iscove's modified Dulbecco's medium (IMDM) containing L-glutamine supplemented with 10% FBS and 1% penicillin/streptomycin (Life Technologies) as described by Croker et al., Immunol Cell Biol 2013, vol. 91, p. 625.

[0295] BMDM CELL CULTURE: Mouse bone marrow-derived macrophages (BMDM) were derived from bone marrow progenitors isolated from the femurs and tibias of C57BL/6 mice. Bones were flushed with medium, and bone marrow cells were cultured for 7 days in RPMI 1640 medium supplemented with 10% heat inactivated FCS, 2 mM GlutaMAX (Life Technologies), 50 U/ml penicillin-streptomycin (Life Technologies) and 150 ng/ml recombinant human M-CSF (endotoxin-free, expressed and purified by The University of Queensland Protein Expression Facility).

[0296] HMDM NLRP3 INFLAMMASOME ASSAY: HMDM are seeded at 110.sup.5/ml. The following day the overnight medium is replaced and cells are stimulated with Escherichia coli serotype 0111:B4 (Sigma Aldrich) for 3 h. Medium is removed and replaced with serum free medium (SFM) containing test compound 30 min prior to NLRP3 stimulation. Cells are then stimulated with: adenosine 5-triphosphate disodium salt hydrate (5 mM 1 h), nigericin (10 M 1 h), LeuLeu-OMe (1 mM 2 h) or MSU (200 g/ml 15 h). ATP can be sourced from Sigma Aldrich, nigericin and MSU from Invivogen and LeuLeu-Ome from Chem-Impex International.

[0297] BMDM NLRP3 INFLAMMASOME ASSAY: BMDM are seeded at 110.sup.5/ml. The following day the overnight medium is replaced and cells are stimulated with Ultrapure lipopolysaccharide from Escherichia coli K12 strain (InvivoGen) for 3 h. Medium is removed and replaced with serum free medium (SFM) containing test compound 30 min prior to NLRP3 stimulation. Cells are then stimulated with: adenosine 5-triphosphate disodium salt hydrate (1.25-5 mM 1 h), nigericin (5 M 1 h), LeuLeu-OMe (1 mM 2 h) or MSU (200 g/ml 15 h). ATP can be sourced from Sigma Aldrich, nigericin and MSU from Invivogen and LeuLeu-Ome from Chem-Impex International.

[0298] MEASUREMENT OF IL-1 SECRETION: For ELISA assays cells are seeded in 96 well plates. Supernatants are removed and analysed using ELISA kits according to the manufacturer's instructions (DuoSet R&D Systems, ReadySetGo! eBioscience, BD OptEIA, or Perkin Elmer AlphaLISA).

TABLE-US-00003 TABLE 2 Avg. Avg. IL-1 IL-1 IC.sub.50 IC.sub.50 Example Structure BMDM HMDM 7 [00121] +++ ND 8 [00122] ++ ND 10 [00123] ND +++ 11 [00124] ND +++ 12 [00125] ND +++ 13 [00126] ND +++ 14 [00127] ND ++ 15 [00128] ND +++ 19 [00129] ND +++ 20 [00130] ND ++ 21 [00131] ++ ++ 22 [00132] + ++ NLRP.sub.3 inhibitory activity (<1 M = +++, <10 M = ++, <50 M = +, >50 M = x, >200 M = xx, not determined = ND).

Insulin Secretion

[0299] To confirm the insulinotropic properties of the compounds of the invention, their ability to stimulate insulin secretion from MIN6 cells at a single concentration (10 M) was tested using the following protocol. The tests were conducted by the Garvan Institute of Medical Research, Sydney, Australia.

[0300] MIN6 INSULIN ASSAY PROTOCOL: MIN6 cells (passage 29-37) were incubated for 1 h in Krebs-Ringer buffer (KRB) containing 2.8 mM glucose. The cells were then stimulated for 1 h in either 2.8 mM glucose KRB, 20 mM glucose KRB, or 2.8 mM glucose KRB plus test compound. Supernatants were collected and then assayed using rat insulin radioimmunoassay kit (RI-13K, Merck Millipore).

[0301] The ability of the compounds of the invention to stimulate insulin secretion was compared to the commercially available insulin secretion stimulator glyburide (CE2) and to a number of NLRP3 inhibitors (CE7-10), including the inhibitor MCC-950 (CE7). The results are shown in FIG. 1.

[0302] As is evident, the compounds of the invention remarkably show comparable or even superior efficacy to glyburide (CE2), in spite of the significant structural modification to the urea substituents. Glyburide is only weakly active as an NLRP3 inhibitor however. In contrast, as discussed above, the compounds of the invention are potent NLRP3 inhibitors.

[0303] FIG. 1 further demonstrates that not all NLRP3 inhibitors possess the ability to induce insulin secretion. For example, the known NLRP3 inhibitor MCC-950 (CE7) merely results in basal levels of insulin secretion, while the same can be said for the comparative NLRP3 inhibitors CE8-10. Without wishing to be bound by theory, it is believed that the presence of a carbonyl-containing group attached to the cyclic sulphonyl substituent imparts on the compounds of the invention their ability to induce insulin secretion.

[0304] It is apparent therefore that a new class of compounds has been discovered, said compounds uniquely possessing dual activity, having the ability to both inhibit NLRP3 and stimulate insulin secretion. Such compounds offer distinct pharmacokinetic, regulatory and clinical advantages in the treatment of disorders responsive to both the inhibition of NLRP3 and the stimulation of insulin secretion, as well as in the treatment of patients with appropriate comorbid conditions.

[0305] 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.