SULFONYLUREAS AND SULFONYLTHIOUREAS AS NLRP3 INHIBITORS
20210122716 · 2021-04-29
Assignee
Inventors
- Matthew Cooper (Cambridge, GB)
- David Miller (Cambridge, GB)
- Angus MacLeod (Cambridge, GB)
- Stephen Thom (Nottingham, GB)
- Stephen St-Gallay (Nottingham, GB)
- Jonathan Shannon (Nottingham, GB)
Cpc classification
A61P29/00
HUMAN NECESSITIES
A61P31/00
HUMAN NECESSITIES
A61P25/28
HUMAN NECESSITIES
C07D405/06
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
A61P35/00
HUMAN NECESSITIES
International classification
C07D401/12
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
Abstract
The present invention relates to sulfonylureas and sulfonylthioureas comprising a monocyclic imidazolyl group. The present invention further relates to salts, solvates and prodrugs of such compounds, to pharmaceutical compositions comprising such compounds, and to the use of such compounds in the treatment and prevention of medical disorders and diseases, most especially by the inhibition of NLRP.sub.3.
Claims
1. A compound of formula (I): ##STR00085## or a pharmaceutically acceptable salt solvate or prodrug thereof, wherein: Q is selected from O or S; R.sup.1 is an imidazolyl group, wherein the imidazolyl group is unsubstituted or substituted with one or more monovalent substituents; and R.sup.2 is a cyclic group substituted at the α-position, wherein R.sup.2 may optionally be further substituted.
2. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein R.sup.1 is an imidazol-4-yl group or an imidazol-5-yl group.
3. (canceled)
4. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein R.sup.1 is an imidazol-2-yl group.
5. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein; (i) Q is O; and/or (ii) the imidazolyl group of R.sup.1 is substituted with one, two or three substituents independently selected from halo; —CN; —NO.sub.2; —N.sub.3; —R.sup.β; —OH; —OR.sup.β; —R.sup.α-halo; —R.sup.α—CN; —R.sup.α—NO.sub.2; —R.sup.α—N.sub.3; —R.sup.α—R.sup.β; —R.sup.α—OH; —R.sup.α—OR.sup.β; —SH; —SR.sup.P; —SOR.sup.β; —SO.sub.2H; —SO.sub.2R.sup.β; —SO.sub.2NH.sub.2; —SO.sub.2NHR.sup.β; —SO.sub.2N(R.sup.β).sub.2; —R.sup.α—SH; —R.sup.α—SR.sup.β; —R.sup.α—SOR.sup.β; —R.sup.α—SO.sub.2H; —R.sup.α—SO.sub.2R.sup.β; —R.sup.α—SO.sub.2NH.sub.2; —R.sup.α—SO.sub.2NHR.sup.β; —R.sup.α—SO.sub.2 N(R.sup.β).sub.2; —NH.sub.2; —NHR.sup.β; —N(R.sup.β).sub.2; —R.sup.α—NH.sub.2; —R.sup.α—NHR.sup.β; —R.sup.α—N(R.sup.β).sub.2; —CHO; —COR.sup.β; —COOH; —COOR.sup.β; —OCOR.sup.β; —R.sup.α—CHO; —R.sup.α—COR.sup.β; —R.sup.α—COOH; —R.sup.α—COOR.sup.β; or —R.sup.α—OCOR.sup.β; wherein each —R.sup.α— is independently selected from an alkylene, alkenylene or alkynylene group, wherein the alkylene, alkenylene or alkynylene group contains from 1 to 6 atoms in its backbone, wherein one or more carbon atoms in the backbone of the alkylene, alkenylene or alkynylene group may optionally be replaced by one or more heteroatoms N, O or S, and wherein the alkylene, alkenylene or alkynylene group may optionally be substituted with one or more halo and/or —R.sup.β groups; and wherein each —R.sup.β is independently selected from a C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6 alkenyl, C.sub.2-C.sub.6 alkynyl or C.sub.2-C.sub.6 cyclic group, and wherein any —R.sup.β may optionally be substituted with one or more C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 haloalkyl, C.sub.3-C.sub.7 cycloalkyl, —O(C.sub.1-C.sub.4 alkyl), —O(C.sub.1-C.sub.4 haloalkyl), —O(C.sub.3-C.sub.7 cycloalkyl), halo, —OH, —NH.sub.2, —CN, —C≡CH, oxo (═O), or 4- to 6-membered heterocyclic group.
6. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein the imidazolyl group of R.sup.1 is substituted with one monovalent substituent.
7. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein the imidazolyl group of R.sup.1 is substituted with two or three monovalent substituents.
8. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein each monovalent substituent of the imidazolyl group of R.sup.1 is independently selected from 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 selected from halo, —CN, —OH, —NH.sub.2 and oxo (═O), and wherein the saturated hydrocarbyl group may optionally include one or two heteroatoms N or O in its carbon skeleton.
9. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein each monovalent substituent of the imidazolyl group of R.sup.1 is acyclic.
10. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein R.sup.2 is an aryl or a heteroaryl group, wherein the aryl or the heteroaryl group is substituted at the α-position, and wherein R.sup.2 may optionally be further substituted, and optionally wherein: (i) R.sup.2 is an aryl or a heteroaryl group, wherein the aryl or the heteroaryl group is substituted at the α and α′ positions, and wherein R.sup.2 may optionally be further substituted; or (ii) R.sup.2 is a fused aryl or a fused heteroaryl group, wherein a first cycloalkyl, cycloalkenyl, non-aromatic heterocyclic, aryl or heteroaryl ring is fused to the aryl or heteroaryl group across the α,β positions and a second cycloalkyl, cycloalkenyl, non-aromatic heterocyclic, aryl or heteroaryl ring is fused to the aryl or heteroaryl group across the α′,β′ positions, and wherein R.sup.2 may optionally be further substituted.
11-12. (canceled)
13. A compound or a pharmaceutically acceptable salt, solvate or prodrug thereof, as claimed in claim 1, wherein R.sup.2 is a cyclic group substituted at the α-position with a monovalent heterocyclic group or a monovalent aromatic group, wherein a ring atom of the heterocyclic or aromatic group is directly attached to the α-ring atom of the cyclic group, wherein the heterocyclic or aromatic group may optionally be substituted, and wherein the cyclic group may optionally be further substituted.
14. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein R.sup.2 is a cyclic group substituted at the α and α′ positions, wherein R.sup.2 may optionally be further substituted.
15. (canceled)
16. A compound or a pharmaceutically acceptable salt solvate or prodrug thereof, as claimed in claim 1, wherein the compound is selected from the group consisting of: ##STR00086## ##STR00087## ##STR00088## ##STR00089##
17. (canceled)
18. A pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt, solvate or prodrug thereof, as claimed in claim 1, and a pharmaceutically acceptable excipient.
19. A method of treating or preventing a disease, disorder or condition in a subject, the method comprising the step of administering an effective amount of a compound or a pharmaceutically acceptable salt, solvate or prodrug thereof, as claimed in claim 1, to the subject, thereby treating or preventing the disease, disorder or condition, optionally wherein the disease, disorder or condition is responsive to NLRP3 inhibition.
20. (canceled)
21. The method as claimed in claim 19, 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; and (xvi) any disease where an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.
22. The method as claimed in claim 21, wherein the disease, disorder or condition is selected from: (i) inflammation; (ii) an auto-immune disease; (iii) cancer; (iv) a metabolic disease; (v) a cardiovascular disease; (vi) a respiratory disease; (vii) a non-infectious liver disease; (viii) a renal disease; (ix) an ocular disease; (x) a skin disease; (xi) a psychological disorder; (xii) a lymphatic condition; and/or (xiii) any disease, disorder or condition in which an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.
23. The method as claimed in claim 19, 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 (HFDS); (viii) Tumour Necrosis Factor (TNF) Receptor-Associated Periodic Syndrome (TRAPS); (ix) systemic juvenile idiopathic arthritis; (x) adult-onset Still's disease (AOSD); (xi) relapsing polychondritis; (xii) Schnitzler's syndrome; (xiii) Sweet's syndrome; (xiv) Behcet's disease; (xv) anti-synthetase syndrome; (xvi) deficiency of interleukin 1 receptor antagonist (DIRA); and (xvii) haploinsufficiency of A20 (HA20).
24. A method of inhibiting NLRP3 in a subject, the method comprising administering a compound or a pharmaceutically acceptable salt, solvate or prodrug thereof, as claimed in claim 1, to the subject thereby inhibiting NLRP3.
25. 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 a compound or a pharmaceutically acceptable salt, solvate or prodrug thereof, as claimed in claim 1, and analysing inhibition of NLRP3 or an effect of inhibition of NLRP3 in the cell or non-human animal by the compound.
26. The method as claimed in claim 19, wherein the compound is administered as a pharmaceutical composition further comprising a pharmaceutically acceptable excipient.
Description
EXAMPLES—COMPOUND SYNTHESIS
[0481] All solvents, reagents and compounds were purchased and used without further purification unless stated otherwise.
Abbreviations
[0482] 2-MeTHF 2-methyltetrahydrofuran
[0483] Ac.sub.2O acetic anhydride
[0484] AcOH acetic acid
[0485] aq aqueous
[0486] Boc tert-butyloxycarbonyl
[0487] br broad
[0488] Cbz carboxybenzyl
[0489] CDI 1,1-carbonyl-diimidazole
[0490] cone concentrated
[0491] d doublet
[0492] DABCO 1,4-diazabicyclo[2.2.2]octane
[0493] DCE 1,2-dichloroethane, also called ethylene dichloride
[0494] DCM dichloromethane
[0495] DIPEA N,N-diisopropylethylamine, also called Hünig's base
[0496] DMA dimethylacetamide
[0497] DMAP 4-dimethylaminopyridine, also called N,N-dimethylpyridin-4-amine
[0498] DME dimethoxyethane
[0499] DMF N,N-dimethylformamide
[0500] DMSO dimethyl sulfoxide
[0501] eq or equiv equivalent
[0502] (ES+) electrospray ionization, positive mode
[0503] Et ethyl
[0504] EtOAc ethyl acetate
[0505] EtOH ethanol
[0506] h hour(s)
[0507] HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate
[0508] HPLC high performance liquid chromatography
[0509] LC liquid chromatography
[0510] m multiplet
[0511] m-CPBA 3-chloroperoxybenzoic acid
[0512] Me methyl
[0513] MeCN acetonitrile
[0514] MeOH methanol
[0515] (M+H)+ protonated molecular ion
[0516] MHz megahertz
[0517] min minute(s)
[0518] MS mass spectrometry
[0519] Ms mesyl, also called methanesulfonyl
[0520] MsCl mesyl chloride, also called methanesulfonyl chloride
[0521] MTBE methyl tert-butyl ether, also called tert-butyl methyl ether
[0522] m/z mass-to-charge ratio
[0523] NaO.sup.tBu sodium tert-butoxide
[0524] NBS 1-bromopyrrolidine-2,5-dione, also called N-bromosuccinimide
[0525] NCS 1-chloropyrrolidine-2,5-dione, also called N-chlorosuccinimide
[0526] NMP N-methylpyrrolidine
[0527] .sup.1H NMR nuclear magnetic resonance (spectroscopy)
[0528] Pd(dba).sub.3 tris(dibenzylideneacetone) dipalladium(0)
[0529] Pd(dppf)Cl.sub.2 [1,1′-bis(diphenylphosphino)ferrocene] dichloropalladium(II)
[0530] PE petroleum ether
[0531] Ph phenyl
[0532] PMB p-methoxybenzyl, also called 4-methoxybenzyl
[0533] prep-HPLC preparative high performance liquid chromatography
[0534] prep-TLC preparative thin layer chromatography
[0535] PTSA p-toluenesulfonic acid
[0536] q quartet
[0537] RP reversed phase
[0538] RT room temperature
[0539] s singlet
[0540] Sept septuplet
[0541] sat saturated
[0542] SCX solid supported cation exchange (resin)
[0543] t triplet
[0544] T3P propylphosphonic anhydride
[0545] TBME tert-butyl methyl ether, also called methyl tert-butyl ether
[0546] TEA triethylamine
[0547] TFA 2,2,2-trifluoroacetic acid
[0548] THF tetrahydrofuran
[0549] TLC thin layer chromatography
[0550] wt % weight percent or percent by weight
[0551] Experimental Methods
[0552] Nuclear Magnetic Resonance
[0553] .sup.1H 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: [0554] a Bruker Avance III spectrometer at 400 MHz fitted with a BBO 5 mm liquid probe, [0555] a Bruker 400 MHz spectrometers using ICON-NMR, under TopSpin program control, [0556] a Bruker Avance III HD spectrometer at 500 MHz, equipped with a Bruker 5 mm SmartProbe™, [0557] 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 [0558] an Agilent MercuryPlus 300 instrument fitted with a 7.05 Tesla magnet from Oxford instruments, 4 nuclei auto-switchable probe and Mercury plus console.
[0559] LC-MS
[0560] 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 2.1×30 mm, 5 μm.
[0561] Reversed Phase HPLC Conditions for the LCMS Analytical Methods
[0562] Methods 1a and 1b: Waters Xselect CSH C18 XP column (4.6×30 mm, 2.5 μm) at 40° C.; flow rate 2.5-4.5 mL min.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.
[0563] Method 1c: Agilent 1290 series with UV detector and HP 6130 MSD mass detector using Waters XBridge BEH C18 XP column (2.1×50 mm, 2.5 μm) at 35° C.; flow rate 0.6 mL/min; mobile phase A: ammonium acetate (10 mM); water/MeOH/acetonitrile (900:60:40); mobile phase B: ammonium acetate (10 mM); water/MeOH/acetonitrile (100:540:360); over 4 min employing UV detection at 215 and 238 nm. 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.
[0564] Reversed Phase HPLC Conditions for the UPLC Analytical Methods
[0565] Methods 2a and 2b: Waters BEH C18 (2.1×30 mm, 1.7 μm) at 40° C.; flow rate 0.77 mL min.sup.−1 eluted with a H.sub.2O-MeCN gradient containing either 0.1% v/v formic acid (Method 2a) or 10 mM NH.sub.4HCO.sub.3 in water (Method 2b) over 3 min employing UV detection at 254 nm. 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.
[0566] Preparative Reversed Phase HPLC General Methods
[0567] Method 1 (acidic preparation): Waters X-Select CSH column C18, 5 μm (19×50 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.
[0568] Method 2 (basic preparation): Waters X-Bridge Prep column C18, 5 μm (19×50 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.
[0569] Method 3: Phenomenex Gemini column, 10 μm (150×25 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.
[0570] Method 4: Revelis C18 reversed-phase 12 g cartridge [carbon loading 18%; surface area 568 m.sup.2/g; pore diameter 65 Angstrom; pH (5% slurry) 5.1; average particle size 40 μm], 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 P1: (R)-1-(2-Hydroxypropyl)-1H-imidazole-4-sulfonamide
Step A: N,N-Bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide
[0571] ##STR00033##
[0572] A solution of 1H-imidazole-4-sulfonyl chloride (2.5 g, 15.01 mmol) in DCM (10 mL) was added slowly to a solution of bis(4-methoxybenzyl)amine (4 g, 15.54 mmol) and Et.sub.3N (4.5 mL, 32.3 mmol) in DCM (50 mL) cooled in an ice bath. The mixture was stirred for 30 minutes, warmed to room temperature and stirred for 2 hours. The DCM was removed under pressure and replaced with dioxane (50 mL) and the mixture heated under reflux for 48 hours, cooled, and then partitioned between EtOAc (200 mL) and water (200 mL). The organic layer was dried (MgSO.sub.4), filtered and evaporated to give an oil that was purified by chromatography on silica gel (120 g column, 0-100% EtOAc/isohexane). The product was triturated in TBME/EtOAc, filtered and dried to afford the title compound (2.864 g, 48%) as a solid.
[0573] .sup.1H NMR (CDCl.sub.3) δ 7.92 (d, J=1.3 HZ, 1H), 7.52 (d, J=1.3 HZ, 1H), 7.06-7.02 (m, 4H), 6.79-6.75 (m, 4H), 4.30 (s, 4H), 3.77 (s, 6H). Exchangeable proton not visible.
[0574] LCMS; m/z 388 (M+H).sup.+ (ES.sup.+); 386 (M−H).sup.− (ES.sup.−).
Step B: (R)-1-(2-Hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide
[0575] ##STR00034##
[0576] A mixture of N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (500 mg, 1.29 mmol), (R)-2-methyloxirane (0.18 ml, 2.57 mmol) and K.sub.2CO.sub.3 (535 mg, 3.87 mmol) in acetonitrile (10 mL) was stirred at 60° C. overnight. Upon cooling to room temperature, the reaction mixture was diluted with H.sub.2O (20 mL) and extracted with EtOAc (3×50 mL). The combined organic extracts were washed with brine (30 mL), passed through a phase separator, and concentrated in vacuo. The residue was loaded onto silica and purified by chromatography on silica gel (40 g column, 0-100% EtOAc/isohexane) to afford the title compound (468 mg, 68%) as a clear colourless oil.
[0577] .sup.1H NMR (DMSO-d.sub.6) δ 7.82 (d, J=1.3 Hz, 1H), 7.81 (d, J=1.3 Hz, 1H), 7.03 (d, J=8.6 Hz, 4H), 6.80 (d, J=8.6 Hz, 4H), 4.18 (s, 4H), 4.06-4.00 (m, 2H), 3.95-3.86 (m, 2H), 3.71 (s, 6H), 1.04 (d, J=5.9 Hz, 3H).
[0578] LCMS; m/z 446.4 (M+H).sup.+ (ES.sup.+).
Step C: (R)-1-(2-Hydroxypropyl)-1H-imidazole-4-sulfonamide
[0579] ##STR00035##
[0580] A mixture of (R)-1-(2-hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (460 mg, 0.857 mmol) and TFA (5.5 mL) was stirred at room temperature for 2 days. The reaction mixture was concentrated in vacuo and the residue loaded onto silica and purified by chromatography on silica gel (12 g column, 0-10% MeOH/DCM) to afford a clear colourless oil, which was further purified by prep-HPLC to afford the title compound (123 mg, 68%) as a clear colourless oil.
[0581] .sup.1H NMR (DMSO-d.sub.6) δ 7.71 (d, J=1.3 Hz, 1H), 7.59 (d, J=1.3 Hz, 1H), 7.12 (s, 2H), 5.00 (d, J=4.1 Hz, 1H), 4.07-3.96 (m, 1H), 3.93-3.79 (m, 2H), 1.03 (d, J=6.0 Hz, 3H).
[0582] LCMS; m/z 206.2 (M+H).sup.+ (ES.sup.+).
Intermediate P2: (S)-1-(2-Hydroxypropyl)-1H-imidazole-4-sulfonamide
Step A: (S)-1-(2-Hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide
[0583] ##STR00036##
[0584] Prepared according to the general procedure of (R)-1-(2-hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (Intermediate P1, Step B) from N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (Intermediate P1, Step A) and (S)-2-methyloxirane to afford the title compound (160 mg, 37%) as a clear colourless oil.
[0585] .sup.1H NMR (DMSO-d.sub.6) δ 7.81 (d, J=1.2 Hz, 1H), 7.80 (d, J=1.2 Hz, 1H), 7.02 (d, J=8.7 Hz, 4H), 6.80 (d, J=8.7 Hz, 4H), 5.04 (d, J=4.6 Hz, 1H), 4.18 (s, 4H), 4.06-3.99 (m, 1H), 3.97-3.84 (m, 2H), 3.71 (s, 6H), 1.04 (d, J=5.9 Hz, 3H).
[0586] LCMS; m/z 446.2 (M+H).sup.+ (ES.sup.+).
Step B: (S)-1-(2-Hydroxypropyl)-1H-imidazole-4-sulfonamide
[0587] ##STR00037##
[0588] Prepared according to the general procedure of (R)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Intermediate P1, Step C) from (S)-1-(2-hydroxypropyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide to afford the title compound (56 mg, 70%) as a clear colourless oil.
[0589] .sup.1H NMR (DMSO-d.sub.6) δ 7.71 (d, J=1.3 Hz, 1H), 7.59 (d, J=1.3 Hz, 1H), 7.11 (s, 2H), 5.00 (d, J=4.6 Hz, 1H), 4.04-3.95 (m, 1H), 3.92-3.80 (m, 2H), 1.03 (d, J=6.0 Hz, 3H).
[0590] LCMS; m/z 206.2 (M+H).sup.+ (ES.sup.+).
Intermediate P3: 1-(2-(Dimethylamino)ethyl)-1H-imidazole-4-sulfonamide
Step A: N,N-Bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide
[0591] ##STR00038##
[0592] A solution of 1H-imidazole-4-sulfonyl chloride (2.5 g, 15.01 mmol) in DCM (10 mL) was added slowly to a solution of bis(4-methoxybenzyl)amine (4 g, 15.54 mmol) and Et.sub.3N (4.5 mL, 32.3 mmol) in DCM (50 mL) cooled in an ice bath. The mixture was stirred for 30 minutes, warmed to room temperature and stirred for 2 hours. The DCM was removed under pressure and replaced with dioxane (50 mL). Then the reaction mixture was heated under reflux for 48 hours, cooled and partitioned between EtOAc (200 mL) and water (200 mL). The organic layer was dried (MgSO.sub.4), filtered and evaporated to give an oil that was purified by chromatography on silica gel (120 g column, 0-100% EtOAc/isohexane). The product was triturated in TBME/EtOAc, filtered and dried to afford the title compound (2.864 g, 48%) as a solid.
[0593] .sup.1H NMR (CDCl.sub.3) δ 7.92 (d, J=1.3 HZ, 1H), 7.52 (d, J=1.3 HZ, 1H), 7.06-7.02 (m, 4H), 6.79-6.75 (m, 4H), 4.30 (s, 4H), 3.77 (s, 6H). Exchangeable proton not visible.
[0594] LCMS; m/z 388 (M+H).sup.+ (ES.sup.+); 386 (M−H).sup.− (ES.sup.−).
Step B: 1-(2-Hydroxyethyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide
[0595] ##STR00039##
[0596] A mixture of N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (1 g, 2.58 mmol), oxirane (2.5 M in THF) (2 mL, 5.00 mmol) and K.sub.2CO.sub.3 (1.07 g, 7.74 mmol) in acetonitrile (20 mL) was stirred at 50° C. for 3 days. Upon cooling to room temperature, the reaction mixture was diluted with H.sub.2O (40 mL) and extracted with EtOAc (3×80 mL). The combined organic extracts were washed with brine (50 mL), passed through a phase separator and the solvent was removed in vacuo. The residue was loaded onto silica and purified by chromatography on silica gel (40 g column, 0-100% EtOAc/isohexane, eluting at 100%) to afford the title compound (679 mg, 61%) as a clear colourless solid.
[0597] .sup.1H NMR (DMSO-d.sub.6) δ 7.85 (d, J=1.3 Hz, 1H), 7.84 (d, J=1.3 Hz, 1H), 7.03 (d, J=8.7 Hz, 4H), 6.80 (d, J=8.7 Hz, 4H), 5.04 (t, J=5.1 Hz, 1H), 4.18 (s, 4H), 4.08 (t, J=5.3 Hz, 2H), 3.71 (s, 6H), 3.70-3.66 (m, 2H).
[0598] LCMS; m/z 432.4 (M+H).sup.+ (ES.sup.+).
Step C: 1-(2-(Dimethylamino)ethyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide
[0599] ##STR00040##
[0600] To a solution of 1-(2-hydroxyethyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (675 mg, 1.564 mmol) in DCM (8 mL) at 0° C. was added DIPEA (0.41 mL, 2.348 mmol) and methanesulfonyl chloride (0.16 mL, 2.053 mmol). The reaction mixture was warmed to room temperature and stirred for 2 hours before being quenched by addition of aqueous NaHCO.sub.3 (10 mL). The reaction mixture was extracted twice with DCM (15 mL) and the combined organic extracts were passed through a phase separator and concentrated in vacuo. The orange residue was dissolved in THF (8 mL), dimethylamine (2M in THF) (2.4 mL, 4.80 mmol) and potassium iodide (130 mg, 0.782 mmol) were added, and the reaction mixture was heated to 60° C. and stirred overnight. Additional dimethylamine (2M in THF) (2.4 mL, 4.80 mmol) was added and stirring was continued overnight. The reaction mixture was diluted with aqueous NaHCO.sub.3 (20 mL) and extracted with EtOAc (3×40 mL). The combined organic extracts were passed through a phase separator and the solvent was removed in vacuo. The residue was dissolved in MeOH (30 mL), SCX (˜12 g) was added and the suspension was stirred at room temperature for 30 minutes. The mixture was transferred into a cartridge, sequentially washed with DCM/MeOH (9:1) and MeOH, and the product was eluted with 0.7 M NH.sub.3 in MeOH to afford the title compound (585 mg, 73%) as a yellow oil.
[0601] .sup.1H NMR (DMSO-d.sub.6) δ 7.87 (s, 2H), 7.02 (d, J=8.7 Hz, 4H), 6.79 (d, J=8.7 Hz, 4H), 4.18 (s, 4H), 4.12 (t, J=6.2 Hz, 2H), 3.71 (s, 6H), 2.58 (t, J=6.2 Hz, 2H), 2.18 (s, 6H).
[0602] LCMS; m/z 459.0 (M+H).sup.+ (ES.sup.+).
Step D: 1-(2-(Dimethylamino)ethyl)-1H-imidazole-4-sulfonamide
[0603] ##STR00041##
[0604] A mixture of 1-(2-(dimethylamino)ethyl)-N,N-bis(4-methoxybenzyl)-1H-imidazole-4-sulfonamide (585 mg, 1.135 mmol) and TFA (4 mL, 62.8 mmol) was stirred at room temperature overnight. The mixture was evaporated and the residue was dissolved in MeOH (30 mL) and DCM (10 mL). SCX (8 g) was added and the mixture was stirred for 30 minutes at room temperature, transferred to a cartridge and the solid washed sequentially with DCM:MeOH (9:1) and MeOH. The product was eluted with 0.7 M NH.sub.3 in MeOH to give crude product, which was further purified by chromatography on silica gel (24 g column, 0-10% (0.7 M ammonia/MeOH/DCM) to afford the title compound (180 mg, 72%) as a pale yellow oil.
[0605] .sup.1H NMR (DMSO-d.sub.6) δ 7.77 (d, J=1.4 Hz, 1H), 7.66 (d, J=1.3 Hz, 1H), 7.11 (s, 2H), 4.09 (t, J=6.1 Hz, 2H), 2.56 (t, J=6.1 Hz, 2H), 2.17 (s, 6H).
[0606] LCMS; m/z 219.3 (M+H).sup.+ (ES.sup.+).
Intermediate P4: (4-(Dimethylamino)pyridin-1-ium-1-carbonyl)((1-isopropyl-1H-imidazol-4-yl)sulfonyl)amide
[0607] ##STR00042##
[0608] A solution of 1-iso-propyl-1H-imidazole-4-sulfonamide (161 mg, 0.851 mmol) in acetonitrile (1 mL) was treated with N,N-dimethylpyridin-4-amine (208 mg, 1.702 mmol) and the reaction mixture was stirred at room temperature until sulfonamide had dissolved. Then diphenyl carbonate (200 mg, 0.936 mmol) was added and the reaction mixture was left for 16 hours at room temperature. The resulting precipitate was separated by filtration, washed with methyl tert-butylether and dried to afford the title compound (186 mg, 65%) as a white solid which was used without further purification.
Intermediate P5: 2-((Dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide, and
Intermediate P6: 5-((Dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide
Step A: N,N-Bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide
[0609] ##STR00043##
[0610] 1-Methyl-1H-imidazole-4-sulfonyl chloride (4 g, 22.15 mmol) was added portionwise to a solution of bis(4-methoxybenzyl)amine (5.98 g, 23.25 mmol) and triethylamine (6.17 ml, 44.3 mmol) in DCM (100 mL) cooled in an ice bath. The mixture was stirred for 30 minutes, warmed to room temperature and stirred for 16 hours. The mixture was washed with water (2×50 mL), aq 1 M HCl (50 mL), water (50 mL), and brine (50 mL), then dried over MgSO.sub.4, filtered and evaporated to dryness to give crude product as a cream coloured solid. The crude product was dissolved in a minimum amount of DCM to load, then purified by column chromatography on silica gel (220 g column, 0-80% EtOAc/isohexane) to afford the title compound (5.1 g, 55%) as a white solid.
[0611] .sup.1H NMR (DMSO-d.sub.6) δ 7.87-7.78 (m, 2H), 7.09-6.99 (m, 4H), 6.91-6.73 (m, 4H), 4.19 (s, 4H), 3.72 (s, 6H), 3.33 (s, 3H).
[0612] LCMS; m/z 402.3 (M+H).sup.+ (ES.sup.+).
Step B: 2-((Dimethylamino)methyl)-N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide and 5-((dimethylamino)methyl)-N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide
[0613] ##STR00044##
[0614] A solution of n-BuLi (2.5M in hexanes) (1.046 mL, 2.62 mmol) was added dropwise to a stirred solution of N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide (1 g, 2.491 mmol) in THF (14 mL) at −78° C. The reaction was stirred for 1 hour and then N-methyl-N-methylenemethanaminium iodide (0.922 g, 4.98 mmol) was added. The reaction mixture was left at −78° C. for 1 hour. The reaction was quenched with water (20 mL) and extracted with EtOAc (2×20 mL). The combined organic layers were dried over MgSO.sub.4, filtered and concentrated to dryness. The crude product was purified by chromatography on silica gel (24 g column, 0-5% MeOH/DCM) to afford an inseparable 85:15 mixture of 2-((dimethylamino)methyl)-N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide and 5-((dimethylamino)methyl)-N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide (374 mg, 27.8%) as a yellow oil.
[0615] .sup.1H NMR (major product) (DMSO-d.sub.6) δ 7.83 (s, 1H), 7.05-6.98 (m, 4H), 6.83-6.74 (m, 4H), 4.24 (s, 4H), 3.71 (s, 6H), 3.70 (s, 3H), 3.68 (s, 2H), 2.16 (s, 6H).
[0616] m/z 459.4 (M+H).sup.+ (ES.sup.+).
Step C: 2-((Dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide & 5-((dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide
[0617] ##STR00045##
[0618] A 85:15 mixture of 2-((dimethylamino)methyl)-N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide and 5-((dimethylamino)methyl)-N,N-bis(4-methoxybenzyl)-1-methyl-1H-imidazole-4-sulfonamide (552 mg, 1.204 mmol) was dissolved in TFA (5 ml, 1.204 mmol) and stirred overnight. Additional TFA (2 mL) was added and stirred for a further 24 hours at room temperature. The mixture was concentrated in vacuo and the residue was suspended in a mixture of MeOH (50 mL) and DCM (10 mL). SCX (3 eq.) was added and the suspension was stirred at room temperature for 1 hour. The SCX was filtered and washed with MeOH (50 mL) and the product was then eluted with 0.7% ammonia in MeOH (50 mL). After concentration in vacuo, a 85:15 mixture of 2-((dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide and 5-((dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide (222 mg, 84%) was isolated as a pale yellow oil.
[0619] .sup.1H NMR (major product) (DMSO-d.sub.6) δ 7.72 (s, 1H), 7.10 (s, 2H), 3.66 (s, 3H), 3.65 (s, 2H), 2.15 (s, 6H).
Intermediate P7: 2-Isopropyl-1-methyl-1H-imidazole-5-sulfonamide, and
Intermediate P8: 2-Isopropyl-1-methyl-1H-imidazole-4-sulfonamide
Step A: 2-Isopropyl-1-methyl-1H-imidazole-5-sulfonyl chloride and 2-isopropyl-1-methyl-1H-imidazole-4-sulfonyl chloride
[0620] ##STR00046##
[0621] To chlorosulfonic acid (1.2 mL, 24.1 mmol) stirred in a microwave vial (20 mL) at room temperature was added 1-methyl-2-(propan-2-yl)-1H-imidazole (1.0 g, 8.0 mmol). The microwave vial was sealed and heated in a sand bath at 150° C. for 3 hours. The reaction mixture was cooled to room temperature and then thionyl chloride (0.7 mL, 9.1 mmol) was added. The reaction mixture was heated at 100° C. overnight and then cooled to room temperature and poured into ice. The aqueous mixture was neutralised with sodium bicarbonate to pH 5, and then extracted with DCM. The organic layer was washed brine, dried over Na.sub.2SO.sub.4, filtered and then concentrated in vacuo. The crude product was dissolved in DCM and then submitted for normal phase flash chromatography on silica using heptane and EtOAc as eluent to afford 2-isopropyl-1-methyl-1H-imidazole-5-sulfonyl (157 mg, 9%) and 2-isopropyl-1-methyl-1H-imidazole-4-sulfonyl chloride (130 mg, 7%).
[0622] .sup.1H NMR for 2-isopropyl-1-methyl-1H-imidazole-5-sulfonyl chloride:
[0623] .sup.1H NMR (CDCl.sub.3) δ 7.55 (s, 1H), 3.70 (s, 3H), 3.12-2.95 (m, 1H), 1.37 (d, 6H).
[0624] .sup.1H NMR for 2-isopropyl-1-methyl-1H-imidazole-4-sulfonyl chloride:
[0625] .sup.1H NMR (CDCl.sub.3) δ 7.78 (s, 1H), 3.88 (s, 3H), 3.18-2.96 (m, 1H), 1.38 (d, 6H).
Step B1: 2-Isopropyl-1-methyl-1H-imidazole-5-sulfonamide
[0626] ##STR00047##
[0627] To a solution of 2-isopropyl-1-methyl-1H-imidazole-5-sulfonyl chloride (Intermediates P7 & P8, Step A) (130 mg, 0.58 mmol) in DCM (5 mL) was added 7M NH.sub.3 in methanol (0.32 mL, 2.32 mmol). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The crude product was dissolved in methanol, coated on hydromatrix and then submitted for normal phase flash chromatography on silica using DCM and a mixture of 3.5 M NH.sub.3 in methanol as eluent to afford the title compound (41 mg, 34%).
[0628] .sup.1H NMR (DMSO-d.sub.6) δ 7.62 (s, 2H), 7.22 (s, 1H), 3.71 (s, 3H), 3.19-3.05 (m, 1H), 1.21 (d, 6H).
Step B2: 2-Isopropyl-1-methyl-1H-imidazole-4-sulfonamide
[0629] ##STR00048##
[0630] To a solution of 2-isopropyl-1-methyl-1H-imidazole-4-sulfonyl chloride (Intermediates P7 & P8, Step A) (157 mg, 0.70 mmol) in DCM (7 mL) was added 7M NH.sub.3 in methanol (0.4 mL, 2.8 mmol). The reaction mixture was stirred at room temperature for 3 hours and then extra 7M NH.sub.3 in methanol (0.4 mL, 2.8 mmol) was added. The mixture was stirred for an additional 4 hours and then concentrated in vacuo. The crude product was dissolved in methanol, coated on hydromatrix and then submitted for normal phase flash chromatography on silica using DCM and a mixture of 3.5 M NH.sub.3 in methanol as eluent to afford the title compound (85 mg, 59%).
[0631] .sup.1H NMR (DMSO-d.sub.6) δ 7.50 (s, 1H), 7.05 (s, 2H), 3.62 (s, 3H), 3.09 (p, 1H), 1.21 (d, 6H).
Intermediate P9: 1-Isopropyl-2-methyl-1H-imidazole-4-sulfonamide
Step A: N,N-Bis(4-Methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide
[0632] ##STR00049##
[0633] 2-Methyl-1H-imidazole-4-sulfonylchloride (1 g, 5.5 mmol) was suspended in DCM (30 mL) at room temperature. To this suspension was added bis(4-methoxybenzyl)amine (1.5 g, 6 mmol) and potassium tert-butoxide (0.25 g, 2 mmol). The reaction mixture was stirred for 18 hours at room temperature, washed with water and brine, dried (Na.sub.2SO.sub.4), filtered and evaporated. The residue was further purified over silica (using a gradient of methanol in DCM 0-5% as eluent) to afford the title compound as a brown oil (2.4 g, 100%).
[0634] .sup.1H NMR (CDCl.sub.3) δ 7.37 (s, 1H), 7.07 (d, 4H), 6.76 (d, 4H), 4.30 (s, 4H), 3.80 (s, 6H), 2.43 (s, 3H).
Step B: 1-Isopropyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide
[0635] ##STR00050##
[0636] N, N-Bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (250 mg, 0.63 mmol) was dissolved in DMF (10 mL). Potassium carbonate (500 mg, 3.6 mmol), followed by 2-iodopropane (0.42 g, 2.5 mmol) were added. The reaction mixture was stirred for 48 hours at room temperature and then diluted with DCM and washed with water and brine. The organic layer was dried (Na.sub.2SO.sub.4), filtered and evaporated. The residue was purified over silica (using a gradient of methanol in DCM 0-5% as eluent) to afford the title compound (283 mg, 100%).
[0637] .sup.1H NMR (CDCl.sub.3) δ 7.37 (s, 1H), 7.10 (d, 4H), 6.76 (d, 4H), 4.32 (s, 4H), 3.78 (s, 6H), 2.43 (s, 3H) 1.42 (d, 6H).
Step C: 1-Isopropyl-2-methyl-1H-imidazole-4-sulfonamide
[0638] ##STR00051##
[0639] 1-Isopropyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (0.28 g, 0.64 mmol) was dissolved in DCM (10 mL). Trifluoroacetic acid (10 mL) was added and the mixture was stirred for 18 hours at room temperature. The solvents were evaporated and to the residue was added NH.sub.3 (7 M) in methanol. The solvents were evaporated and the residue was triturated with water, filtered and lyophilized. The residue was further purified over silica, using DCM and a mixture of 3.5 M NH.sub.3 in methanol as the eluent to afford the title compound as a white solid (60 mg, 46%).
[0640] .sup.1H NMR (CDCl.sub.3) δ 748 (s, 1H), 5.03 (s, br, 2H), 4.32 (m, 1H), 2.43 (s, 3H), 1.45 (d, 6H).
Intermediate P10: 1-Ethyl-2-methyl-1H-imidazole-4-sulfonamide
Step A: 1-Ethyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide
[0641] ##STR00052##
[0642] Prepared as described for 1-isopropyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step B), using N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step A) (250 mg, 0.63 mmol) and ethyl iodide (390 mg, 2.5 mmol) to afford the title compound (280 mg, 100%).
[0643] .sup.1H NMR (CDCl.sub.3) δ 7.32 (s, 1H), 7.10 (d, 4H), 6.76 (d, 4H), 4.32 (s, 4H), 3.89 (q, 2H), 3.78 (s, 6H), 243 (s, 3H), 142 (t, 3H).
Step B: 1-Ethyl-2-methyl-1H-imidazole-4-sulfonamide
[0644] ##STR00053##
[0645] Prepared as described for 1-isopropyl-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step C) using 1-ethyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide to afford the title compound (0.28 g, 14%) as a white solid.
[0646] .sup.1H NMR (CD3OD) δ 7.75 (s, 1H), 4.00 (q, 2H), 2.39 (s, 3H), 1.39 (t, 3H).
Intermediate P11: 2-Methyl-1-((tetrahydrofuran-2-yl)methyl)-1H-imidazole-4-sulfonamide
Step A: N,N-Bis(4-methoxybenzyl)-2-methyl-1-((tetrahydrofuran-2-yl)methyl)-1H-imidazole-4-sulfonamide
[0647] ##STR00054##
[0648] Prepared as described for 1-isopropyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step B), using N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step A) (0.28 g, 0.64 mmol) and 3-bromomethylfuran (400 mg, 2.5 mmol) to afford the title compound (242 mg, 78%) as a solid.
[0649] .sup.1H NMR (CDCl.sub.3) δ 7.45 (s, 1H), 7.10 (d, 4H), 6.76 (d, 4H), 4.32 (s, 4H), 3.97 (dd, 1H), 3.82 (m, 2H), 3.78 (s, 6H), 2.44 (s, 3H), 2.07 (m, 1H), 1.89 (m, 2H), 1.61 (m, 1H).
Step B: 2-Methyl-1-((tetrahydrofuran-2-yl)methyl)-1H-imidazole-4-sulfonamide
[0650] ##STR00055##
[0651] Prepared as described for 1-isopropyl-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step C) using N,N-bis(4-methoxybenzyl)-2-methyl-1-((tetrahydrofuran-2-yl)methyl)-1H-imidazole-4-sulfonamide (0.2 g, 042 mmol) to afford the title compound (45 mg, 45%) as a white solid.
[0652] .sup.1H NMR (CD3OD): δ 7.56 (s, 1H), 4.11 (m, 2H), 3.96 (dd, 1H), 3.80 (m, 2H), 2.40 (s, 3H), 2.07 (m, 1H), 1.89 (m, 2H), 1.61 (m, 1H).
Intermediate P12: 1-(2-Hydroxyethyl)-2-methyl-1H-imidazole-4-sulfonamide
Step A: 1-(2-((tert-Butyldimethylsilyl)oxy)ethyl)-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide
[0653] ##STR00056##
[0654] Prepared as described for 1-isopropyl-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step B) using N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step A) (0.28 g, 0.64 mmol) and (2-bromoethoxy)(tert-butyl)dimethylsilane (600 mg, 2.5 mmol) to afford the title compound (350 mg, 100%).
[0655] .sup.1H NMR (CDCl.sub.3) δ 7.40 (s, 1H), 7.10 (d, 4H), 6.76 (d, 4H), 4.29 (s, 4H), 3.97 (t, 2H), 3.86 (t, 2H), 3.78 (s, 6H), 2.45 (s, 3H), 0.84 (s, 9H), −0.03 (s, 6H).
Step B: 1-(2-Hydroxyethyl)-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide
[0656] ##STR00057##
[0657] 1-(2-((tert-Butyldimethylsilyl)oxy)ethyl)-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (350 mg, 0.63 mmol) was dissolved in DCM (10 mL). Tetrabutylammonium fluoride (0.60 g, 1.90 mmol) was added and the mixture was stirred overnight at room temperature. The DCM layer was washed with brine, dried, filtered and evaporated to afford the title compound, which was used as such for the next step.
Step C: 1-(2-Hydroxyethyl)-2-methyl-1H-imidazole-4-sulfonamide
[0658] ##STR00058##
[0659] Prepared as described for 1-isopropyl-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9, Step C) using 1-(2-hydroxyethyl)-N,N-bis(4-methoxybenzyl)-2-methyl-1H-imidazole-4-sulfonamide (0.28 g, 0.64 mmol) to afford the title compound (340 mg, 100%) as a solid, still containing inorganic salts.
[0660] .sup.1H NMR (CD3OD) δ 7.59 (s, 1H), 4.07 (t, 2H), 3.81 (t, 2H), 2.42 (s, 3H).
Intermediate A1: 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene
[0661] ##STR00059##
[0662] To a solution of phosgene (4.45 mL, 20% weight in toluene, 8.4 mmol) in EtOAc (90 mL) was added dropwise a solution of 1,2,3,5,6,7-hexahydro-s-indacen-4-amine (589 mg, 3.4 mmol) in EtOAc (45 mL) at ambient temperature. The resulting reaction mixture was then heated to reflux for 3 hours and upon cooling was filtered and concentrated in vacuo to afford the title compound as a brown oil (756 mg, 100%). The crude product was used directly in the next step without further purification.
[0663] .sup.1H NMR (CDCl.sub.3) δ 6.8 (s, 1H), 2.89 (m, 8H) and 2.09 (m, 4H).
Intermediate A2: 2-Isocyanato-1,3-diisopropylbenzene
[0664] ##STR00060##
[0665] 2,6-Diisopropylaniline (3.07 g, 17.14 mmol) was dissolved in dry THF (40 mL) and Et.sub.3N (3 mL, 21.52 mmol) was added. A solution of triphosgene (4.26 g, 14.35 mmol) in dry THF (12 mL) was added over 5 minutes, resulting in the formation of a thick colourless precipitate. The reaction mixture was stirred at room temperature overnight. The THF was removed in vacuo and toluene (50 mL) was added. The mixture was filtered through a short silica plug eluting with toluene (150 mL). The filtrate was concentrated in vacuo to afford the title compound (2.76 g, 92%) as a colourless oil.
[0666] .sup.1H NMR (CDCl.sub.3) δ 7.20-7.10 (m, 3H), 3*22 (hept, J=6.9 Hz, 2H), 1.26 (d, J=6.8 Hz, 12H).
Intermediate A2: 4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)aniline
Step A: 2-Bromo-4-fluoro-6-iso-propylaniline
[0667] ##STR00061##
[0668] N-Bromosuccinimide (5.64 g, 31.7 mmol) was added portionwise to 4-fluoro-2-isopropylaniline (4.62 g, 30.2 mmol) in DCM (72 mL) at 0° C. The resulting mixture was stirred at 0° C. for 1 hour and then left to warm to room temperature over 21 hours. The reaction mixture was washed with a solution of aqueous sodium hydroxide (2 M, 2×50 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to give a brown residue. The brown residue was put onto a plug of silica (50 g) and washed through with 50% DCM in iso-hexane (500 mL). The red filtrate was concentrated to dryness and the crude product was purified by chromatography on silica gel (120 g column, 0-10% DCM/iso-hexane) to afford the title compound (4.99 g, 70%) as a red oil.
[0669] .sup.1H NMR (CDCl.sub.3) δ 7.07 (dd, 1H), 6.86 (dd, 1H), 4.14 (s, 2H), 2.93 (sep, 1H) and 1.25 (d, 6H).
[0670] LCMS m/z 232.2/234.3 (M+H).sup.+ (ES.sup.+).
Step B: 4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)aniline
[0671] ##STR00062##
[0672] (2-Methoxypyridin-4-yl)boronic acid (144 mg, 0.938 mmol) was added to a stirred, N.sub.2-degassed mixture of 2-bromo-4-fluoro-6-isopropylaniline (200 mg, 0.853 mmol), Pd(dppf)Cl.sub.2 (31.2 mg, 0.043 mmol) and potassium carbonate (354 mg, 2.56 mmol) in 1,4-dioxane:water (10:1, 6.6 mL). The reaction mixture was then heated to 80° C. under an N.sub.2 atmosphere for 22.5 hours. The reaction mixture was left to cool to room temperature and poured onto EtOAc (10 mL) and water (5 mL). The organic layer was collected and the aqueous layer extracted with EtOAc (2×10 mL). The combined organic layers were dried (Na.sub.2SO.sub.4), filtered and evaporated to dryness. The crude product was purified by chromatography on silica gel (24 g column, 0-50% EtOAc/isohexane) to afford the title compound (174 mg, 78%) as a light brown solid.
[0673] .sup.1H NMR (CDCl.sub.3) δ 8.25 (d, J=5.3 Hz, 1H), 7.00 (dd, J=5.3, 1.4 Hz, 1H), 6.93 (dd, J=9.9, 2.9 Hz, 1H), 6.85 (s, 1H), 6.71 (dd, J=8.6, 3.0 Hz, 1H), 4.01 (s, 3H), 2.92 (hept, J=6.9 Hz, 1H), 1.28 (d, J=6.8 Hz, 6H). Exchangeable NH.sub.2 signal seen as broad hump from 4.5-0.5 ppm.
[0674] LCMS m/z 261.1 (M+H).sup.+ (ES.sup.+).
Intermediate A4: 4-Fluoro-2-isopropyl-6-(i-methyl-1H-pyrazol-4-yl)aniline
[0675] ##STR00063##
[0676] To a sealed vial was added 2-bromo-4-fluoro-6-isopropylaniline (Intermediate A3, Step A) (350 mg, 1.508 mmol) in DMF (15 mL), followed by the addition of (1-methyl-1H-pyrazol-4-yl)boronic acid (190 mg, 1.508 mmol), Pd(PPh.sub.3).sub.4 (174 mg, 0.151 mmol) and aqueous 2.0M Na.sub.2CO.sub.3 (3 mL). The reaction mixture is heated under argon at 100° C. overnight. The residue was diluted with EtOAc (20 mL), washed with brine (20 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The crude product was purified by chromatography on silica gel (40 g column, 0-60% EtOAc/isohexane) to afford the title compound (85 mg, 23%) as a brown oil.
[0677] .sup.1H NMR (CDCl.sub.3) δ 7.68 (d, 1H), 7.58 (d, 1H), 6.86 (dd, 1H), 6.78 (dd, 1H), 3.99 (s, 3H), 3.74 (br s, 2H), 2.94 (sept, 1H, 1.29 (d, 6H).
Intermediate A5: d-(2-Amino-5-fluoro-3-isopropylphenyl)picolinonitrile
[0678] ##STR00064##
[0679] 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)picolinonitrile (1 g, 4.35 mmol), 2-bromo-4-fluoro-6-isopropylaniline (Intermediate A3, Step A) (1 g, 4.31 mmol) and potassium carbonate (2 g, 14.47 mmol) were suspended in a mixture of dioxane (10 mL) and water (1 mL). After degassing with nitrogen for 15 minutes, Pd(dppf)Cl.sub.2.DCM (150 mg, 0.184 mmol) was added and the mixture was heated to 90° C. overnight, after which time complete consumption of starting bromide was seen. The mixture was cooled to room temperature and diluted with EtOAc (10 mL) and water (5 mL). The organic phase was separated, dried (MgSO.sub.4), filtered and concentrated in vacuo to give a brown oil. The crude product was purified by chromatography (Companion apparatus, 40 g column, 0-5% MeOH/DCM) to afford the product as a pale brown oil. The bulk material was further purified by SCX. It was dissolved in methanol (10 mL) and SCX (0.84 meq, 8 g, 3 eq) was added. It was stirred overnight, filtered and washed first with MeOH (100 mL) and then 0.7 M NH.sub.3 in methanol (100 mL). The ammoniacal fractions were concentrated in vacuo to afford the title compound (0.484 g, 42%) as a pale yellow oil.
[0680] .sup.1H NMR (CDCl.sub.3) δ 8.79 (dd, J=5.1, 0.9 Hz, 1H), 7.87 (dd, J=1.7, 0.9 Hz, 1H), 7.67 (dd, J=5.1, 1.7 Hz, 1H), 7.01 (dd, J=9.8, 2.9 Hz, 1H), 6.71 (dd, J=8.4, 2.9 Hz, 1H), 3.62 (br s, 2H), 2.95 (sept, J=6.8 Hz, 1H) and 1.32 (d, J=6.8 Hz, 6H).
[0681] m/z 256.4 (M+H).sup.+ (ES.sup.+).
PREPARATION OF EXAMPLES
Example 1: (R)—N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide
[0682] ##STR00065##
[0683] To a solution of (R)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Intermediate P1) (120 mg, 0.573 mmol) in THF (2 mL) was added sodium tert-butoxide (2M in THF) (0.3 mL, 0.600 mmol) and the mixture was stirred at room temperature for 1 hour. 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) (120 mg, 0.602 mmol) in THF (2 mL) was added and the mixture was stirred at room temperature overnight. The volatiles were evaporated, the residue dissolved in DMSO and purified by prep-HPLC to afford the title compound (29 mg, 12%) as a white solid.
[0684] .sup.1H NMR (DMSO-d.sub.6) δ 8.02 (s, 1H), 7.79 (d, J=1.3 Hz, 1H), 7.76 (d, J=1.3 Hz, 1H), 6.90 (s, 1H), 5.01 (d, J=4.1 Hz, 1H), 4.05-3.96 (m, 1H), 3.92-3.82 (m, 2H), 2.77 (t, J=7.4 Hz, 4H), 2.59 (t, J=7.4 Hz, 4H), 1.93 (p, J=7.4 Hz, 4H), 1.01 (d, J=5.8 Hz, 3H). One exchangeable proton not visible.
[0685] LCMS; m/z 405.3 (M+H).sup.+ (ES.sup.+).
Example 2: (S)—N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide
[0686] ##STR00066##
[0687] Prepared according to the general procedure of (R)—N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Example 1) from (S)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Intermediate P2) and 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) to afford the title compound (13 mg, 13%) as a white solid.
[0688] .sup.1H NMR (DMSO-d.sub.6) δ 8.75 (br s, 1H), 7.98 (s, 1H), 7.73 (s, 2H), 6.88 (s, 1H), 5.01 (d, J=4.1 Hz, 1H), 4.08-3.93 (m, 1H), 3.92-3.79 (m, 2H), 2.77 (t, J=7.4 Hz, 4H), 2.60 (t, J=7.4 Hz, 4H), 1.93 (p, J=7.4 Hz, 4H), 1.01 (d, J=5.8 Hz, 3H). LCMS; m/z 405.3 (M+H).sup.+ (ES.sup.+).
Example 3: 1-(2-(Dimethylamino)ethyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1H-imidazole-4-sulfonamide
[0689] ##STR00067##
[0690] To a solution of 1-(2-(dimethylamino)ethyl)-1H-imidazole-4-sulfonamide (Intermediate P3) (105 mg, 0.481 mmol) in THF (2.5 mL) was added sodium tert-butoxide (2M in THF) (0.3 mL, 0.600 mmol) and the reaction mixture was stirred at room temperature for 1 hour. 4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) (105 mg, 0.527 mmol) in THF (2.5 mL) was added and the reaction mixture was stirred at room temperature overnight. The volatiles were removed in vacuo, the residue dissolved in DMSO (2 mL) and purified by prep-HPLC to afford the title compound (44 mg, 22%) as a white solid.
[0691] .sup.1H NMR (DMSO-d.sub.6) δ 8.06 (s, 1H), 7.93 (d, J=1.3 Hz, 1H), 7.86 (d, J=1.3 Hz, 1H), 6.92 (s, 1H), 4.11 (t, J=6.2 Hz, 2H), 2.78 (t, J=7.4 Hz, 4H), 2.63-2.55 (m, 6H), 2.17 (s, 6H), 1.94 (p, J=7.4 Hz, 4H). One exchangeable proton not visible.
[0692] LCMS; m/z 418.4 (M+H).sup.+ (ES.sup.+).
Example 4: N-((2-(2-Cyanopyridin-4-yl)-4-fluoro-6-isopropylphenyl) carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide
[0693] ##STR00068##
[0694] 4-(2-Amino-5-fluoro-3-isopropylphenyl)picolinonitrile (Intermediate A5) (38 mg, 0.149 mmol) was added to (4-(dimethylamino)pyridin-1-ium-1-carbonyl)((1-isopropyl-1H-imidazol-4-yl)sulfonyl)amide (Intermediate P4) (50 mg, 0.148 mmol) in MeCN (1 mL) and the mixture was stirred at 50° C. for 2 hours. The crude product was purified by reversed phase prep-HPLC (General Methods, basic prep) to afford the title compound (19 mg, 27%) as a white solid.
[0695] .sup.1H NMR (DMSO-d.sub.6) δ 10.78 (bs, 1H), 8.68 (d, J=5.1 Hz, 1H), 8.02 (s, 2H), 7.89 (s, 1H), 7.82 (s, 1H), 7.63 (d, J=5.0 Hz, 1H), 7.28 (dd, J=10.1, 3.0 Hz, 1H), 7.16 (dd, J=8.8, 3.0 Hz, 1H), 4.46 (sept, J=6.9 Hz, 1H), 3.13-3.01 (m, 1H), 1.41 (d, J=6.7 Hz, 6H), 1.10 (d, J=6.2 Hz, 6H).
[0696] LCMS m/z 471.2 (M+H)+ (ES+).
Example 5: N-((4-Fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)phenyl) carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide
[0697] ##STR00069##
[0698] Prepared according to the general procedure of N-((2-(2-cyanopyridin-4-yl)-4-fluoro-6-isopropylphenyl)carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide (Example 4) from 4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)aniline (Intermediate A3) (39 mg, 0.150 mmol) and (4-(dimethylamino)pyridin-1-ium-1-carbonyl)((1-isopropyl-1H-imidazol-4-yl)sulfonyl)amide (Intermediate P4) (50 mg, 0.148 mmol) to afford the title compound (20 mg, 28%) as a colourless solid.
[0699] .sup.1H NMR (DMSO-d.sub.6) δ 10.55 (bs, 1H), 8.09 (d, J=5.3 Hz, 1H), 7.95 (s, 1H), 7.90 (s, 1H), 7.80 (s, 1H), 7.21 (dd, J=10.0, 3.0 Hz, 1H), 7.03 (dd, J=8.9, 3.0 Hz, 1H), 6.83 (d, J=5.3 Hz, 1H), 6.74 (s, 1H), 4.48 (sept, J=6.1 Hz, 1H), 3.88 (s, 3H), 3.02-2.93 (m, 1H), 1.41 (d, J=6.7 Hz, 6H), 1.16-0.95 (m, 6H).
[0700] LCMS m/z 476.6 (M+H)+ (ES+).
Example 6: 1-Ethyl-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-methyl-1H-imidazole-4-sulfonamide, potassium salt
[0701] ##STR00070##
[0702] Prepared as described for N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt (Example 13) using 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) and 1-ethyl-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P10) to afford the title compound (70%) as a white solid.
[0703] .sup.1H NMR (Methanol-d4) δ 7.45 (s, 1H), 6.85 (s, 1H), 3.96 (q, 2H), 2.76 (m, 8H), 2.36 (s, 3H), 1.99 (m, 4H), 1.37 (t, 3H).
[0704] LCMS m/z 389 (M+H)+ (ES+); 387 (M−H)− (ES−).
Example 7: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-isopropyl-2-methyl-1H-imidazole-4-sulfonamide, potassium salt
[0705] ##STR00071##
[0706] Prepared as described for N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt (Example 13) using 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) and 1-isopropyl-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P9) to afford the title compound (47%) as a white solid.
[0707] .sup.1H NMR (Methanol-d4) δ 7.54 (s, 1H), 6.84 (s, 1H), 4.50-4.27 (m, 1H), 2.76 (m, 8H), 2.37 (s, 3H), 1.98 (m, 4H), 1.42 (d, 6H).
[0708] LCMS m/z 403 (M+H)+ (ES+); 401 (M−H)− (ES−).
Example 8: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-5-sulfonamide, potassium salt
[0709] ##STR00072##
[0710] Prepared as described N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt (Example 13) using 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) and 2-isopropyl-1-methyl-1H-imidazole-5-sulfonamide (Intermediate P7) to afford the title compound (57%) as a white solid.
[0711] .sup.1H NMR (Methanol-d4) δ 7.30 (s, 1H), 6.85 (s, 1H), 3.86 (s, 3H), 3.19-3.04 (m, 1H), 2.91-2.52 (m, 8H), 2.10-1.83 (m, 4H), 1.29 (d, 6H).
[0712] LCMS m/z 403 (M+H)+ (ES+); 401 (M−H)− (ES−).
Example Q: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-methyl-1H-imidazole-2-sulfonamide, potassium salt
[0713] ##STR00073##
[0714] Prepared as described for N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt (Example 13) using 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) and 1-methyl-1H-imidazole-2-sulfonamide to afford the title compound (62%) as a white solid.
[0715] .sup.1H NMR (Methanol-d4) δ 7.13 (s, 1H), 6.93 (s, 1H), 6.85 (s, 1H), 3.96 (s, 3H), 2.73 (m, 8H), 1.97 (m, 4H).
[0716] LCMS m/z 361 (M+H)+ (ES+); 359 (M−H)− (ES−).
Example 10: N-((4-Fluoro-2-isopropyl-6-(i-methyl-1H-pyrazol-4-yl) phenyl)carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide
[0717] ##STR00074##
[0718] Prepared according to the general procedure of N-((2-(2-cyanopyridin-4-yl)-4-fluoro-6-isopropylphenyl)carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide (Example 4) from 4-fluoro-2-isopropyl-6-(2-methoxypyridin-4-yl)aniline (Intermediate A4) (34.6 mg, 0.148 mmol) and (4-(dimethylamino)pyridin-1-ium-1-carbonyl)((1-isopropyl-1H-imidazol-4-yl)sulfonyl)amide (Intermediate P4) (50 mg, 0.148 mmol) to afford the title compound (24.9 mg, 37%) as a colourless solid.
[0719] .sup.1H NMR (DMSO-d.sub.6) δ 7.95 (s, 1H), 7.90 (s, 1H), 7.81 (s, 1H), 7.68 (s, 1H), 7.68-7.64 (m, 1H), 7.14 (dd, J=9.9, 3.0 Hz, 1H), 6.94 (dd, J=10.0, 3.0 Hz, 1H), 4.44 (sept, J=6.3 Hz, 1H), 3.87 (s, 3H), 3.14-2.87 (m, 1H), 1.38 (d, J=6.7 Hz, 6H), 1.04 (d, J=6.8 Hz, 6H). NH not observed.
[0720] LCMS m/z 449.4 (M+H)+ (ES+).
Example 11: N-((1,2,3,5,6,7-Hexahydro-s-indacen-,4-yl)carbamoyl)-1,2-dimethyl-1H-imidazole-4-sulfonamide
[0721] ##STR00075##
[0722] Prepared according to the general procedure of (R)—N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Example 1) from 1,2-dimethyl-1H-imidazole-4-sulfonamide and 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) to afford the title compound (14 mg, 15%) as a white solid.
[0723] .sup.1H NMR (DMSO-d.sub.6) δ 10.48 (br s, 1H), 8.10 (s, 1H), 7.82 (s, 1H), 6.94 (s, 1H), 3.35 (s, 3H), 2.79 (t, J=7.4 Hz, 4H), 2.59 (t, J=7.3 Hz, 4H), 2.32 (s, 3H), 1.99-1.92 (m, 4H).
[0724] LCMS m/z 375 (M+H).sup.+ (ES.sup.+).
Example 12: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-methyl-1H-imidazole-4-sulfonamide
[0725] ##STR00076##
[0726] Prepared according to the general procedure of (R)—N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Example 1) from 1-methyl-1H-imidazole-4-sulfonamide and 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) to afford the title compound (21 mg, 24%) as a white solid.
[0727] .sup.1H NMR (DMSO-d.sub.6) δ 10.58 (br s, 1H), 8.05 (s, 1H), 7.89 (s, 1H), 7.83 (s, 1H), 6.93 (s, 1H), 3.70 (s, 3H), 2.78 (t, J=7.4 Hz, 4H), 2.58 (t, J=7.4 Hz, 4H), 1.97-1.90 (m, 4H).
[0728] LCMS m/z 361 (M+H).sup.+ (ES.sup.+).
Example 13: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt
[0729] ##STR00077##
[0730] To a solution of 2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide (Intermediate P8) (85 mg, 0.42 mmol) in THF (3 mL) was added potassium tert-butoxide (47 mg, 0.42 mmol). The mixture was stirred for 40 minutes. A solution of 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) (83 mg, 0.41 mmol) in THF (1 mL) was added and the mixture was stirred overnight at room temperature. The reaction mixture was concentrated in vacuo and DMSO (1 mL) was added. The solution (suspensions were filtered first over cotton wool) was submitted for purification by reversed phase column chromatography (see “Experimental Methods”) to afford the title compound (44 mg, 26%) as a white solid.
[0731] .sup.1H NMR (Methanol-d4) δ 7.36 (s, 1H), 6.85 (s, 1H), 3.64 (s, 3H), 3.17-3.01 (m, 1H), 2.75 (m, 8H), 1.99 (m, 4H), 1.29 (d, 6H).
[0732] LCMS m/z 403 (M+H)+ (ES+); 401 (M−H)− (ES−).
Example 14: N-((2,6-Diisopropylphenyl)carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide, sodium salt
[0733] ##STR00078##
[0734] Prepared according to the general procedure of (R)—N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxypropyl)-1H-imidazole-4-sulfonamide (Example 1) from 1-isopropyl-1H-imidazole-4-sulfonamide and 2-isocyanato-1,3-diisopropylbenzene (Intermediate A2) to afford the title compound (27 mg, 28%) as a colourless solid.
[0735] .sup.1H NMR (DMSO-d.sub.6) δ 7.65 (d, J=1.4 Hz, 1H), 7.45 (br s, 2H), 7.13-7.05 (m, 1H), 7.01 (d, J=7.6 Hz, 2H), 4.44-4.38 (m, 1H), 3.15-3.07 (m, 2H), 1.38 (d, J=6.7 Hz, 6H), and 1.03 (d, J=6.9 Hz, 12H).
[0736] LCMS m/z 393 (M+H).sup.+ (ES.sup.+).
Example 15: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-isopropyl-1H-imidazole-4-sulfonamide, sodium salt
[0737] ##STR00079##
[0738] 1-Isopropyl-1H-imidazole-4-sulfonamide (35.5 mg, 0.188 mmol) was dissolved in THF (5 mL) and 2 M sodium tert-butoxide in THF (0.098 ml, 0.197 mmol) added. After the mixture had been stirred at room temperature for 1 hour, 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) (37.4 mg, 0.188 mmol) was added and the mixture stirred at room temperature for 15 hours. Ethyl acetate (5 mL) was added to the mixture and the suspension filtered and washed with ethyl acetate (1 mL). The collected solid was dried under reduced pressure to afford the title compound (32 mg, 40%) as a white solid.
[0739] .sup.1H NMR (DMSO-d.sub.6) δ 7.67 (d, J=1.5 Hz, 1H), 7.62 (br s, 1H), 7.45 (d, J=1.5 Hz, 1H), 6.77 (s, 1H), 4.45-4.39 (m, 1H), 2.75 (t, J=7.4 Hz, 4H), 2.64 (t, J=7.3 Hz, 4H), 1.93-1.86 (m, 4H), and 1.39 (d, J=6.7 Hz, 6H).
[0740] LCMS m/z 389 (M+H).sup.+ (ES.sup.+).
Example 16: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-2-methyl-1-((tetrahydrofuran-2-yl)methyl)-1H-imidazole-4-sulfonamide, potassium salt
[0741] ##STR00080##
[0742] Prepared as described for N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt (Example 13) using 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) and 2-methyl-1-((tetrahydrofuran-2-yl)methyl)-1H-imidazole-4-sulfonamide (Intermediate P11) to afford the title compound (53%) as a white solid.
[0743] .sup.1H NMR (Methanol-d4) δ 7.49 (s, 1H), 6.85 (s, 1H), 4.18-4.01 (m, 2H), 4.01-3.86 (m, 1H), 3.88-3.59 (m, 2H), 2.76 (m, 9H), 2.38 (s, 3H), 2.18-1.91 (m, 4H), 1.89-1.71 (m, 2H), 1.70-1.41 (m, 1H).
[0744] LCMS m/z 445 (M+H)+ (ES+); 443 (M−H)− (ES−).
Example 17: N-((1,2,3,5,6,7-Hexahydro-s-indacen-4-yl)carbamoyl)-1-(2-hydroxyethyl)-2-methyl-1H-imidazole-4-sulfonamide, potassium salt
[0745] ##STR00081##
[0746] Prepared as described for N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-isopropyl-1-methyl-1H-imidazole-4-sulfonamide, potassium salt (Example 13) using 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) and 1-(2-hydroxyethyl)-2-methyl-1H-imidazole-4-sulfonamide (Intermediate P12) and one additional equivalent of KOtBu to afford the title compound (14%) as a white solid.
[0747] .sup.1H NMR (Methanol-d4) δ 7.60 (s, 1H), 6.90 (s, 1H), 4.04 (d, 2H), 3.80 (t, 2H), 2.94-2.61 (m, 9H), 2.41 (s, 3H), 2.12-1.94 (m, 4H).
[0748] LCMS m/z 405 (M+H)+ (ES+).
Example 18: 2-((Dimethylamino)methyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-methyl-1H-imidazole-4-sulfonamide, and
Example 19: 5-((Dimethylaminomethyl)-N-((1,2,2,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-methyl-1H-imidazole-4-sulfonamide
[0749] ##STR00082##
[0750] Sodium tert-butoxide, 2 M in THF (0.120 mL, 0.241 mmol) was added to a solution of a mixture of 2-((dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide and 5-((dimethylamino)methyl)-1-methyl-1H-imidazole-4-sulfonamide (Intermediates P5 and P6) (50 mg, 0.229 mmol) in THF (1 mL) and stirred at room temperature for 1 hour. Then 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (Intermediate A1) (47.9 mg, 0.241 mmol) in THF (1 mL) was added and the reaction stirred at room temperature over the weekend. The reaction mixture was concentrated and the crude product was purified by chromatography (Companion apparatus, RP Flash C18, 12 g column, 5.50% MeCN/10 mM ammonium bicarbonate) to afford 2-((dimethylamino)methyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-methyl-1H-imidazole-4-sulfonamide (Example 18) (44 mg, 46%) and 5-((dimethylamino)methyl)-N-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-1-methyl-1H-imidazole-4-sulfonamide (Example 19) (3 mg, 3%), both as colourless solids.
[0751] Example 18: .sup.1H NMR (DMSO-d.sub.6) δ 7.91 (s, 1H), 7.77 (s, 1H), 6.83 (s, 1H), 4.26 (s, 2H), 3.67 (s, 3H), 2.76 (t, J=7.4 Hz, 4H), 2.66-2.53 (m, 10H), 1.96-1.84 (m, 4H) (1 exchangeable NH not observed).
[0752] LCMS; m/z 418.3 (M+H).sup.+ (ES.sup.+).
[0753] Example 19: .sup.1H NMR (DMSO-d.sub.6) δ 10.70 (br s, 1H), 7.96 (s, 1H), 7.80 (s, 1H), 6.90 (s, 1H), 3.68 (s, 3H), 346 (s, 2H), 2.78 (t, J=7.4 Hz, 4H), 2.58 (t, J=7.4 Hz, 4H), 2.14 (s, 6H), 1.99-1.86 (m, 4H).
[0754] LCMS; m/z 418.3 (M+H).sup.+ (ES.sup.+).
[0755] The compounds of examples 20-21 were synthesised by methods analogous to those outlined above.
TABLE-US-00001 TABLE 1 1H NMR and MS data Ex Structure and Name .sup.1H NMR spectrum MS MW 20
Examples—Biological Studies
[0756] NLRP3 and Pyroptosis
[0757] 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), 62579; Alexander Wree et al., Hepatology, 2014, 59(3), 898-910; Alex Baldwin et al., Journal of Medicinal Chemistry, 2016, 59(5), 1691-1710; Ema Ozaki et al., Journal of Inflammation Research, 2015, 8, 15-27; Zhen Xie & Gang Zhao, Neuroimmunology Neuroinflammation, 2014, 1(2), 60-65; Mattia Cocco et al., Journal of Medicinal Chemistry, 2014, 57(24), 10366-10382; T. Satoh et al., Cell Death & Disease, 2013, 4, e644). Therefore, it is anticipated that inhibitors of NLRP3 will block pyroptosis, as well as the release of pro-inflammatory cytokines (e.g. IL-1β) from the cell.
[0758] THP-1 Cells: Culture and Preparation
[0759] THP-1 cells (ATCC #TIB-202) were grown in RPMI containing L-glutamine (Gibco #11835) supplemented with 1 mM sodium pyruvate (Sigma #S8636) and penicillin (100 units/ml)/streptomycin (0.1 mg/ml) (Sigma #P4333) in 10% Fetal Bovine Serum (FBS) (Sigma #F0804). The cells were routinely passaged and grown to confluency (˜10.sup.6 cells/ml). On the day of the experiment, THP-1 cells were harvested and resuspended into RPMI medium (without FBS). The cells were then counted and viability (>90%) checked by Trypan blue (Sigma #T8154). Appropriate dilutions were made to give a concentration of 625,000 cells/ml. To this diluted cell solution was added LPS (Sigma #L4524) to give a 1 μg/ml Final Assay Concentration (FAC). 40 μl of the final preparation was aliquoted into each well of a 96-well plate. The plate thus prepared was used for compound screening.
[0760] THP-1 Cells Pyroptosis Assay
[0761] The following method step-by-step assay was followed for compound screening. [0762] 1. Seed THP-1 cells (25,000 cells/well) containing 1.0 μg/ml LPS in 40 μl of RPMI medium (without FBS) in 96-well, black walled, clear bottom cell culture plates coated with poly-D-lysine (VWR #734-0317) [0763] 2. Add 5 μl compound (8 points half-log dilution, with 10 μM top dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells [0764] 3. Incubate for 3 hrs at 37° C. in 5% CO.sub.2 [0765] 4. Add 5 μl nigericin (Sigma #N7143) (FAC 5 μM) to all wells [0766] 5. Incubate for 1 hr at 37° C. and 5% CO.sub.2 [0767] 6. At the end of the incubation period, spin plates at 300×g for 3 mins and remove supernatant [0768] 7. Then add 50 μl of resazurin (Sigma #R7017) (FAC 100 μM resazurin in RPMI medium without FBS) and incubate plates for a further 1-2 hrs at 37° C. and 5% CO.sub.2 [0769] 8. Plates were read in an Envision reader at Ex 560 nm and Em 590 nm [0770] 9. IC.sub.50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)
[0771] 96-Well Plate Map
TABLE-US-00002 1 2 3 4 5 6 7 8 9 10 11 12 A High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low B High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low C High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low D High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low E High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low F High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low G High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low H High Comp 1 Comp 2 Comp 3 Comp 4 Comp 5 Comp 6 Comp 7 Comp 8 Comp 9 Comp 10 Low High MCC950(10 uM) Compound 8-point half-log dilution Low Drug free control
[0772] The results of the pyroptosis assay performed are summarised in Table 2 below as THP IC.sub.50.
[0773] Human Whole Blood IL1β Release Assay
[0774] 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.
[0775] Human whole blood in Li-heparin tubes was obtained from healthy donors from a volunteer donor panel. [0776] 1. Plate out 80 μl of whole blood containing 1 μg/ml of LPS in 96-well, clear bottom cell culture plate (Corning #3585) [0777] 2. Add 10 μl compound (8 points half-log dilution with 10 μM top dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells [0778] 3. Incubate for 3 hrs at 37° C. 5% CO.sub.2 [0779] 4. Add 10 μl Nigericin (Sigma #N7143) (10 μM FAC) to all wells [0780] 5. Incubate for 1 hr at 37° C. 5% CO.sub.2 [0781] 6. At the end of the incubation period, spin plates at 300×g for 5 mins to pellet cells and remove 20 μl of supernatant and add to 96-well v-bottom plates for IL-1β analysis (note: these plates containing the supernatants can be stored at −80° C. to be analysed at a later date) [0782] 7. IL-1β was measured according to the manufacturer protocol (Perkin Elmer-AlphaLisa IL-1 Kit AL220F-5000) [0783] 8. IC.sub.50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)
[0784] The results of the human whole blood assay are summarised in Table 2 below as HWB IC.sub.50.
TABLE-US-00003 TABLE 2 NLRP.sub.3 inhibitory activity Example No THP IC.sub.50 HWB IC.sub.50 1 +++ *** 2 +++ *** 3 ++ ** 4 +++ **** 5 ++++ *** 6 +++ * 7 +++ ** 8 +++ ND 9 + ND 10 + ND 11 ++ ND 12 ++ ND 13 ++ * 14 + ND 15 +++ ** 16 +++ *** 17 ++ *** 18 + ND 19 +++ *** 20 +++++ ***** 21 ++++ **** [THP IC.sub.50 (≤0.04 μM = +++++, ≤0.16 μM = ++++, ≤0.64 μM = +++, ≤2.56 μM = ++, ≤10 μM = +, not determined = ND)] [HWB IC.sub.50 (≤0.4 μM = *****, ≤0.8 μM = ****, ≤1.6 μM = ***, ≤3.2 μM = **, ≤10 μM = *, not determined = ND)]
[0785] PK Protocol
[0786] Pharmacokinetic parameters were determined in male Sprague Dawley rats (Vital River Laboratory Animal Technology Co Ltd, Beijing, China, 7-9 weeks old). Animals were individually housed during the study and maintained under a 12 h light/dark cycle. Animals had free access to food and water.
[0787] For intravenous administration, compounds were formulated as a solution in DMSO:PBS [10:90] in 2 mL/kg dosing volume and administered via tail vein.
[0788] Serial blood samples (about 200 μL) were taken from each animal at each of 8 time-points post dose (0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 h). Samples were held on ice for no longer than 30 minutes before centrifugation (5,696 rpm (3,000 g) for 15 minutes) for plasma generation. Plasma was frozen on dry ice prior to bioanalysis. PK parameters were generated from LC-MS/MS data using Phoenix WinNonlin 6.3 software.
TABLE-US-00004 TABLE 3 PK data (intravenous administration) Example Dose AUC T.sub.1/2 V.sub.dss Cl No (mg/kg) (ng .Math. hr/mL) (hr) (L/kg) (mL/min/kg) 4 1 200 1.6 2.5 85.4
[0789] As is evident from the results presented in Table 2, surprisingly in spite of the structural differences versus the prior art compounds, the compounds of the invention show high levels of NLRP3 inhibitory activity in the pyroptosis assay and in the human whole blood assay.
[0790] As is evident from the results presented in Table 3, the compounds of the invention show advantageous pharmacokinetic properties, for example half-life T.sub.1/2, area under the curve AUC, clearance Cl and/or bioavailability, compared to the prior art compounds.
[0791] 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.