Kynurenine-3-monooxygenase inhibitors, pharmaceutical compositions, and methods of use thereof

Abstract

Certain chemical entities are provided herein. Also provided are pharmaceutical compositions comprising at least one chemical entity and one or more pharmaceutically acceptable vehicle. Methods of treating patients suffering from certain diseases and disorders responsive to the inhibition of KMO activity are described, which comprise administering to such patients an amount of at least one chemical entity effective to reduce signs or symptoms of the disease or disorder are disclosed. These diseases include neurodegenerative disorders such as Huntington's disease. Also described are methods of treatment include administering at least one chemical entity as a single active agent or administering at least one chemical entity in combination with one or more other therapeutic agents. Also provided are methods for screening compounds capable of inhibiting KMO activity.

Claims

1. A compound of formula: ##STR00164## or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition comprising the compound of claim 1 and at least one pharmaceutically acceptable excipient.

3. A method of treating Huntington's disease in a subject in need of such a treatment which method comprises administering to the subject a therapeutically effective amount of the compound of claim 1.

Description

EXAMPLES

(1) The chemical entities, compositions, and methods described herein are further illustrated by the following non-limiting examples.

(2) As used herein, the following abbreviations have the following meanings. If an abbreviation is not defined, it has its generally accepted meaning CDI=carbonyldiimidazole DCM=dichloromethane DME=dimethyl ether DMEM=Dulbecco's modified Eagle's medium DMF=N,N-dimethylformamide DMSO=dimethylsulfoxide EDC.HCl=1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride EtOH=ethanol Et.sub.2O=diethylether EtOAc=ethyl acetate g=gram hr=hour hrs=hours HOBt=1-Hydroxybenzotriazol LiHMDS=lithium hexamethyl-disilazide LC/MS=liquid chromatography/mass spectrometry mg=milligram min=minutes mL=milliliter mmol=millimoles mM=millimolar ng=nanogram nm=nanometer nM=nanomolar PBS=phosphate buffered saline rt=room temperature TBME=t-butyl methyl ether THF=tetrahydrofuran TMOF=trimethylorthoformate ?L=microliter ?M=micromolar 1 g/1 ml=1 vol

Experimental

(3) Commercially available reagents and solvents (HPLC grade) were used without further purification.

(4) Thin-layer chromatography (TLC) analysis was performed with Kieselgel 60 F254 (Merck) plates and visualized using UV light. Microwave reactions were carried out using CEM focussed microwaves.

(5) Analytical HPLC-MS was performed on Agilent HP1100 and Shimadzu 2010, systems using reverse phase Atlantis dC18 columns (5 ?m, 2.1?50 mm), gradient 5-100% B (A=water/0.1% formic acid, B=acetonitrile/0.1% formic acid) over 3 min, injection volume 3 ?l, flow=1.0 ml/min. UV spectra were recorded at 215 nm using a Waters 2487 dual wavelength UV detector or the Shimadzu 2010 system. Mass spectra were obtained over the range m/z 150 to 850 at a sampling rate of 2 scans per second using Waters ZMD and over m/z 100 to 1000 at a sampling rate of 2 Hz using Electrospray ionisation, by a Shimadzu 2010 LC-MS system or analytical HPLC-MS was performed on Agilent HP1100 and Shimadzu 2010, systems using reverse phase Water Atlantis dC18 columns (3 ?m, 2.1?100 mm), gradient 5-100% B (A=water/0.1% formic acid, B=acetonitrile/0.1% formic acid) over 7 min, injection volume 3 ?l, flow=0.6 ml/min. UV spectra were recorded at 215 nm using a Waters 2996 photo diode array or on the Shimadzu 2010 system. Mass spectra were obtained over the range m/z 150 to 850 at a sampling rate of 2 scans per second using Waters ZQ and over m/z 100 to 1000 at a sampling rate of 2 Hz using Electrospray ionisation, by a Shimadzu 2010 LC-MS system. Data were integrated and reported using OpenLynx and OpenLynx Browser software or via Shimadzu PsiPort software.

Example 1

(6) ##STR00006##

(7) Referring to Reaction Scheme 1, Stage 1, to a stirred suspension of dichloropyrimidine (1 eq) in 1,4-dioxane (15 vol) was added boronic acid (0.7 eq) and Pd(PPh3)4 (0.025 eq). A 2M K2CO3 solution (7.5 vol) was added to the resulting mixture, which was heated at 90? C. overnight under an atmosphere of N2. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was dissolved in EtOAc:water (1:1) (100 vol) and the resulting solution filtered through celite. The organic layer was separated and the aqueous layer further extracted with EtOAc (50 vol). The combined organic layers were washed with saturated aqueous NaCl (20 vol), dried over Na2SO4, filtered and the solvent removed in vacuo. The resulting residue was purified by flash column chromatography (eluent: [0:1 to 1:19] EtOAc:heptane) to afford the required target compounds.

(8) Referring to Reaction Scheme 1, Stage 2, 4-chloro-6-substituted-phenyl-pyrimidine (1 eq), PdCl2(dppf).DCM (0.05 eq) and triethylamine (2 eq) were suspended in degassed MeOH (50 vol) in a bomb fitted with a magnetic stirrer bar. The atmosphere in the reaction vessel was replaced with N2 by successive evacuation and charging with N2 gas (this process was repeated three times). The bomb was then flushed with CO by successive charging with CO and evacuation. The vessel was pressurised to 5 bar of CO and heated at 50? C. with stirring for 5 hours. The reaction vessel was allowed to cool to room temperature before venting CO and flushing with N2. The reaction mixture was concentrated in vacuo and the resulting residue dissolved in EtOAc (30 vol) and water (30 vol). The solution was filtered through cotton wool and the organic layer was separated, washed with saturated aqueous NaCl (15 vol), dried over Na2SO4, filtered and concentrated under reduced pressure. Purification by flash column chromatography (eluent: [0:1 to 1:9] EtOAc:heptane) yielded the target compounds.

(9) Referring to Reaction Scheme 1, Stage 3, 6-substituted-phenyl-pyrimidine-4-carboxylic acid methyl ester (1 eq) was suspended in MeOH (20 vol), 1M NaOH solution (20 vol) and stirred at room temperature for 4 hours. The reaction mixture was acidified with 2M HCl. Soluble products were extracted with DCM (2?20 vol) and the combined organic layers were dried over MgSO4, filtered and concentration under reduced pressure afforded the target compounds. Insoluble products were filtered, washed with water (3?10 vol) and heptane (3?10 vol) before drying in vacuo to yield the target compounds.

(10) Referring to Reaction Scheme 1, Stage 4, the required amide analogues were prepared following the procedures described in method A, B, C or D.

(11) The following compounds were prepared substantially as described above.

(12) TABLE-US-00001 Molecular Structure Weight Mass Spec Result 264.67 [M + H].sup.+ = 265/267, 100% @ rt = 3.53 and 3.70 min 276.72 [M + H].sup.+ = 277/279, 99.9% @ rt = 4.32 min 232.22 [M + H].sup.+ = 232, 100% @ rt = 3.52 min 0 246.24 [M + H].sup.+ = 247, 100% @ rt = 3.66 min 260.27 [M + H].sup.+ = 261.4, 100% @ rt = 4.13 min 251.25 [M + H].sup.+ = 252, 99% @ rt = 2.32 min 319.75 [M + H]+ = 320, 97% @ rt = 2.29 min

Example 2

(13) ##STR00014##

(14) Referring to Reaction Scheme 2, Stage 1, to a degassed stirred solution of 4-chloro-3-nitro-benzene boronic acid (1 eq) and 4,6-dichloropyrimidine (1.44 eq) in 1,4-dioxane (16 vol) and 2N K2CO3 (8 vol) was added Pd(PPh3)4 (0.06 eq) and the mixture heated to 90? C. for 3.75 hours under an atmosphere of nitrogen gas. The cooled reaction mixture had the solvents removed under reduced pressure. DCM (25 vol) and water (25 vol) were then added and the undissolved material removed by filtration through celite. The organic phase from the filtrate was concentrated under reduced pressure whilst adsorbing on to silica gel (8.2 g). The residue was purified using dry flash chromatography (gradient up to 10% EtOAc:heptane) to afford the target compound.

(15) Referring to Reaction Scheme 2, Stage 2, in a metal vessel equipped to carry out high pressure reactions, a degassed suspension of 4-chloro-6-(4-chloro-3-nitro-phenyl)-pyrimidine (1 eq) was stirred in MeOH (62 vol). Triethylamine (2 eq) and Pd(PPh3)4 (0.05 eq) was then added and the vessel sealed. The vessel was then charged with carbon monoxide gas to a pressure of 5 bar and heated to 50? C. for 18 hours. After extrusion of excess carbon monoxide gas, the organic solvent was concentrated under reduced pressure. To the residue was added DCM (26 vol) and the undissolved material was filtered off and washed with DCM (10 vol). The filtrate was washed with 2N HCl (10 vol), a 1:1 mixture of water and brine (10 vol) and then concentrated under reduce pressure whilst adsorbing onto silica gel (3.2 g). The residue was purified by dry flash column chromatography (gradient up to 60% EtOAc:heptane) to give a mixture of products, the major identified as the methyl ester. The solid was then dissolved in 2N HCl (30 vol) and washed with TBME (1?30 vol & 1?20 vol). The aqueous layer was adjusted to pH7 and the precipitate formed was filtered off, washed with water (2?5 vol) and air dried to afford the target compound.

(16) TABLE-US-00002 Molecular Mass Spec Structure Weight Result 249.66 [M + H].sup.+ = 250/252, 96% @ rt = 3.43 min

Example 3

(17) ##STR00016## ##STR00017##

(18) Referring to Reaction Scheme 3, Stage 1, 5-bromo-2-chloro anisole (1 eq) in toluene (8 vol) and THF (3 vol) at ?78? C. was added n-BuLi (1.5 eq) drop wise. The resulting mixture was stirred at ?78? C. for 30 minutes under an atmosphere of N2. Trimethylborate (2 eq) was then added to the reaction mixture and this was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was quenched with 1M HCl and the organic layer was separated. The organic layer was washed with saturated aqueous NaCl (20 vol), dried over Na2SO4, filtered and the solvent removed in vacuum. The resulting residue was purified by flash column chromatography (eluent: [1:1] EtOAc:heptane) to afford the required target compound (1.15 g, 31%).

(19) Referring to Reaction Scheme 3, Stage 2, to a stirred suspension of dichloropyrimidine (1 eq) in 1,4-dioxane (20 vol) was added boronic acid (0.7 eq) and Pd(PPh3)4 (0.05 eq). A 2M K2CO3 solution (10 vol) was added to the resulting mixture, which was heated at 90? C. for 3 hours under an atmosphere of N2. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was dissolved in EtOAc water (1:1) (100 vol) and the resulting solution filtered through celite. The organic layer was separated and the aqueous layer further extracted with EtOAc (50 vol). The combined organic layers were washed with saturated aqueous NaCl (20 vol), dried over Na2SO4, filtered and the solvent removed in vacuo. The resulting residue was purified by flash column chromatography (eluent: [1:8] EtOAc:heptane) to afford the required target compound (1.14 g, 73%).

(20) Referring to Reaction Scheme 3, Stage 3, 4-chloro-6-substituted-phenyl-pyrimidine (1 eq), PdCl2(dppf).DCM (0.05 eq) and triethylamine (2 eq) were suspended in degassed MeOH (50 vol) in a bomb fitted with a magnetic stirrer bar. The atmosphere in the reaction vessel was replaced with N2 by successive evacuation and charging with N2 gas (this process was repeated three times). The bomb was then flushed with CO by successive charging with CO and evacuation. The vessel was pressurised to 5 bar of CO and heated at 50? C. with stirring for 16 hours. The reaction vessel was allowed to cool to room temperature before venting CO and flushing with N2. The reaction mixture was concentrated in vacuo and the resulting residue dissolved in EtOAc (30 vol) and water (30 vol). The organic layer was separated, washed with saturated aqueous NaCl (15 vol), dried over Na2SO4, filtered and concentrated under reduced pressure. Purification by flash column chromatography (eluent: [2:3] EtOAc:heptane) yielded the target compound (1.15 g, 96%).

(21) Referring to Reaction Scheme 3, Stage 4, to a solution of 6-Substituted-phenyl-pyrimidine-4-carboxylic acid methyl ester (1 eq) in DCM (80 vol) at ?78? C. was added BBr3 (3 eq) under nitrogen. The reaction mixture was warm to 0? C. and stirred for 1 hour then allowed to stir at room temperature for 16 hours. The reaction mixture was poured into ice (100 vol) and extracted with EtOAc (150 vol). The organic layer was separated, washed with saturated aqueous NaCl (15 vol), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude mixture (0.45 g) was used in the next step without further purification.

(22) Referring to Reaction Scheme 3, Stage 5, a solution of 6-substituted-phenyl-pyrimidine-4-carboxylic acid (1 eq) in MeOH (100 vol) was added concentrated H2SO4 (2 drops). The reaction mixture was refluxed for 4 hours. The reaction mixture was concentrated in vacuo and the resulting residue dissolved in EtOAc (30 vol) and water (30 vol). The organic layer was separated, washed with saturated aqueous NaCl (15 vol), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude mixture (0.48 g) was used in the next step without further purification.

(23) Referring to Reaction Scheme 3, Stage 6, to a solution of 6-substituted-phenyl-pyrimidine-4-carboxylic acid methyl ester (1.05 eq) in THF (10 vol) were added 3-hydroxy furan (1 eq) and PPh3 (1.5 eq) under nitrogen. The reaction mixture was cooled to 0? C. and DIAD (1.5 eq) was added slowly. Reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was concentrated in vacuo and the resulting residue was triturated with EtOAc and heptane (1:2) and solid was filtered to give the desired compound (0.42 g, 70%).

(24) Referring to Reaction Scheme 3, Stage 7, 6-substituted-phenyl-pyrimidine-4-carboxylic acid methyl ester (1 eq) was suspended in THF (20 vol), 2M NaOH (3.14 ml, 6.28 mmol, 5 eq) and stirred at room temperature for 4 hours. The THF was removed under vacuo, MeCN (10 vol) was added and the reaction mixture was acidified with 6M HCl. The resulting solid was filtered and washed with water and a mixture of MeCN: water (1:1) to give desired product (0.335 g. 83%).

(25) Referring to Reaction Scheme 3, Stage 3, the required amide analogue was prepared following the procedure described in method B.

(26) The following compounds were prepared substantially as described above.

(27) TABLE-US-00003 Molecular Structure Weight Mass Spec Result 320.73 [M + H].sup.+ = 321/323, 100% @ rt = 3.55-3.82 min 396.84 [M + H].sup.+ = 397/399, 98% @ rt = 3.7 min

Example 4

(28) ##STR00020## ##STR00021##

(29) Referring to Reaction Scheme 4, Stage 1, N-(2-hydroxyethyl)morpholine (1 eq) in DCM (70 vol) at 0? C. was added dibromo triphenyl phosphorane (1.2 eq). The reaction mixture was allowed to warm to room temperature and stirred for 16 hrs. The solvent removed in vacuum. DCM (10 vol) was added to the reaction mixture. The precipitate was filtered to afford the target compound. The crude mixture was used in the next step without further purification.

(30) Referring to Reaction Scheme 4, Stage 2, N-(2-bromoethyl)morpholine (1.1 eq) in DMF (15 vol) were added 2-chloro-5-iodophenol (1 eq) and Cs2CO3 (2.5 eq). The reaction mixture was refluxed for 3 hours under nitrogen. The reaction mixture was allowed to cool to room temperature and EtOAc (40 vol) and aq ammonia (40 vol) were added. The organic layer was separated and the aqueous layer further extracted with EtOAc (50 vol). The combined organic layers were washed with saturated aqueous NaCl (20 vol), dried over Na2CO3, filtered and the solvent removed in vacuo. The resulting residue was purified by flash column chromatography (eluent: [3:1] EtOAc:heptane) to afford the required target compound.

(31) Referring to Reaction Scheme 4, Stage 3, to a stirred suspension of 3-substituted-4-chloro-iodobenzene (1 eq) in degassed DMF (15 vol) was added bis-diborane (1.05 eq), Pd(OAc)2 (0.04 eq) and KOAc (3.0 eq). The reaction mixture was heated at 90? C. for 5 hrs under an atmosphere of N2. The reaction mixture was cooled to room temperature and filtered through celite then concentrated in vacuo to give crude product. Crude was used in the next step without further purification.

(32) Referring to Reaction Scheme 4, Stage 4, to a stirred suspension of dichloropyrimidine (1 eq) in 1,4-dioxane (90 vol) was added boronic ester (1.0 eq) and Pd(PPh3)4 (0.03 eq). A 2M K2CO3 (3 eq) solution was added to the resulting mixture, which was heated at 90? C. for 16 hrs under an atmosphere of N2. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was dissolved in EtOAc: water (1:1) (100 vol) and the resulting solution filtered through celite. The organic layer was separated and the aqueous layer further extracted with EtOAc (50 vol). The combined organic layers were washed with saturated aqueous NaCl (20 vol), dried over Na2SO4, filtered and the solvent removed in vacuo. The resulting residue was purified by flash column chromatography (eluent: [3:1] EtOAc:heptane) to afford the required target compound.

(33) Referring to Reaction Scheme 4, Stage 5, 4-chloro-6-substituted-phenyl-pyrimidine (1 eq), PdCl2(dppf).DCM (0.05 eq) and triethylamine (2 eq) were suspended in degassed MeOH (50 vol) in a bomb fitted with a magnetic stirrer bar. The atmosphere in the reaction vessel was replaced with N2 by successive evacuation and charging with N2 gas (this process was repeated three times). The bomb was then flushed with CO by successive charging with CO and evacuation. The vessel was pressurised to 5 bar of CO and heated at 50? C. with stirring for 16 hours. The reaction vessel was allowed to cool to room temperature before venting CO and flushing with N2. The reaction mixture was concentrated in vacuo and the resulting residue dissolved in EtOAc (30 vol) and water (30 vol). The organic layer was separated, washed with saturated aqueous NaCl (15 vol), dried over Na2SO4, filtered and concentrated under reduced pressure. Purification by re-crystallisation using MeOH yielded the target compound.

(34) Referring to Reaction Scheme 4, Stage 6, 6-substituted-phenyl-pyrimidine-4-carboxylic acid methyl ester (1 eq) was suspended in THF (20 vol), 2M NaOH (2.5 eq) and stirred at room temperature for 4 hours. Solvent (THF) was removed and reaction mixture was acidified with 2M HCl. Resulting solid was filtered and was with water to give desired product. Referring to Reaction Scheme 4, Stage 7, the required amide analogue was prepared following the procedure described in method B.

(35) The following compounds were prepared substantially as described above.

(36) TABLE-US-00004 Structure Molecular Weight Mass Spec Result 400.26 [M + H].sup.+ = 364, 98% @ rt = 2.41 min 439.91 [M + H].sup.+ = 440, 99% @ rt = 2.54 min 292.72 [M + H].sup.+ = 293/295, 100% @ rt = 4.18 min 306.75 [M + H].sup.+ = 307/309, 100% @ rt = 4.10 min 318.76 [M + H].sup.+ = 319, 100% @ rt = 4.61 min 306.75 [M + H].sup.+ = 307/309, 100% @ rt = 4.37 min 306.75 [M + H].sup.+ = 307/309, 100% @ rt = 4.37 min 290.71 [M + H].sup.+ = 291/293, 100% @ rt = 3.93 min 0 346.69 [M + H].sup.+ = 347/349, 92/8% @ rt = 4.22 304.79 [M + H].sup.+ = 305/307, 100% @ rt = 4.20 min 360.82 [M + H].sup.+ = 362/364, 100% @ rt = 2.55 min 303.75 [M + H]+ = 304, 100% @ rt = 3.78 min 307.67 [M + H]+ = 286/288, 99% @ rt = 3.26 min 363.8 [M + H]+ = 364/366 100% @ rt = 2.29 min 302.72 [M ? Na]? = 303/305 100% @ rt = 4.14 min 305.89 [M + H]+ = 307/309, 95% @ rt = 3.37 min 289.71 [M + H]+ = 290, 100% @ rt = 3.74 min

Example 5

(37) ##STR00039##

(38) Referring to Reaction Scheme 5, Stage 1, 4-(chloro-6-substituted)-phenyl-pyrimidine (1 eq) was suspended in 1,4-dioxane (3 vol) and ammonium hydroxide (6 vol) was added to the suspension. The reaction mixture was heated at 95? C. in a pressure tube for 16 hours with stirring. The reaction mixture was cooled to room temperature and the precipitate was filtered off and washed with water to yield the target compound.

(39) Referring to Reaction Scheme 5, Stage 2, 6-(substituted-phenyl)-pyrimidin-4-ylamine (1 eq) was suspended in 1,4-dioxane (20 vol). Sodium hydride (6 eq) was added and the suspension was stirred for 1 hour at ambient temperature. 3-Pyridinesulfonyl chloride or benzenesulfonyl chloride (1.2 eq) were added and the reaction mixture was stirred at 80? C. for 24 hours. In the case of pyridinesulfonyl chloride derivative, the reaction was quenched by the addition of water and the solvent was removed in vacuo. Purification by flash column chromatography (eluent: [0:1 to 1:4] MeOH:EtOAc) afforded the target compound. In the case of benzenesulfonyl chloride derivative, acetonitrile/water was added and the solid filtered off. The filtrate was concentrated in vacuo and the residue was triturated in EtOAc to furnish the sodium salt as a powder. The sodium salt was then washed with a citric acid aqueous solution followed by water and dried to furnish the desired compound.

(40) Referring to Reaction Scheme 5, Stage 3, 6-substituted-phenyl-pyrimidin-4-ylamine (1 eq) was suspended in 1,4-dioxane or DMF (20 vol). Sodium hydride (3 eq) was added and the suspension stirred for 10 to 60 minutes at room temperature. The appropriate acid chloride (1.5 eq) was added and the reaction mixture stirred at room temperature for 1 hour. The reaction was monitored by LCMS. If the reaction was not complete, sodium hydride (l eq) was added to the reaction mixture, which was then heated at 50? C. for 16 hours. Upon completion, the reaction was quenched with water. If precipitation occurred, the precipitate was filtered and purified further by flash column chromatography using an appropriate eluent, if not the desired material was extracted with EtOAc. The organic layer was washed with saturated aqueous NaCl solution, dried with MgSO4, filtered and the solvent removed in vacuo. The desired compound was further purified either by trituration or prep HPLC when required.

(41) The following compounds were prepared substantially as described above.

(42) TABLE-US-00005 Molecular Structure Weight Mass Spec Result 0 401.87 [M + H]+ = 402, 99% @ rt = 4.53 min 419.87 [M + H]+ = 420, 100% @ rt = 4.61 min 485.87 [M + H]+ = 486, 100% @ rt = 5.01 min 485.87 [M + H]+ = 487, 100% @ rt = 4.91 min 419.87 [M + H]+ = 420, 99.5% @ rt = 4.51 min 365.84 [M + H]+ = 366, 100% @ rt = 4.28 min

Example 6

(43) ##STR00046##

(44) Referring to Reaction Scheme 6, Stage 1, to a stirred solution of 6-(3-chloro-phenyl)-pyrimidine-4-carboxylic acid (1 eq) or 6-(3,4-dichloro-phenyl)-pyrimidine-4-carboxylic acid methyl ester in THF (20 vol) was added dropwise a 1M NaOH solution. The mixture was stirred at ambient temperature and the resulting precipitate was filtered and washed with water/THF or with water then heptane to furnish the described salts.

(45) TABLE-US-00006 Molecular Structure Weight Mass Spec Result 312.68 [M + H].sup.+ = 291/293, 100% @ rt = 3.97 min 383.81 [M + H].sup.+ = 362/364, 100% @ rt = 2.55 min 328.73 [M + H].sup.+ = 307/309, 100% @ rt = 4.35 min 0 307.67 [M + H]+ = 286/288, 99% @ rt = 3.26 min 385.8 [M ? Na + 2H]+ = 364/366 100% @ rt = 2.29 min 328.69 [M + H]+ = 307/309, 87% @ rt = 3.37 min

Example 7

(46) ##STR00053##

(47) Referring to Reaction Scheme 7, Stage 1, to a stirred suspension of 4-bromo-pyridine-2-carboxylic acid methyl ester (1 eq) in 1,4-dioxane (20 vol) was added the appropriate substituted phenyl boronic acid (1.1 eq) and Pd(PPh3)4 (0.05 eq). A 2M K2CO3 solution (7.5 vol) was added and the reaction mixture was heated at 90? C. with stirring for 16 hours under an atmosphere of N2. The reaction mixture was cooled to room temperature and the resulting precipitate was isolated by filtration to furnish the acid intermediate as the potassium salt, which was used without further purification in the stage. In the case of the 3-chlorophenyl analogue no precipitate was formed upon cooling, hence the solvent was removed in vacuo. The resulting residue was dissolved in EtOAc and water. Both phases were separated. EtOAc was removed in vacuo and the resulting residue was purified by flash column chromatography (eluent: [5:95] methanol:DCM) to furnish the desired 4-(3-chloro-phenyl)-pyridine-2-carboxylic acid methyl ester. The aqueous phase was acidified and the resulting precipitate was isolated by filtration and used as such in stage 2. Further purification was carried out by prep HPLC to furnish the required 4-(3-chloro-phenyl)-pyridine-2-carboxylic acid.

(48) Referring to Reaction Scheme 7, Stage 2, the required amide analogues were prepared following the procedure described in method A from 4-(3-chloro-phenyl)-pyridine-2-carboxylic acid, hydrochloride salt and were purified by trituration in acetonitrile/water (1/1) or in water followed by heptane.

(49) Referring to Reaction Scheme 7, Stage 3, the potassium salt isolated in stage 1 was suspended in HCl (2M) and stirred at ambient temperature for 2 hours. The solid was filtered and washed with water to furnish the desired target compound.

(50) Referring to Reaction Scheme 7, Stage 4, the required amide analogues were prepared following the procedure described in method A from 4-(substituted-phenyl)-pyridine-2-carboxylic acid potassium salt and were purified by trituration in acetonitrile/water (1/1) or in water followed by heptane.

(51) The following compounds were prepared substantially as described above.

(52) TABLE-US-00007 Structure Molecular Weight Mass Spec Result 289.72 [M + H].sup.+ = 290/292, 98% @ rt = 3.31 min 305.76 [M + H].sup.+ = 306/308, 99% @ rt = 3.73 min 305.76 [M + H].sup.+ 306/308, 99% @ rt = 3.71 min 291.74 [M + H].sup.+ = 292/294, 100% @ rt = 3.44 min

Example 8

(53) ##STR00058## ##STR00059##

(54) Referring to Reaction Scheme 8, Stage 1 a solution of NaNO2 (2.4 eq) in water (5 vol) was slowly added over 30 min to a suspension of [3-chloro-4-(trifluoromethoxy)phenyl]amine (1 eq) in (7 vol) of 15% HCl at ?5? C. The solid material was removed by filtration and a solution of NaBF4 (1.6 eq) in water (4 vol) was mixed with the filtrate. The resulting solid was collected by filtration, washed with minimum water and dried on a sinter funnel under vacuum for 1 hour. It was then dried in the vacuum oven at 40? C. until constant weight to give the required product.

(55) Referring to Reaction Scheme 8, Stage 2, 3-chloro-4-(trifluoromethoxy)benzene-1-diazonium tetrafluoroboranide (1 eq) was mixed with bis(pinacolato) diboron (1.05 eq) in a flask cooled by an ice bath. MeOH (8 vol) was added and the mixture was de-gassed with nitrogen for 10 minutes before PdCl2(dppf)2.DCM (0.025 eq) was added. The mixture was stirred at room temperature overnight before analysis by LCMS. The reaction was evaporated to dryness, re-dissolved in DCM, dry loaded onto silica and purified by dry flash chromatography running a slow gradient from 0-20% EtOAc in heptane. Clean fractions were combined and evaporated to dryness to give the required product as an oil.

(56) Referring to Reaction Scheme 8, Stage 3, 4,6-dichloropyrimidine (1 eq) and 2-[3-chloro-4-(trifluoromethoxy)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.7 eq) were dissolved in dioxane (12 vol) at room temperature and 2M potassium carbonate (2 eq) was added. The solution was degassed with nitrogen for 5 minutes. Pd(PPh3)4 (0.05 eq) was added and the reaction was stirred at 90? C. for 2 hours before analysis by LCMS. The reaction was cooled to room temperature and the solvent was evaporated. DCM was added and the organic layer was washed with water, brine and dried using MgSO4. The solvent was evaporated to dryness to give an oil which was purified by dry-flash chromatography eluting with 0-6% EtOAc in heptane. The resulting oil was dried in the vacuum oven at 40? C. to give the required product.

(57) Referring to Reaction Scheme 8, Stage 4, 4-Chloro-6-(3-chloro-4-trifluoromethoxy-phenyl)-pyrimidine (l eq), and triethylamine (2 eq) were dissolved in MeOH and degassed for 5 minutes with nitrogen. Pd(dppf)2Cl2.DCM (0.05 eq) was added and the reaction was sealed inside a 500 ml bomb. The bomb was charged with CO (5 bar) and heated at 50? C. overnight before analysis by LCMS. The reaction was cooled to room temperature and the solvent evaporated. The residue was re-dissolved in EtOAc and washed with water, brine and dried using MgSO4. The solvent was evaporated and the resulting solid purified by dry flash chromatography eluting with 30-40% EtOAc in heptane to give the required product.

(58) Referring to Reaction Scheme 8, Stage 5a, 6-(3-Chloro-4-trifluoromethoxy-phenyl)-pyrimidine-4-carboxylic acid methyl ester was dissolved in THF (16 vol) and 2M NaOH (2 eq) was added. The reaction mixture was allowed to stir at room temperature for 17 hours. Water (32 vol) was added and the mixture extracted with EtOAc (2?32 vol). 2 M HCl (2 eq) was added and the solution extracted with EtOAc (3?32 vol). The combined organic layers were dried over MgSO4 and the solvent removed to dryness. The crude compound was re-crystallised from acetonitrile (20 vol), filtered and dried in a vacuum oven at 40? C. to give the desired target 6-(3-chloro-4-trifluoromethoxy-phenyl)-pyrimidine-4-carboxylic acid.

(59) Referring to Reaction Scheme 8, Stage 5b, 6-(3-Chloro-4-trifluoromethoxy-phenyl)-pyrimidine-4-carboxylic acid methyl ester was dissolved in THF. 2M NaOH (2 eq) was added and the reaction was stirred at room temperature for 12 hours before analysis by LCMS. The reaction was evaporated to dryness and the resulting solid was washed with water and diethyl ether. The solid was dried in a vacuum oven at 40? C. to give the target compound 6-(3-chloro-4-trifluoromethoxy-phenyl)-pyrimidine-4-carboxylic acid as a sodium salt.

(60) The following compounds were prepared substantially as described above.

(61) TABLE-US-00008 Molecular Structure Weight Mass Spec Result 0 318.64 [M + H].sup.+ = 319/321, 74% @ rt = 4.32 min 340.62 [M + H].sup.+ = 319/321, 100% @ rt = 4.19 min

Example 9

(62) ##STR00062##

(63) Referring to Reaction Scheme 9, Stage 1 a solution of NaNO2 (2.4 eq) in water (5 vol) was slowly added over 30 min to a suspension of [3-chloro-4-(trifluoromethoxy)phenyl]amine (1 eq) in (7 vol) of 15% HCl at ?5? C. The solid material was removed by filtration and a solution of NaBF4 (1.6 eq) in water (4 vol) was mixed with the filtrate. The resulting solid was collected by filtration, washed with minimum water and dried on a sinter funnel under vacuum for 1 hour. It was then dried in the vacuum oven at 40? C. until constant weight to give the required product.

(64) Referring to Reaction Scheme 9, Stage 2, 3-chloro-4-(trifluoromethoxy)benzene-1-diazonium tetrafluoroboranide (1 eq) was mixed with bis(pinacolato) diboron (1.05 eq) in a flask cooled by an ice bath. MeOH (8 vol) was added and the mixture was de-gassed with nitrogen for 10 minutes before PdCl2(dppf)2.DCM (0.025 eq) was added. The mixture was stirred at room temperature overnight before analysis by LCMS. The reaction was evaporated to dryness, re-dissolved in DCM, dry loaded onto silica and purified by dry flash chromatography running a slow gradient from 0-20% EtOAc in heptane. Clean fractions were combined and evaporated to dryness to give the required product as an oil.

(65) Referring to Reaction Scheme 9, Stage 3, to a stirred suspension of 4-bromo-pyridine-2-carboxylic acid methyl ester (1 eq) in 1,4-dioxane (20 vol) was added the appropriate substituted phenyl boronic acid (1.1 eq) and Pd(PPh3)4 (0.05 eq). A 2M K2CO3 solution (7.5 vol) was added and the reaction mixture was heated at 90? C. with stirring for 16 hours under an atmosphere of N2. The reaction mixture was cooled to room temperature and the resulting precipitate was isolated by filtration to furnish the acid product as the potassium salt which was suspended in HCl (2M) and stirred at ambient temperature for 2 hours. The solid was filtered and washed with water to furnish the desired target compound.

(66) The following compounds were prepared substantially as described above.

(67) TABLE-US-00009 Molecular Structure Weight Mass Spec Result 317.65 [M + H].sup.+ = 317, 100% @ rt = 3.76 min

Example 10

(68) ##STR00064##

(69) Referring to Reaction Scheme 10, Stage 1. Sodium hydride (1.1 eq) was added portion wise to a cool (0? C.), stirred solution of 4-bromo-2-chlorophenol (1.0 eq) in DMF (6 vol) and the mixture stirred at this temperature under a nitrogen atmosphere for 30 minutes. After this time, 3-bromoprop-1-ene (1.1 eq) was added dropwise and the reaction mixture was allowed to warm to room temperature before being stirred at this temperature overnight. After this time, the reaction mixture was poured onto ice-water (10 vol), the mixture was extracted with ethyl acetate (3?), the organic layers were combined, washed with brine (5 vol), dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 20% ethyl acetate, 80% heptane) to give the desired compound as a yellow gum.

(70) Referring to Reaction Scheme 10, Stage 2. 1-Allyloxy-4-bromo-2-chloro benzene (l eq) was suspended in mesitylene (12 vol) and the mixture heated to 160? C. and stirred at this temperature overnight. After this time, the reaction mixture was cooled to room temperature and concentrated. The resulting residue was purified using a Biotage Isolera (340 g silica column eluting with a gradient from heptane to 100% DCM) to give the desired compound as a yellow oil.

(71) Referring to Reaction Scheme 10, Stage 3. Borane (1M solution in THF, 1 eq) was added drop wise to a stirred solution of 2-allyl-4-bromo-6-chloro-phenol (1 eq) in THF (10 vol) and the reaction mixture was stirred at room temperature under a nitrogen atmosphere for 4 hours. After this time, the reaction mixture was quenched by the sequential addition of water (1 eq), NaOH (1 eq) and hydrogen peroxide (1 eq) and the mixture stirred at room temperature for a further 2 hours. The resulting mixture was partitioned between diethyl ether (5 vol) and water (5 vol). The organic layer was separated, washed with brine (2 vol), dried (MgSO4), filtered and to give the desired compound as a colourless gum.

(72) Referring to Reaction Scheme 10, Stage 3. Diethyl diazene-1,2-dicarboxylate (1 eq) was added dropwise to a stirred solution of triphenyl phosphane (1 eq) and 4-bromo-2-chloro-6-(3-hydroxy-propyl)-phenol (1 eq) and the reaction mixture was stirred at room temperature under a nitrogen atmosphere overnight After this time, the reaction mixture was concentrated and purified using a Biotage Isolera (50 g silica column eluting with a gradient from 0% heptane to 20% ethyl acetate/80% heptane) to give the desired compound as a pale yellow oil.

(73) Referring to Reaction Scheme 10, Stage 4. Bis-pinacol borane (1.5 eq) was added in one portion to a cool (0? C.), stirred solution of 6-bromo-8-chloro-chroman (1.0 eq) and potassium acetate (3.5 eq) in DMSO (5 vol). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.1 eq) was added in one portion, the mixture was allowed to warm to room temperature and was stirred at this temperature under a nitrogen atmosphere for 1 hour. After this time the inorganic precipitate was removed by filtration and the filtrate was concentrated. The resulting residue was purified using a Biotage Isolera (50 g silica column eluting with a gradient from 0% heptane to 40% DCM/60% heptane) to give the desired compound as a pale yellow oil.

(74) Referring to Reaction Scheme 10, Stage 5. Tripotassium phosphate (2 eq) was added in one portion to a stirred solution of 8-chloro-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-chroman (1 eq) and methyl 4-bromopyridine-2-carboxylate (2 eq) in DMF (10 vol). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.2 eq) was added in one portion, the mixture was then heated to 60? C. and stirred at this temperature for 16 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and partitioned between ethyl acetate (5 vol) and water (5 vol). The organic layer was separated, washed sequentially with water (5 vol) then brine (5 vol) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (100 g silica column eluting with a gradient from 0% heptane to 80% DCM/20% heptane) to give the desired compound as a white solid.

(75) Referring to Reaction Scheme 10, Stage 5. 2M NaOH (4 eq) was added in one portion to a stirred solution of 6-(8-chloro-chroman-6-yl)-pyrimidine-4-carboxylic acid methyl ester (l eq) in ethanol (1 vol) and the mixture was stirred at room temperature for 2 hours. After this time the reaction mixture was diluted with water and the ethanol removed under reduced pressure. The remaining solution was acidified to pH 1 with 1M HCl and the resulting precipitate was collected by filtration, washed with water (5 vol) and TBME (5 vol) and dried in a vacuum oven at 40? C. overnight to afford the desired compound as a white solid.

(76) The following compounds were prepared substantially as described above.

(77) TABLE-US-00010 Molecular Structure Weight Mass Spec Result 290.71 [M + H]+ = 291, 100% @ rt = 3.71 min

Example 11

(78) ##STR00066##

(79) Referring to Reaction Scheme 11, Stage 1. Potassium carbonate (2 eq) was added portion wise to a stirred solution of 4-bromo-2-chlorophenol (1 eq) and bromoacetaldehyde diethyl acetal (1.5 eq) in DMF (6 vol) and the mixture was heated to 140? C. and heated at this temperature under a nitrogen atmosphere for 3 hours. After this time the reaction mixture was cooled to room temperature and concentrated. The resulting residue was partitioned between ethyl acetate (20 vol) and water (5 vol), the organic layer was separated, dried (MgSO4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (340 g silica column eluting with a gradient from 0% DCM to 60% DCM/40% heptane) to afford the desired compound as a colourless oil.

(80) Referring to Reaction Scheme 11, Stage 2. 4-Bromo-2-chloro-1-(2,2-diethoxy-ethoxy)-benzene (1 eq) was added portion wise as a solution in toluene (5 vol) to polyphosphonic acid (8 eq)) at 0? C. The resulting suspension was allowed to warm to room temperature before being heated to reflux and stirred for 1 hour. After this time the mixture was cooled to room temperature and partitioned between water (10 vol) and ethyl acetate (30 vol). The resulting residue was partitioned between ethyl acetate (30 vol) and water (5 vol), the organic layer was separated, dried (MgSO4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (340 g silica column eluting with 100% heptane) to afford the desired compound as a white solid.

(81) Referring to Reaction Scheme 11, Stage 3. Potassium acetate (3 eq) was added in one portion to a stirred solution of 5-bromo-7-chloro-benzofuran (1 eq) and bis-pinacol borane (1.1 eq) in DMF (3 vol). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.3 eq) was added in one portion, the mixture was then heated to 80? C. and stirred at this temperature for 18 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and partitioned between ethyl acetate (20 vol) and water (10 vol). The biphasic suspension was filtered through glass fiber filter paper and the organic layer was separated, washed sequentially with water (3?) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (100 g silica column eluting with 100% heptane to 50% DCM/50% heptane) to afford the desired compound as a white solid.

(82) Referring to Reaction Scheme 11, Stage 4. Tripotassium phosphate (1.4 eq) was added in one portion to a stirred solution of, 7-chloro-5-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzofuran (1 eq) and methyl 6-chloropyrimidine-4-carboxylate (2 eq) in DMF (4 vol). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.2 eq) was added in one portion, the mixture was then heated to 60? C. and stirred at this temperature for 16 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and partitioned between ethyl acetate (20 vol) and water (10 vol). The organic layer was separated, washed sequentially with water (10 vol) then brine (10 vol) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (50 g silica column eluting with 100% heptane to 20% ethyl acetate/50% heptane) to afford the desired compound as a white solid.

(83) Referring to Reaction Scheme 11, Stage 5. NaOH (1.5 eq) was added in one portion to a stirred solution of 6-(7-chloro-benzofuran-5-yl)-pyrimidine-4-carboxylic acid methyl ester (1.0 eq) in THF (8 vol) and the mixture was stirred at room temperature for 16 hours. After this time, the resulting precipitate was collected by filtration, washed with water (1 vol) and DCM (2 vol) before being dried under vacuum. This solid was then suspended in HCl (2M solution, 6 vol) and acetonitrile (6 vol), heated to 80? C. until complete dissolution then cooled to room temperature. The acetonitrile was removed under reduced pressure and the solid precipitate was collected by filtration, washed with water (1 vol) before being dried in a vacuum over overnight to give the hydrochloride salt of the desired compound as a white solid.

(84) The following compounds were prepared substantially as described above.

(85) TABLE-US-00011 Molecular Mass Spec Structure Weight Result 274.67 [M + H]+ = 275/277, 98% @ rt = 3.70 min

Example 12

(86) ##STR00068##

(87) Referring to Reaction Scheme 12, Stage 1. Potassium carbonate (2M solution, 52.0 ml, 104.0 mmol) was added in one portion to a stirred solution of 3,4-dichlorophenyl boronic acid (6.9 g, 37.0 mmol) and 4,6-dichloro-5-methyl pyrimidine (8.5 g, 52.0 mmol) in dioxane (150 ml). The mixture was degassed with nitrogen for 5 minutes, after which time palladium tetrakis triphenylphosphine (3.0 g, 3.0 mmol) was added in one portion, the mixture was then heated to 90? C. and stirred at this temperature for 16 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and concentrated. The resulting residue was dissolved in DCM (500 ml), washed sequentially with water (500 ml) then brine (500 ml) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 6% EtOAc, 94% Heptane) to give the desired compound (6.05 g, 42% yield) as a white solid. 6H (500 MHz, DMSO) 8.91-9.00 (1H, m) 7.88-7.96 (1H, m) 7.76-7.88 (1H, m) 7.58-7.69 (1H, m) 2.36 (3H, s). Tr=2.30 min m/z (ES+) (M+H+) 275, 277.

(88) Referring to Reaction Scheme 12, Stage 2. Triethylamine (6.1 ml, 44.0 mmol) was added in one portion to a calorimeter containing a stirred solution of 4-chloro-6-(3,4-dichloro-phenyl)-5-methyl-pyrimidine (5.95 g, 22.0 mmol) in methanol (80 ml). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.9 g, 1.0 mmol) was added in one portion, the calorimeter was sealed, pressurised with carbon monoxide (5 bar) and was heated to 50? C. overnight. After this time the reaction mixture was cooled to room temperature, diluted with methanol and concentrated. The resulting residue was dissolved in DCM (300 ml) and washed sequentially with water (250 ml) and brine (250 ml). The organic layer was separated, dried (MgSO4), filtered, concentrated and the resulting residue purified by flash column chromatography (elution: 40% EtOAc, 60% heptane) to give the desired compound (5.2 g, 80% yield) as a white solid. ?H (500 MHz, DMSO) 9.19 (1H, s) 7.92-7.97 (1H, m) 7.79-7.85 (1H, m) 7.63-7.70 (1H, m) 3.95 (3H, s) 2.30-2.42 (3H, m). Tr=2.10 min m/z (ES+) (M+H+) 297, 299.

(89) Referring to Reaction Scheme 12, Stage 3. NaOH (2M solution, 1.1 ml, 2.0 mmol) was added in one portion to a stirred solution of 6-(3,4-dichloro-phenyl)-5-methyl-pyrimidine-4-carboxylic acid methyl ester (0.32 g, 1.0 mmol) in THF (10 ml) and the mixture was stirred at room temperature for 16 hours. After this time, the resulting precipitate was collected by filtration, washed with water (1 ml) and DCM (20 ml) before being dried under vacuum. This solid was then suspended in HCl (2M solution, 60 ml) and acetonitrile (60 ml), heated to 80? C. until complete dissolution then cooled to room temperature. The acetonitrile was removed under reduced pressure and the solid precipitate was collected by filtration, washed with water (10 ml) before being dried in a vacuum over overnight to give the hydrochloride salt of the desired compound (0.22 g, 75% yield) as a white solid.

(90) The following compounds were prepared substantially as described above.

(91) TABLE-US-00012 Molecular Structure Weight Mass Spec Result 304.72 [M + H]+ = 305/307, 100% @ rt = 3.64 min

Example 13

(92) ##STR00070##

(93) Referring to Reaction Scheme 13, Stage 1. Sodium bicarbonate (0.46 g, 5.0 mmol) was added in one portion to a stirred solution of 5-bromomethyl-6-(3,4-dichloro-phenyl)-pyrimidine-4-carboxylic acid methyl ester (0.24 g, 0.64 mmol) in DMSO (5 ml), and the mixture was stirred at room temperature under a nitrogen atmosphere for 20 hours. After this time the mixture was partitioned between ethyl acetate (20 ml) and water (20 ml), the organic layer was separated and the aqueous layer extracted with ethyl acetate (2?20 ml). The organic layers were combined, dried (MgSO4), filtered, concentrated and the resulting residue was triturated with diethyl ether. The resulting precipitate was collected by filtration and dried under vacuum to give the desired compound (0.08 g, 45% yield) as an orange solid.

(94) Referring to Reaction Scheme 13, Stage 2. Sodium methoxide (0.02 g, 0.36 mmol) was added in one portion to a stirred solution of 4-(3,4-dichloro-phenyl)-5H-furo[3,4-d]pyrimidin-7-one (0.05 g, 0.18 mmol) in methanol (5 ml), and the mixture was stirred at room temperature under a nitrogen atmosphere for 20 hours. After this time, sodium hydroxide (2M solution, 0.05 ml, 0.89 mmol) was added and the mixture was heated to 70? C. and stirred at this temperature for a further 4 hours. After this time the reaction mixture was cooled to room temperature and the resulting precipitate was collected by filtration, washed with methanol (5 ml) and dried under vacuum to give the desired compound (0.01 g, 5% yield) as an off-white solid.

(95) Referring to Reaction Scheme 13, Stage 3. Sodium methoxide (0.03 g, 0.53 mmol) was added in one portion to a stirred solution of 5-bromomethyl-6-(3,4-dichloro-phenyl)-pyrimidine-4-carboxylic acid methyl ester (0.1 g, 0.26 mmol) in methanol (5 ml), and the mixture was stirred at room temperature under a nitrogen atmosphere for 20 hours. After this time the mixture was concentrated and the resulting residue taken up in DCM (10 ml). The solution was washed consecutively with water (2?50 ml) and brine (2?50 ml), before being separated, dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 100% DCM to 99% DCM: 1% Methanol) to give the desired compound (0.02 g, 20% yield) as a white solid. Tr=2.11 min m/z (ES+) (M+H+) 327, 329.

(96) Referring to Reaction Scheme 13, Stage 4. Sodium hydroxide (0.05 ml, 0.1 mmol) was added in one portion to a stirred solution of methyl 6-(3,4-dichlorophenyl)-5-(methoxymethyl)pyrimidine-4-carboxylate (0.1 g, 0.26 mmol) in THF (5 ml) and the mixture was stirred at room temperature under a nitrogen atmosphere for 20 hours. After this time the resulting precipitate was collected by filtration, washed with water (1 ml) and dried under vacuum to give the desired compound (0.004 g, 15% yield) as a white solid.

(97) The following compounds were prepared substantially as described above.

(98) TABLE-US-00013 Molecular Structure Weight Mass Spec Result 302.72 [M + H]+ = 303/305, 100% @ rt = 4.20 min 320.72 [M + H]+ = 321/323, 100% @ rt = 3.29 min 316.75 [M + H]+ = 317/319, 100% @ rt = 3.89 min

Example 14

(99) ##STR00074##

(100) Referring to Reaction Scheme 14, Stage 1. 2,2-Dimethylpropanoyl chloride (0.07 ml, 0.53 mmol) was added dropwise to a stirred solution of 6-(3-chloro-4-cyclopropoxy-phenyl)-pyrimidine-4-carboxylic acid (0.15 g, 0.48 mmol) in THF (10 ml) and the mixture was stirred at room temperature for 2 hours. After this time the mixture was added portion wise to a solution of (1R)-1-[(3aR,5R,6S,6aR)-6-hydroxy-2,2-dimethyl-tetrahydro-2H-furo[2,3-d][1,3]dioxol-5-yl]ethane-1,2-diol (0.32 g, 1.44 mmol) in pyridine (10 ml) and the reaction mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. The resulting mixture was concentrated and the residue partitioned between DCM (50 ml) and water (20 ml). The organic layer was separated, dried (MgSO4), filtered and concentrated. The resulting residue was then purified by flash column chromatography (elution: 100% ethyl acetate) to give the desired compound (0.095 g, 34% yield) as a colourless oil. Tr=1.95 min m/z (ES+) (M+H+) 493.

(101) Referring to Reaction Scheme 14, Stage 2. 4M HCl in dioxane solution (5 ml) was added in one portion to a stirred solution of 6-(3-chloro-4-cyclopropoxy-phenyl)-pyrimidine-4-carboxylic acid 6-hydroxy-2,2-dimethyl-tetrahydro-furo[2,3-d][1,3]dioxol-5-ylmethyl ester (0.095 g, 0.19 mmol) in dioxane (2 ml) and the mixture was stirred at room temperature overnight. The resulting mixture was concentrated and the resulting residue was then purified by prep HPLC to give the title compound (0.01 g, 13% yield) as a colourless glass.

(102) TABLE-US-00014 Molecular Structure Weight Mass Spec Result 452.85 [M + Na]+ = 475.0 @ rt = 3.36 + 3.41 min

Example 15

(103) ##STR00076##

(104) Referring to Reaction Scheme 15, Stage 1. Triethylamine (19.01 ml, 146.92 mmol) was added dropwise to a solution of diethyl but-2-ynedioate (25.0 g, 146.92 mmol) and formamidine hydrochloride (11.83 g, 146.92 mmol) in acetonitrile (500 mL). The resulting red solution was heated at 80? C. for 2.5 hours. After this time the reaction mixture was cooled to 5? C. using a saturated NaCl/ice bath and the reaction was stirred at this temperature for 25 minutes. After this time the resulting solid precipitate was collected under suction and dried on a sinter funnel for 30 minutes under vacuum at room temperature before drying in the vacuum oven at room temperature for 3 hours to give the desired compound (21.3 g, 86% yield) as a pale brown solid. Tr=0.85 min (3.5 minute method) m/z (ES+) (M+H+) 169.

(105) Referring to Reaction Scheme 15, Stage 2. Ethyl 6-hydroxypyrimidine-4-carboxylate (21.3 g, 126.67 mmol) was dissolved in dry DMF (100 mL) in a 2 neck flask. The flask was purged with a stream of nitrogen while cooling in an ice bath for 10 minutes. After this time, thionyl chloride (15.6 mL, 215.6 mmol) was added dropwise over 20 minutes, before being warmed to room temperature and stirred under a nitrogen atmosphere for 2 hours. After this time, the reaction mixture was carefully poured onto ?100 mL ice water. TBME (100 mL) was added, the organic layer was separated and the aqueous extracted with further TBME (3?100 mL). The combined organic layers were washed consecutively with water (2?100 mL), and brine (100 mL) before being dried (MgSO4), filtered and concentrated to give the desired compound (8.8 g, 37% yield) as a light orange powder. ?H (500 MHz, DMSO) 9.23 (d, J=0.95 Hz, 1H), 8.16 (d, J=1.10 Hz, 1H), 4.39 (q, J=7.09 Hz, 2H), 1.34 (t, J=7.17 Hz, 3H). Tr=1.43 min (3.5 minute method) m/z (ES+) (M+H+) 187.

(106) Referring to Reaction Scheme 15, Stage 3. Tripotassium phosphate (1.12 g, 5.63 mmol) was added in one portion to a stirred solution of 2-(2H-1,3-benzodioxol-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.93 g, 3.75 mmol) and ethyl 6-chloropyridine-4-carboxylate (0.7 g, 3.75 mmol) in DMF (20 mL). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.14 g, 0.19 mmol) was added in one portion, the mixture was then heated to 80? C. and stirred at this temperature for 16 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and partitioned between ethyl acetate (200 mL) and water (100 mL). The organic layer was separated, washed sequentially with water (100 mL) then brine (100 mL) before being dried (MgSO4), filtered and concentrated. The resulting brown solid was purified by flash column chromatography (elution: 40% EtOAc, 60% Heptane) to give the desired compound (0.31 g, 31% yield) as a white solid. Tr=1.87 min m/z (ES+) (M+H+) 273.

(107) Referring to Reaction Scheme 15, Stage 4. NaOH (2M solution, 0.63 mL, 1.27 mmol) was added in one portion to a stirred solution of ethyl 6-(2H-1,3-benzodioxol-5-yl)pyrimidine-4-carboxylate (0.31 g, 1.15 mmol) in THF (10 mL) and the mixture was stirred at room temperature for 16 hours before being heated to reflux for 2 hours. After this time, the reaction mixture was cooled to room temperature and the resulting precipitate was collected by filtration, washed with THF (20 mL) before being dried under vacuum to give the desired compound (0.17 g, 56% yield, >99% purity) as a white solid.

(108) The following compounds were prepared substantially as described above.

(109) TABLE-US-00015 Molecular Structure Weight Mass Spec Result 244.04 [M + H]+ = 245/247, 99% @ rt = 3.08 min 280.73 [M + H]+ = 281/283, 99% @ rt = 2.61 min 278.04 [M + H]+ = 279/281, 100% @ rt = 3.65 min 0 286.2 [M + H]+ = 287/289, 100% @ rt = 3.03 min

Example 16

(110) ##STR00081##

(111) Referring to Reaction Scheme 16, Stage 1. A solution of oxone (0.25 g, 0.40 mmol) in water (12 mL) was added portion wise over 15 minutes to a stirred solution of ethyl 6-[3-chloro-4-(methylsulfanyl)phenyl]pyrimidine-4-carboxylate (0.25 g, 81 mmol) in acetone (12 mL) and the resulting mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. After this time, the reaction was partitioned between water (20 mL) and ethyl acetate (50 mL). The organic layer was separated, and the aqueous further extracted with ethyl acetate (2?50 mL). The combined organic extracts were then dried (MgSO4), filtered and concentrated. The resulting residue was purified on a Biotage isolera (15% ethyl acetate, 90% heptanes to 100% ethyl acetate) to give the desired compound (0.2 g, 76% yield) as a white solid. ?H (500 MHz, DMSO-d6) 9.48 (d, J=1.20 Hz, 1H), 8.66 (d, J=1.22 Hz, 1H), 8.56 (dd, J=1.64, 8.22 Hz, 1H), 8.48 (d, J=1.58 Hz, 1H), 8.02 (d, J=8.21 Hz, 1H), 4.43 (q, J=7.11 Hz, 2H), 2.87 (s, 3H), 1.38 (t, J=7.11 Hz, 3H). Tr=1.64 min m/z (ES+) (M+H+) 325, 327.

(112) Referring to Reaction Scheme 16, Stage 2. NaOH (2M solution, 0.33 mL, 0.66 mmol) was added in one portion to a stirred solution of ethyl 6-(3-chloro-4-methanesulfinylphenyl)pyrimidine-4-carboxylate (0.19 g, 0.61 mmol) in THF (30 mL) and the mixture was stirred at room temperature for 7 hours. After this time, the resulting precipitate was collected by filtration, washed with THF (10 mL) before being dried under vacuum to give the desired compound (0.17 g, 84% yield, >99% purity) as a white solid.

(113) The following compounds were prepared substantially as described above.

(114) TABLE-US-00016 Molecular Mass Spec Structure Weight Result 296.73 [M + H]+ = 297/299 98.9% @ rt = 2.83 min 312.73 [M + H]+ = 313/315 100% @ rt = 2.92 min

Example 17

(115) ##STR00084##

(116) Referring to Reaction Scheme 17, Stage 1. Cyclopropylmagnesium bromide (0.5M solution in THF, 100.0 mL, 50.0 mmol) was added portion wise over 1 hour to a cold (?78? C.), stirred solution of 4-bromo-2-chlorobenzaldehyde (5.5 g, 25.0 mmol) in THF (100 mL) and the mixture was stirred for 1 hour before being allowed to warm to room temperature and stirred for a further 18 hours. After this time, the reaction was quenched by the addition of saturated ammonium chloride (100 mL) and the mixture extracted with ethyl acetate (3?100 mL). The combined organic extracts were combined, washed with water (100 mL) and brine (100 mL) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 10% ethyl acetate, 90% heptanes) to give the desired compound (5.05 g, 77% yield) as a pale yellow oil. 6H (500 MHz, DMSO) 7.66 (d, J=1.89 Hz, 1H) 7.50-7.60 (m, 2H) 5.43 (br. s., 1H) 4.59 (d, J=5.20 Hz, 1H) 1.04-1.15 (m, 1H) 0.29-0.46 (m, 4H).

(117) Referring to Reaction Scheme 17, Stage 2. Potassium acetate (3.72 g, 40.0 mmol) was added in one portion to a stirred solution of (4-bromo-2-chlorophenyl)(cyclopropyl)methanol (3.3 g, 1.3 mmol) and bis-pinacol borane (3.85 g, 1.5 mmol) in DMSO (35 mL). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.46 g, 0.6 mmol) was added in one portion, the mixture was then heated to 80? C. and stirred at this temperature for 16 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and partitioned between ethyl acetate (100 mL) and water (50 mL). The biphasic suspension was filtered through glass fiber filter paper and the organic layer was separated, washed sequentially with water (3?100 mL) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 80% heptane, 20% DCM and 2 mL of triethylamine) to give the desired compound (3.5 g, 90% yield) as a colourless oil. 6H (500 MHz, DMSO) 7.61 (s, 2H) 7.56 (s, 1H) 5.39 (d, J=4.41 Hz, 1H) 4.66 (t, J=5.20 Hz, 1H) 1.24-1.36 (m, 12H) 1.05-1.12 (m, 1H) 0.24-0.47 (m, 4H).

(118) Referring to Reaction Scheme 17, Stage 3. Tripotassium phosphate (1.03 g, 4.8 mmol) was added in one portion to a stirred solution of [2-chloro-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl](cyclopropyl)methanol (1.0 g, 3.2 mmol) and ethyl 6-chloropyrimidine-4-carboxylate (0.73 g, 3.89 mmol) in DMF (20 mL). The mixture was degassed with nitrogen for 5 minutes, after which time Pd(dppf)2Cl2 (0.13 g, 0.16 mmol) was added in one portion, the mixture was then heated to 60? C. and stirred at this temperature for 16 hours under a nitrogen atmosphere. After this time the reaction mixture was cooled to room temperature and partitioned between ethyl acetate (100 mL) and water (50 mL). The organic layer was separated, washed sequentially with water (50 mL) then brine (50 mL) before being dried (MgSO4), filtered and concentrated. The resulting red gum was purified by flash column chromatography (elution: 40% EtOAc, 60% Heptane) to give the desired compound (0.74 g, 65% yield) as a colourless oil. ?H (500 MHz, DMSO) 9.42 (d, J=1.10 Hz, 1H) 8.57 (d, J=1.10 Hz, 1H) 8.22-8.36 (m, 2H) 7.79 (d, J=8.20 Hz, 1H) 5.52 (br. s., 1H) 4.72 (d, J=5.99 Hz, 1H) 4.43 (q, J=7.09 Hz, 2H) 1.38 (t, J=7.09 Hz, 3H) 1.15-1.22 (m, 1H) 0.29-0.53 (m, 4H). Tr=2.27 min m/z (ES+) (M+H+) 321.

(119) Referring to Reaction Scheme 17, Stage 4. NaOH (2M solution, 0.24 mL, 0.48 mmol) was added in one portion to a stirred solution of ethyl 6-{3-chloro-4-[cyclopropyl(hydroxy)methyl]phenyl}pyrimidine-4-carboxylate (0.16 g, 0.48 mmol) in THF (2 mL) and the mixture was stirred at room temperature for 16 hours. After this time, the resulting precipitate was collected by filtration, washed with water (1 mL) and DCM (20 mL) before being dried under vacuum to give the desired compound (0.065 g, 41% yield) as a white solid.

(120) Referring to Reaction Scheme 17, Stage 5. Dess-Martin Periodinane (0.36 g, 1.08 mmol) was added portion wise to a cooled (0? C.), stirred solution of 6-{3-chloro-4-[cyclopropyl(hydroxy)methyl]phenyl}pyrimidine-4-carboxylic acid (0.36 g, 1.08 mmol) in DCM (3 mL) and the mixture was allowed to warm to room temperature and stirred for 18 hours. After this time, the mixture was partitioned between DCM (20 mL) and saturated sodium bicarbonate (20 mL). The organic layer was separated, washed with water (100 mL) and brine (50 mL) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 20% ethyl acetate, 80% heptanes) to give the desired compound (0.26 g, 74% yield) as a white solid.

(121) The following compounds were prepared substantially as described above.

(122) TABLE-US-00017 Structure Molecular Weight Mass Spec Result 304.74 [M + H]+ = 305/307, 98% @ rt = 3.25 min 302.72 [M + H]+ = 303/305, 100% @ rt = 3.54 min

Example 18

(123) ##STR00087##

(124) Referring to Reaction Scheme 18, Stage 1. To a stirred solution of 4-bromo-2-chlorobenzaldehyde (0.51 g, 2.32 mmol) in a mixture of dry dioxane (2.5 mL) and dry DMF (0.60 mL) was added 4,4,4,4,5,5,5,5-octamethyl-2,2-bi-1,3,2-dioxaborolane (0.64 g, 2.52 mmol) and potassium acetate (0.7 g, 7.13 mmol). The mixture was degassed and then 1,1-bis(diphenylphosphanyl)ferrocene-dichloropalladium (1:1) (0.08 g, 0.11 mmol) was added. The mixture was further degassed before heating to 80? C. for 3 hours under an atmosphere of nitrogen gas. To the cooled reaction mixture was added water (30 mL) and EtOAc (15 mL); the organic layer was then washed with a 3:1 mixture of water and brine (2?40 mL), brine (5 mL), dried (MgSO4), filtered and concentrated. The resulting residue was then absorbed onto silica gel (1.6 g) and purified by dry flash chromatography (0-20% EtOAc in heptane) to give the desired compound (0.25 g, 37% yield @ 90% NMR purity) as a white partial solid. Tr=1.46 min (63%) & 2.45 min (30%) m/z (ES+) (M+H+) no ionisation.

(125) Referring to Reaction Scheme 18, Stage 2. To a degassed stirred solution of ethyl 6-chloropyrimidine-4-carboxylate (0.17 g, 0.9 mmol) and 2-chloro-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde (0.22 g, 0.81 mmol) in dioxane (2.5 mL) was added 2M K2CO3 (1.25 mL). Pd(PPh3)4 (57 mg, 0.05 mmol) was then added and the reaction mixture was further degassed before heating to 90? C. under an atmosphere of nitrogen gas for 2 hours. After this time, the reaction mixture was cooled to room temperature and concentrated. Water (5 mL) was then added and the solid filtered, washed with water (2 mL), acetone (3?2 mL) and dried under vacuum. The solid was suspended in a mixture of EtOAc (30 mL) and 1N HCl (10 mL) and then heated to achieve partial solution. The cooled two-phase system was then sonicated to achieve full dissolution. The aqueous layer was re-extracted with EtOAc (10 mL); the combined organics were washed with brine (5 mL), dried (MgSO4), filtered and concentrated to give the desired compound (0.1 g, 42% yield @ 85% purity) as a beige solid. Tr=1.58 min m/z (ES+) (M+H+) 263/265.

(126) Referring to Reaction Scheme 18, Stage 3. To a stirred suspension of 6-(3-chloro-4-formylphenyl)pyrimidine-4-carboxylic acid (93 mg, 0.35 mmol) in 1,2-dichloroethane (5 mL) was added dimethylamine (2M solution in THF, 0.53 mL) at room temperature followed by molecular sieves and sodium triacetoxyborohydride (125 mg, 0.59 mmol). After 1.5 hours, acetic acid (31 ?l, 0.54 mmol) was added and the reaction stirred at room temperature for 2.5 days. Further dimethylamine (2M solution in THF, 1.0 mL) and sodium triacetoxyborohydride (130 mg) were added and the mixture stirred for 6 h before a further amount of dimethylamine (2M in THF, 1.0 mL), sodium triacetoxyborohydride (130 mg) and AcOH (62 L). The mixture was then stirred for 18 hours. The reaction mixture was filtered and the filtrate was concentrated. A solution of 1:1 (v/v) MeCN:water (0.5 mL) was added to the resulting residue and then concentrated HCl (0.5 mL) was added dropwise. The crude product dissolved and was purified by preparative HPLC (acetonitrile and water) to give 14 mg of an off-white solid. The solid was further purified by sonication in TBME (1 mL) and collected by filtration. The solid was washed with TBME (4?1 mL) and dried to give the desired compound (7.8 mg, 7.9% yield @ 95% purity) as a off-white solid.

(127) The following compounds were prepared substantially as described above.

(128) TABLE-US-00018 Mole- cular Mass Spec Structure Weight Result 291.74 [M + H]+ = 292/294, 100% @ rt = 2.00 min

Example 19

(129) ##STR00089##

(130) Referring to Reaction Scheme 19, Stage 1. 4-Bromo-2-chloroaniline (2.0 g, 9.69 mmol), 1,4-dibromobutane (2.31 ml, 19.4 mmol), potassium carbonate (2.68 g, 19.4 mmol), water (25 mL) and dioxane (10 mL) were heated to 100? C. overnight with vigorous stirring. The reaction mixture was allowed to cool then extracted with EtOAc (2?25 mL). The combined organics were washed with brine (15 mL), dried (MgSO4), filtered and concentrated to give an orange oil. Column chromatography (Elution: 0-20% EtOAc-heptane) afforded the desired compound (1.16 g, 45% yield) as a yellow oil. ?H (500 MHz, DMSO-d6) 7.48 (d, J=2.36 Hz, 1H), 7.33 (dd, J=2.36, 8.83 Hz, 1H), 6.87 (d, J=8.83 Hz, 1H), 3.29-3.33 (m, 4H), 1.87 (td, J=3.43, 6.38 Hz, 4H); Tr (3 min)=2.68 min m/z (ES+) (M+H)+260, 262.

(131) Referring to Reaction Scheme 19, Stage 2. Potassium acetate (1.31 g, 13.4 mmol), bis(pinacolato)diboron (1.36 g, 5.32 mmol) and 1-(4-bromo-2-chlorophenyl)pyrrolidine (1.16 g, 4.45 mmol) were suspended in DMSO (15 mL). The solution was degassed with N2 for 5 min. PdCl2(dppf) (0.16 g, 0.22 mmol) was added and the reaction mixture was heated to 80? C. for 3 h. The reaction was cooled to rt. Water (30 mL) was added to the reaction and the aqueous was extracted using EtOAc (5?20 mL). The combined organic layers were washed with water (100 mL), brine (50 mL), dried (MgSO4), filtered, and concentrated to give a black oil. Column chromatography (Elution; 8% EtOAc-heptane) afforded the desired compound (1.14 g, 83% yield) as a pale yellow oil. Tr (3 min)=2.70 min m/z (ES+) (M+H)+307.

(132) Referring to Reaction Scheme 19, Stages 3 & 4 were carried out as described in Reaction Scheme 15.

(133) The following compounds were prepared substantially as described above.

(134) TABLE-US-00019 Molecular Mass Spec Structure Weight Result 0 303.74 [M + H]+ = 304/306, 100% @ rt = 4.14 min

Example 20

(135) ##STR00091##

(136) Referring to Reaction Scheme 20, Stage 1. In a three neck flask with dropping funnel, thermometer and nitrogen bubbler (no nitrogen input), 4-bromo-2-chlorophenol (5.0 g, 0.024 mol) was fully dissolved in acetic acid (25 mL) at room temperature. Nitric acid (70%, 2.9 mL, 0.048 mol) was added slowly dropwise over approx 15 minutes keeping the temperature at below 30? C. The reaction turned orange with an orange precipitate. The reaction was stirred for a further 4 hours at 20? C. After this time the reaction mixture was cautiously transferred via pipette onto approximately 50 mL ice. Once the ice had melted the yellow precipitate was filtered and washed with water (50 mL). The yellow solid was air dried under vacuum for 1 hour before being dissolved in DCM and dry loaded onto 5.5 g silica. The compound was purified by flash column chromatography (elution; 100% heptane, to 20% DCM in heptane, to 40% DCM in heptane, to 50% DCM in heptanes) to give the desired compound (4.38 g, 72% yield @ 100% UV purity) as a yellow solid. Tr=1.97 min m/z (ES+) no ionisation.

(137) Referring to Reaction Scheme 20, Stage 2. 4-Bromo-6-chloro-2-nitrophenol (4.38 g, 17.35 mmol) was dissolved in ethanol (120 mL). Water (28 mL) and saturated aqueous ammonium chloride (28 mL) were added followed by iron powder (7.75 g, 139 mmol). The reaction was heated to 50? C. and stirred for 1 hour, after which time the reaction was cooled to room temperature and filtered through a pad of celite (approx. 5 cm in a Jones tube), washing with 50 mL EtOH followed by excess EtOAc until the liquid ran clear. The organic layer was washed with water (50 mL). The water was re-extracted with EtOAc (2?200 mL). The combined organic extracts were washed with brine (20 mL), dried (MgSO4), filtered and concentrated. The resulting residue was dry loaded onto 5 g silica and purified by flash column chromatography (elution; 0-30% EtOAc in heptanes) to give the desired compound (2.76 g, 72% yield @ 100% UV purity) as a pale brown solid. Tr=1.65 min m/z (ES+) (M+H+) 222/224/226.

(138) Referring to Reaction Scheme 20, Stage 3. 2-Amino-4-bromo-6-chlorophenol (2.66 g, 11.96 mmol) was dissolved in triethylorthoacetate (24 mL). pTSA monohydrate (0.068 g, 0.359 mmol) was added and the reaction was stirred at 140? C. overnight. After this time the reaction was cooled to room temperature and the resulting solid was collected by filtration and dried under suction at room temperature for 2 hours to give the title compound (1.58 g, 54% yield @ 100% UV purity) as a white solid. Tr=2.07 min m/z (ES+) (M+H+) 246/248.

(139) Referring to Reaction Scheme 20, Stages 4, 5 & 6 were carried out as described in Reaction Scheme 15.

(140) The following compounds were prepared substantially as described above.

(141) TABLE-US-00020 Molecular Mass Structure Weight Spec Result 289.68 [M + H]+ = 290/292, 100% @ rt = 3.42 min

Example 21

(142) ##STR00093##

(143) Referring to Reaction Scheme 21, Stage 1. A solution of 4-bromo-2-chlorophenol (10.0 g, 48.0 mmol) in anhydrous DMF (30 mL) was added to a stirred suspension of sodium hydride (2.31 g, 58.0 mmol) in DMF (20 mL) cooled to 0? C. under nitrogen over 15 min, and stirring continued for 30 min. 3-Bromoprop-1-ene (7.00 g, 58.0 mmol) was added dropwise at 0? C. After 1 h, the mixture was allowed to warm to room temperature and then stirred for 3 d. Aqueous saturated NH4Cl (50 mL) was added over 10 min with ice-cooling, and the mixture was concentrated. The residue was treated with water (100 mL) and the mixture extracted with ethyl acetate (3?120 mL). The combined, dried (Na2SO4) organic extracts were concentrated to give an oil which contained DMF. A solution of the oil in ethyl acetate (100 mL) was washed with water (100 mL) and the dried (Na2SO4) organic layer was concentrated to give the desired compound (11.6 g, 87% yield) as a colourless oil. ?H (500 MHz, CDCl3) 7.50 (d, J=2.40 Hz, 1H), 7.30 (dd, J=2.40, 8.77 Hz, 1H), 6.79 (d, J=8.78 Hz, 1H), 6.04 (ddt, J=5.10, 10.38, 17.14 Hz, 1H), 5.45 (dd, J=1.44, 17.26 Hz, 1H), 5.32 (dd, J=1.33, 10.57 Hz, 1H), 4.59 (d, J=5.10 Hz, 2H).

(144) Referring to Reaction Scheme 21, Stage 2. A solution of 1-allyloxy-4-bromo-2-chloro-benzene (90%, 11.6 g, 42 mmol) in mesitylene (200 mL) was heated under nitrogen for 48 h at 190? C. with stirring. The reaction was concentrated and purified by column chromatography (Elution: 0-10% EtOAc-heptane) to afford the desired compound (4.66 g, 36% yield) as a colourless oil. Tr (3 min)=2.22 min m/z (ES+) (M+H+) 245, 247.

(145) Referring to Reaction Scheme 21, Stage 3. Sodium periodate (9.04 g, 42.3 mmol) was added to a stirred mixture of 2-allyl-4-bromo-6-chloro-phenol (5.23 g, 21.1 mmol), THF (100 mL) and water (100 mL) at room temperature. After 5 min, osmium tetroxide (13.5 ml of a 0.157 M solution in water, 2.1 mmol) was added and stirring continued for 1.5 h. The mixture was poured into brine (100 mL) and extracted with ethyl acetate (2?100 mL) and the combined, dried (Na2SO4) organic extracts were concentrated to give a dark oil. A stirred solution of the dark oil in methanol (100 mL) under nitrogen was cooled to 0? C., and treated with sodium borohydride (2.40 g, 63.4 mmol) in small portions over 20 min, maintaining the temperature between 0 and 10? C. After stirring for 16 h, the mixture was concentrated, treated with aqueous 1M hydrochloric acid (80 mL) and extracted with ethyl acetate (2?100 mL). The combined, dried (Na2SO4) organic extracts were concentrated, and the residue purified by column chromatography (Elution: 5-40% EtOAc-heptane) to afford the desired compound (1.60 g, 27% yield) as a colourless oil. Tr (3 min)=1.81 min m/z (ES+) (M+H+) 249, 251.

(146) Referring to Reaction Scheme 21, Stage 4. DIAD (1.52 ml, 7.70 mmol) was added to a stirred solution of 4-bromo-2-chloro-6-(2-hydroxy-ethyl)-phenol (1.49 g, 5.92 mmol) and triphenylphosphine (2.02 g, 7.70 mmol) in dry THF (1.5 mL) under nitrogen, with ice-cooling. After stirring for 16 h at rt, the solution was evaporated and the residual oil purified by column chromatography (Elution: 0-10% EtOAc-heptane) afforded the desired compound (1.20 g, 68% yield) as a colourless oil. Tr (3 min)=2.27 min m/z (ES+) no ionization.

(147) Referring to Reaction Scheme 21, Stages 5, 6 & 7 were carried out as described in Reaction Scheme 15.

(148) The following compounds were prepared substantially as described above.

(149) TABLE-US-00021 Molecular Mass Spec Structure Weight Result [M + H]+ = 277/279, 100% @ rt = 3.53 min

Example 22

(150) ##STR00095##

(151) Referring to Reaction Scheme 22, Stage 1. 4-Bromo-2-chlorophenol (14.0 g, 0.067 mol) was dissolved in acetic acid (75 mL) at room temperature. Nitric acid (70%, 8.00 ml, 0.145 mol) was added dropwise over approx 30 min keeping the temperature at roughly 20-22? C. After 1 h at rt, the reaction mixture was cautiously transferred via pipette onto approx 100 mL ice. Once the ice had melted the yellow precipitate was filtered, washing with a very small volume of water. The yellow solid was dried under suction. Purification by dry flash chromatography (Elution: 0-50% DCM-heptane) afforded the desired compound (12.0 g, 70% yield) as a yellow powder. ?H (500 MHz, DMSO) 11.35 (br. s., 1H) 8.09 (d, J=2.52 Hz, 1H) 8.07 (d, J=2.52 Hz, 1H); Tr (3 min)=1.97 min m/z (ES+) no ionization.

(152) Referring to Reaction Scheme 22, Stage 2. 4-Bromo-2-chloro-6-nitrophenol (12.0 g, 47.5 mmol) was dissolved in ethanol (350 mL). Water (80 mL) and saturated aqueous ammonium chloride (80 mL) were added, followed by iron powder (21.2 g, 380 mmol). The reaction was heated to 50? C. and stirred for 2 h. The reaction was cooled to rt and filtered through a prewashed pad of celite, washing with 100 mL EtOH followed by excess EtOAc (approx 1.5 l) until the liquid ran clear. The filtrate was concentrated to remove organic solvents. EtOAc (approx 400 mL) was added to the aqueous residue and the layers were separated. The organic phase was washed with water (150 mL) and brine (100 mL). The aqueous layers were re-extracted with EtOAc (2?150 mL). The combined organics were filtered to remove a pale brown solid and evaporated to dryness to give a purple solid. Dry flash chromatography (Elution: 0-30% EtOAc-heptane) afforded the desired compound (6.5 g, 61% yield) as a pale solid. ?H (500 MHz, DMSO) 9.01 (br. s., 1H) 6.71 (d, J=2.36 Hz, 1H) 6.66 (d, J=2.36 Hz, 1H) 5.23 (br. s., 2H); Tr (3 min)=1.70 min m/z (ES+) (M+H)+222, 224, 226.

(153) Referring to Reaction Scheme 22, Stage 3. 2-Amino-4-bromo-6-chlorophenol (2.04 g, 9.18 mmol) was dissolved in DCM (anhydrous, 30 ml). Triethylamine (1.6 ml, 11.5 mmol) was added and the reaction was stirred at rt for 1 h under nitrogen. The reaction was cooled in an ice bath for 15 min and then cyclopropanecarbonyl chloride (0.700 mL, 7.65 mmol) was added dropwise over a period of 20 min. The reaction was allowed to gradually warm to rt and stirred for 2 h at rt. The reaction was cooled in an ice bath and an extra 0.2 eq. acid chloride was added dropwise. The reaction was allowed to warm to rt and stirred at rt for 2 h. DCM (20 mL) was added to the reaction followed by water (50 mL). The organic and aqueous layers were separated. The organic layer was washed with water (3?50 mL), brine (30 mL), dried (MgSO4), filtered and concentrated to give the desired product which was carried forward without further purification.

(154) Referring to Reaction Scheme 22, Stage 4. A crude 4:1:1 mixture of N-(5-bromo-3-chloro-2-hydroxyphenyl)cyclopropanecarboxamide, 2-amino-4-bromo-6-chlorophenylcyclopropanecarboxylate and 4-bromo-2-chloro-6-cyclopropaneamido

(155) phenylcyclopropanecarboxylate (2.77 g) was dissolved in toluene (30 mL). TsOH monohydrate (2.54 g, 13.4 mmol) was added and the reaction was stirred at 115? C. for 16 h. The reaction was cooled to rt and concentrated to give a brown oil. The residue was re-dissolved in EtOAc (100 mL). The solution was washed with saturated aqueous sodium bicarbonate (3?100 mL), water (3?100 mL), brine (50 mL) and dried (MgSO4). Filtration and concentration gave a brown oil. Column chromatography (Elution: 0-10% EtOAc-heptane) afforded the desired compound (1.18 g, 42%) as an orange crystalline solid. ?H (500 MHz, DMSO) 7.85 (d, J=1.73 Hz, 1H) 7.68 (d, J=1.58 Hz, 1H) 2.27-2.40 (m, 1H) 1.08-1.38 (m, 4H); Tr (3 min)=2.38 min m/z (ES+) (M+H)+272, 274.

(156) Referring to Reaction Scheme 22, Stages 5, 6 & 7 were carried out as described in Reaction Scheme 15.

(157) The following compounds were prepared substantially as described above.

(158) TABLE-US-00022 Mole- Mass cular Spec Structure Weight Result 315.72 [M + H]+ = 316/318, 100% @ rt = 3.84 min

Example 23

(159) ##STR00097##

(160) Referring to Reaction Scheme 23, Stage 1. 2-amino-4-bromo-6-chlorophenol (2.50 g, 11.2 mmol) was dissolved in THF (30 ml). CDI (2.73 g, 16.9 mmol) was added and the reaction was stirred at 65? C. After 2 h the reaction was cooled to rt and concentrated to give an orange solid. The residue was redissolved in EtOAc (100 mL) and the organic phase was washed with water (50 mL), 2M HCl (3?50 mL), water (100 mL) and brine (20 mL) and dried (MgSO4). Filtration and concentration afforded the desired compound (2.7 g, 97% yield) as a white solid. ?H (500 MHz, DMSO-d6) 12.01 (br. s., 1H) 7.44 (d, J=1.73 Hz, 1H) 7.26 (d, J=1.73 Hz, 1H); Tr (3 min)=1.87 min m/z (ES?) (M?H)? 246, 248.

(161) Referring to Reaction Scheme 23, Stage 2. 5-Bromo-7-chloro-2,3-dihydro-1,3-benzoxazol-2-one (0.60 g, 2.4 mmol) was dissolved in anhydrous DMF (10 mL) and the reaction was cooled in an ice bath. Sodium hydride (60% in oil, 0.15 g, 3.6 mmol) was added portionwise and the reaction was stirred in the ice bath for 1 h. Methyl iodide (0.18 ml, 0.29 mmol) was added and the reaction was stirred at rt for 2 hours. The reaction was cooled in a slush bath. Water (5 mL) was added cautiously followed by EtOAc (20 mL). The layers were separated. The aqueous was re-extracted with EtOAc (2?15 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL) and dried (MgSO4). Filtration and concentration gave a colourless oil. Column chromatography (Elution: 0-20% EtOAc-heptane) afforded the desired compound (540 mg, 85% yield) as a pink solid. ?H (500 MHz, CDCl3) 7.30 (d, J=1.73 Hz, 1H) 7.03 (d, J=1.73 Hz, 1H) 3.41 (s, 3H); Tr (3 min)=1.97 min m/z (ES+) No ionisation.

(162) Referring to Reaction Scheme 23, Stages 3, 4 & 5 were carried out as described in Reaction Scheme 15.

(163) The following compounds were prepared substantially as described above.

(164) TABLE-US-00023 Mole- cular Mass Spec Structure Weight Result 305.68 [M + H]+ = 306/308, 98% @ rt = 3.35 min

Example 24

(165) ##STR00099##

(166) Referring to Reaction Scheme 24, Stage 1. Methylmagnesium bromide (1.4M in toluene/THF, 1.5 mL, 0.046 mol) was added drop wise over 1 hour to a cold (?78? C.), stirred solution of 4-bromo-2-chlorobenzaldehyde (5.0 g, 0.023 mol) in THF (100 mL) and the mixture was stirred at this temperature under a nitrogen atmosphere for 1 hour. After this time, the reaction mixture was allowed to warm to room temperature over 1 hour before being stirred for a further 1.5 hours. The reaction mixture was then cooled to 5? C. in an ice bath and stirred for 10 minutes before saturated ammonium chloride (40 mL) was added drop wise and stirring continued at this temperature for a further 10 minutes before being allowed to warm to room temperature. The resulting mixture was then extracted with ethyl acetate (1?100 mL), the organic layer was washed sequentially with water (100 mL), and brine (100 mL) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 10% ethyl acetate, 90% heptanes) to give the desired compound (4.33 g, 81% yield) as a colourless oil. 6H (500 MHz, DMSO) 7.64 (d, J=1.58 Hz, 1H) 7.49-7.60 (m, 2H) 5.47 (d, J=3.00 Hz, 1H) 4.96 (dd, J=6.07, 2.60 Hz, 1H) 1.28 (d, J=6.31 Hz, 3H).

(167) Referring to Reaction Scheme 24, Stage 2. Sodium hydride (60% in oil, 0.38 g, 9.6 mmol) was added portion wise over 5 minutes to a cooled (0? C.), stirred solution of 1-(4-bromo-2-chlorophenyl)ethan-1-ol (1.5 g, 6.4 mmol) in DMF (15 mL) and the reaction was stirred at this temperature for 20 minutes under a nitrogen atmosphere. After this time, methyl iodide (0.48 mL, 7.6 mmol) was added in one portion and the reaction mixture was allowed to warm to room temperature before being stirred for a further 18 hours. The reaction was quenched by the drop wise addition of water (15 mL) over 10 minutes and the resulting solution was extracted with ethyl acetate (2?30 mL). The combined organic extracts were washed sequentially with water (100 mL) and brine (10 mL) before being dried (MgSO4), filtered and concentrated to give the desired compound (1.5 g, 99% yield) as a yellow oil. 6H (500 MHz, DMSO) 7.71 (d, J=1.89 Hz, 1H) 7.60 (dd, J=8.35, 1.89 Hz, 1H) 7.39 (d, J=8.35 Hz, 1H) 4.63 (q, J=6.46 Hz, 1H) 3.16 (s, 3H) 1.26-1.38 (m, 3H).

(168) Referring to Reaction Scheme 24, Stages 3, 4 & 5 were carried out as described in Reaction Scheme 15.

(169) The following compounds were prepared substantially as described above.

(170) TABLE-US-00024 Mole- Mass cular Spec Structure Weight Result 00 292.72 [M + H]+ = 293/295, 99% @ rt = 3.72 min

Example 25

(171) ##STR00101##

(172) Referring to Reaction Scheme 25, Stage 1. [(1-Ethoxycyclopropyl)oxy](trimethyl)silane (5.6 mL, 27.85 mmol) was added drop wise over 10 minutes to a stirred solution of 4-bromo-2-chloroaniline (5.0 g, 24.22 mmol) in a mixture of methanol (50 mL) and acetic acid (95 mL) and the resulting solution was heated to 70? C. and stirred at this temperature for 4 hours. After this time, the reaction mixture was cooled to room temperature and concentrated. The resulting residue was then dissolved in THF (25 mL) and added drop wise to a cooled (0? C.), stirred solution of sodium borohydride (1.87 g, 49.4 mmol) and (diethyl ether)(trifluoro)boron (6.2 mL, 48.9 mmol) in THF (50 mL). The resulting mixture was then heated to 70? C. and stirred at this temperature for 4 hours before being cooled to room temperature and allowed to stand overnight. The resulting reaction mixture was quenched by the addition of water (100 mL) before being extracted with ethyl acetate (3?30 mL). The combined organic extracts were washed sequentially with water (100 mL) and brine (100 mL) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified on a Biotage isolera (5% ethyl acetate, 95% heptanes) to give the desired compound (4.8 g, 76% yield) as a colourless oil. Tr=2.44 min m/z (ES+) (M+H+) 246/248.

(173) Referring to Reaction Scheme 25, Stage 2. Sodium hydride (60% dispersion in oil, 0.29 g, 7.28 mmol) was added in one portion to a cooled (0? C.) stirred solution of 4-bromo-2-chloro-N-cyclopropylaniline (1.4 g, 5.68 mmol) in dry DMF (35 mL) and the resulting solution was stirred for 5 minutes. After this time, iodomethane (0.35 mL, 5.62 mmol) was added and the reaction mixture was stirred for 10 minutes before being allowed to warm to room temperature and stirred for a further 6 hours under a nitrogen atmosphere. The resulting reaction mixture was extracted with ethyl acetate (3?25 mL) and the organic layer washed sequentially with water (75 mL) and brine (75 ml) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by dry flash chromatography (elution: 100% heptanes) to give the desired compound (1.44 g, 78% yield) as a colourless oil. 6H (500 MHz, DMSO) 7.56 (d, J=2.36 Hz, 1H), 7.46 (dd, J=8.67, 2.36 Hz, 1H), 7.31 (d, J=8.67 Hz, 1H), 2.81 (s, 3H), 2.53-2.58 (m, 1H), 0.63-0.69 (m, 2H), 0.27-0.33 (m, 2H).

(174) Referring to Reaction Scheme 25, Stages 3, 4 & 5 were carried out as described in Reaction Scheme 15.

(175) The following compounds were prepared substantially as described above.

(176) TABLE-US-00025 Mole- Mass cular Spec Structure Weight Result 02 289.72 [M + H]+ = 290/292, 98% @ rt = 3.77 min 03 303.75 [M + H]+ = 304/306, 100% @ rt = 4.40 min

Example 26

(177) ##STR00104##

(178) Referring to Reaction Scheme 26, Stage 1. Bromine (0.54 mL, 10.4 mmol) was added drop wise to a cooled (0? C.), stirred solution of 2-amino-3-chlorophenol (1.0 g, 6.97 mmol) in DCM (50 mL) and the resulting solution was warmed to room temperature and stirred for 16 hours. After this time, the reaction mixture was cooled in an ice-bath and bromine (0.11 mL, 2.09 mmol) was added before being warmed to room temperature and stirred for a further 1 hour. The resulting solid precipitate was collected by filtration, suspended in DCM (100 mL) and washed with saturated sodium bicarbonate (50 mL). The organic layer was removed, washed sequentially with water (10 mL) and brine (10 mL), before being dried (MgSO4), filtered and concentrated to give the desired compound (1.0 g, 64% yield) as a red solid. ?H (500 MHz, DMSO) 10.13 (br. s., 1H), 6.89 (d, J=2.21 Hz, 1H), 6.75 (d, J=2.21 Hz, 1H), 4.82 (br. s., 2H).

(179) Referring to Reaction Scheme 26, Stage 2. p-Toluene sulfonic acid (0.02 g, 0.12 mmol) was added in one portion to a stirred solution of 2-amino-5-bromo-3-chlorophenol (0.9 g, 4.05 mmol) in triethylorthoacetate (10 mL) and the resulting reaction mixture was heated to 140? C. and stirred at this temperature for 18 hours. After this time, the reaction mixture was cooled to room temperature and partitioned between water (10 mL) and ethyl acetate (20 mL). The organic layer was removed, washed sequentially with water (10 mL), saturated sodium bicarbonate (2?20 mL) and brine (10 mL) before being dried (MgSO4), filtered and concentrated. The resulting residue was purified on a Biotage isolera (0% ethyl acetate, 100% heptanes to 40% ethyl acetate, 60% heptanes) to give the desired compound (0.68 g, 48% yield) as a red solid. ?H (500 MHz, CDCl3) 7.58 (d, J=1.42 Hz, 1H), 7.50 (d, J=1.58 Hz, 1H), 2.61-2.73 (m, 3H).

(180) Referring to Reaction Scheme 26, Stages 3, 4 & 5 were carried out as described in Reaction Scheme 15.

(181) The following compounds were prepared substantially as described above.

(182) TABLE-US-00026 Mole- Mass cular Spec Structure Weight Result 05 289.67 [M + H]+ = 290/292 100% @ rt = 3.26 min

Example 27

(183) The following compounds may be prepared substantially as described above.

(184) TABLE-US-00027 06 6-(3-chloro-4-{[1- (morpholin-4-yl)propan-2- yl]oxy}phenyl) pyrimidine-4-carboxylic acid 07 6-[3-chloro-4- (cyclopropoxymethyl) phenyl]pyrimidine- 4-carboxylic acid 08 6-[3-chloro-4- (cyclopropylmethyl)phenyl] pyrimidine-4- carboxylic acid 09 6-[3-chloro-4- (cyclopropylsulfanyl)phenyl] pyrimidine- 4-carboxylic acid 0 6-[3-chloro-4- (cyclopropylsulfinyl)phenyl] pyrimidine- 4-carboxylic acid 6-[3-chloro-4- (cyclopropanesulfonyl)phenyl] pyrimidine- 4-carboxylic acid 6-{3-chloro-4- [cyclopropyl(hydroxy) methyl]phenyl}pyrimidine-4- carboxylic acid 6-[3-chloro-4-(1- cyclopropoxyethyl) phenyl]pyrimidine- 4-carboxylic acid 6-(3-chloro-4-cyclopropane- carbonylphenyl)pyrimidine- 4-carboxylic acid 6-(3-chloro-4- cyclopropylphenyl)pyrimidine-4- carboxylic acid 6-[4-(aziridin-1-ylmethyl)-3- chlorophenyl]pyrimidine-4- carboxylic acid 6-{3-chloro-4- [(dimethylamino)methyl] phenyl}pyrimidine-4- carboxylic acid 6-[3-chloro-4-(cyclopropylamino) phenyl]pyrimidine-4- carboxylic acid 6-{3-chloro-4- [cyclopropyl(methyl)amino] phenyl}pyrimidine-4- carboxylic acid 0 6-{3-chloro-4- [(cyclopropylamino)methyl] phenyl}pyrimidine-4- carboxylic acid 6-(3-chloro-4- {[cyclopropyl(methyl)amino] methyl}phenyl)pyrimidine- 4-carboxylic acid 6-(7-chloro-2-cyclopropyl- 2,3-dihydro-1H-isoindol-5- yl)pyrimidine-4-carboxylic acid 6-[3-chloro-4-(furan-2-yl) phenyl]pyrimidine-4- carboxylic acid 6-[3-chloro-4-(1- methoxycyclopropyl)phenyl] pyrimidine-4-carboxylic acid 6-(2,3-dihydro-1,4- benzodioxin-6-yl) pyrimidine-4- carboxylic acid 6-(7-chloro-2-methyl-1,3- benzoxazol-5-yl) pyrimidine-4- carboxylic acid 6-(7-chloro-2-oxo-2,3- dihydro-1,3-benzoxazol-5- yl)pyrimidine- 4-carboxylic acid 6-(7-chloro-3-methyl- 2-oxo-2,3-dihydro-1,3- benzoxazol-5-yl)pyrimidine- 4-carboxylic acid 6-(7-chloro-2-cyclopropyl- 1,3-benzoxazol-5- yl)pyrimidine-4- carboxylic acid 0 6-{8-chloroimidazo[1,2-a] pyridin-6-yl}pyrimidine-4- carboxylic acid 6-(4-chloro-1,3-benzoxazol- 6-yl)pyrimidine-4- carboxylic acid 6-(quinolin-6-yl)pyrimidine- 4-carboxylic acid 6-{pyrazolo[1,5-a]pyridin-5-yl} pyrimidine-4-carboxylic acid 6-(4-chloro- 3-cyclopropoxyphenyl) pyrimidine-4- carboxylic acid 6-(4-chloro-3- methoxyphenyl)pyrimidine- 4-carboxylic acid 6-[4-chloro-3-(propan- 2-yloxy)phenyl]pyrimidine-4- carboxylic acid 6-[4-chloro-3-(2- methylpropoxy)phenyl] pyrimidine-4- carboxylic acid 6-[4-chloro-3- (trifluoromethoxy)phenyl] pyrimidine-4- carboxylic acid 6-{4-chloro-3- [(1,1,1-trifluoropropan-2- yl)oxy]phenyl}pyrimidine- 4-carboxylic acid 0 6-(benzo[d][1,3]dioxol-5- yl)pyrimidine-4-carboxylic acid 6-(2,2-difluorobenzo[d] [1,3]dioxol-5-yl)pyrimidine-4- carboxylic acid 6-(2,3-dihydrobenzo[b] [1,4]dioxin-6-yl)pyrimidine-4- carboxylic acid 6-(7-chlorobenzo[b] thiophen-5-yl)pyrimidine-4- carboxylic acid 6-(7-chlorobenzo[d]thiazol- 5-yl)pyrimidine-4-carboxylic acid 6-(7-chlorobenzo[d]oxazol- 5-yl)pyrimidine-4-carboxylic acid 6-(7-chlorobenzo[c][1,2,5] oxadiazol-5-yl)pyrimidine-4- carboxylic acid 6-(7-chloro-2,3,3a,7a- tetrahydrobenzofuran-5- yl)pyrimidine-4-carboxylic acid 6-(7-chloro-3a,7a-dihydro- 1H-indol-5-yl)pyrimidine-4- carboxylic acid 6-(7-chloro-1-methyl-3a, 7a-dihydro-1H-indazol-5- yl)pyrimidine-4-carboxylic acid 0 6-(8-chloroquinazolin-6-yl) pyrimidine-4-carboxylic acid 6-(5-chloroquinazolin-7-yl) pyrimidine-4-carboxylic acid 6-(8-chloroquinoxalin-6-yl) pyrimidine-4-carboxylic acid 6-(7-chloro-1H-benzo[d] imidazol-5-yl)pyrimidine-4- carboxylic acid 6-(3-chloro-4- (1-methylcyclopropyl) phenyl)pyrimidine- 4-carboxylic acid 6-(3-chloro-4-(1- (trifluoromethyl)cyclopropyl) phenyl)pyrimidine-4- carboxylic acid 6-(3-chloro-4-(3-methyloxetan- 3-yl)phenyl)pyrimidine- 4-carboxylic acid 6-(3-chloro-4-(pyrrolidin-1- yl)phenyl)pyrimidine-4- carboxylic acid 6-(3-chloro-4-(pyrrolidin-3- yl)phenyl)pyrimidine-4- carboxylic acid 6-(3-chloro-4-(pyrrolidin-2- yl)phenyl)pyrimidine-4- carboxylic acid 0 6-(3-chloro-4-(1H-imidazol-2- yl)phenyl)pyrimidine-4- carboxylic acid 6-(3-chloro-4-(1H-pyrrol-2- yl)phenyl)pyrimidine-4- carboxylic acid 6-(4-tert-butyl-3-chlorophenyl) pyrimidine-4-carboxylic acid 7-chloro-8-cyclopropoxy- 5H-chromeno[4,3- d]pyimidine-4-carboxylic acid

Example 28

(185) A generalized procedure for monitoring L-Kynurenine (KYN) hydroxylation to form product 3-Hydroxy-Kynurenine (30H-KYN) by LC/MS is described below. Product is quantified by multiple reaction monitoring using MS.

(186) Key Reagents:

(187) Compound: Stock concentrations: 10 mM in 100% DMSO Cell line: CHO GST HIS KMO cell line, 1E4 cells/well/100 ?l in 96 well cell plate Substrate: L-Kynurenine (Sigma: Cat# K3750, stock concentration: 10 mM in 100 mM potassium phosphate buffer, pH 7.4)
Assay Conditions: Medium: OptiMem (Reduced Serum Medium 1?, +L-Glutamine+HEPESPhenol Red; GIBCO: Cat#11058) Assay Volume: 200 ?l Plate Format: 96 well plate, transparent (Corning) Read-Out: product (30H-KYN) quantification using product specific MRM Reader: LC/MS/MS
Assay Protocol: prepare serial dilution (factor 3) of compound in 100% DMSO (top concentration=6.67 mM, 100% DMSO) [8 points: 6.67 mM; 2.22 mM; 0.74 mM; 0.247 mM; 0.082 mM; 0.027 mM; 0.009 mM; 0.003 mM] prepare 300-fold concentrated solution of each compound concentration (top concentration 22.22 ?M, 0.3% DMSO) in OptiMem medium [22.2 ?M; 7.41 ?M; 2.47 ?M; 0.82 ?M; 0.27 ?M; 0.09 ?M; 0.03 ?M; 0.041M] prepare substrate (10 mM) at concentration of 1.1 mM in medium medium of cell plate is drawed off cells are washed with OptiMem (100 ?l/well) and drawed off again assay mix: 90 ?l OptiMem/well+90 ?l compound/well of each concentration [final compound top concentration: 10 ?M; 0.15% DMSO] [final compound bottom concentration: 0.004 ?M; 0.15% DMSO] pre-incubation: 30 min at 37? C. add 20 ?l/well of the 1.1 mM substrate solution (final assay concentration: 100 ?M) positive control: 200 ?l OptiMem negative control: 180 ?l OptiMem+20 ?l 1.1 mM substrate incubate ?24 h at 37? C. transfer 100 ?l of each well in a transparent 96 well plate (Corning) add 100 ?l/well 10% trichloro acetic acid (TCA) in water centrifugate plate for 3 min at 4000 rpm detect product by LC/MS (injection of 50 ?l/well; 2.5 fold overfill of the 20 ?l sample loop)
Data Analysis:
IC.sub.50's are calculated using automated fitting algorithm (A+Analysis).

Example 29

(188) A method of monitoring L-Kynurenine (KYN) hydroxylation to form product 3-Hydroxy-Kynurenine (30H-KYN) by LC/MS is described below. Product is quantified by multiple reaction monitoring.

(189) Key Reagents:

(190) Compound: Stock concentrations: 10 mM in 100% DMSO Enzyme: KMO enzyme prepared at Evotec via mitochondria isolation from CHO-GST HIS KMO cells Substrate: L-Kynurenine (Sigma: Cat# K3750) [stock concentration: 10 mM in 100 mM potassium phosphate buffer, pH 7.4]
Assay Conditions: Buffer: 100 mM potassium phosphate, pH 7.4, 200 ?M NADPH, 0.4 U/ml G6P-DH (Glucose 6-phosphate dehydrogenase), 3 mM G6P (D-Glucose 6-phosphate) Assay Volume: 40 ?l Plate Format: 384 well plate, transparent (Matrix) Read-Out: product (30H-KYN) quantification using product specific MRM Reader: LC/MS/MS
Assay Protocol: prepare serial dilution (factor 3) of compound in 100% DMSO (top concentration=10 mM, 100% DMSO) [8 points: 10 mM; 3.33 mM; 1.11 mM; 0.37 mM; 0.12 mM; 0.04 mM; 0.0137 mM; 0.0045 mM, 0.0015 mM] prepare 3.33-fold concentrated solution of each compound concentration (top concentration 300 ?M, 3% DMSO) in assay buffer [concentrations: 300 ?M; 100 ?M; 33.3 ?M; 11.1 ?M; 3.70 ?M; 1.23 ?M; 0.41 ?M; 0.137 ?M] prepare substrate (10 mM) at concentration of 1 mM in assay buffer assay mix: 4 ?l compound/well of each concentration+24 ?l assay buffer/well+8 ?l KMO human enzyme+4 ?l 1 mM substrate (final concentration=100 ?M) [final compound top concentration: 30 ?M; 0.3% DMSO] [final compound bottom concentration: 0.0137 ?M; 0.3% DMSO] positive control: 4 ?l 50 ?M FCE28833 in assay buffer [0.5% DMSO] (final assay concentration=5 ?M)+24 ?l assay buffer/well+8 ?l KMO human enzyme+4 ?l 1 mM substrate (final concentration=100 ?M) negative control: 28 ?l assay buffer/well+8 ?l KMO human enzyme+4 ?l 1 mM substrate (final concentration=100 ?M) incubate 400 min at RT add 40 ?l/well 10% trichloro acetic acid in water to stop the assay and precipitate protein centrifuge plate for 3 min at 4000 rpm product detection by LC/MS (injection of 50 ?l/well; 2.5 fold overfill of the 20 ?l sample loop)
Data Analysis:
IC.sub.50's are calculated using automated fitting algorithm (A+Analysis).

Example 30

(191) A method of monitoring L-Kynurenine (KYN) hydroxylation to form 3-Hydroxy-Kynurenine (30H-KYN) by LC/MS is described. Product is quantified by multiple reaction monitoring (MRM method).

(192) Key Reagents:

(193) Compound: Stock concentrations: 10 mM in 100% DMSO Enzyme: KMO enzyme prepared at Evotec from mouse liver (4-6 weeks old) via mitochondria isolation as described in the literature Substrate: L-Kynurenine (Sigma: Cat# K3750, stock concentration: 10 mM in 100 mM potassium phosphate buffer, pH 7.4)
Assay Conditions: Buffer: 100 mM potassium phosphate, pH 7.4, 200 ?M NADPH, 0.4 U/ml G6P-DH (Glucose 6-phosphate Dehydrogenase), 3 mM G6P (D-Glucose 6-phosphate) Assay Volume: 40 ?l Plate Format: 384 well plate, transparent (Matrix) Read-Out: product (3OH-KYN) quantification using product specific MRM Reader: LC/MS/MS
Assay Protocol: prepare serial dilution (factor 3) of compound in 100% DMSO (top concentration=10 mM, 100% DMSO) [8 points: 10 mM; 3.33 mM; 1.11 mM; 0.37 mM; 0.12 mM; 0.04 mM; 0.0137 mM; 0.0045 mM, 0.0015 mM] prepare 3.33-fold concentrated solution of each compound concentration (top concentration 300 ?M, 3% DMSO) in assay buffer [concentrations: 300 ?M; 100 ?M; 33.3 ?M; 11.1 ?M; 3.70 ?M; 1.23 ?M; 0.41 ?M; 0.137 ?M] prepare substrate (10 mM) at concentration of 1 mM in assay buffer assay mix: 4 ?l compound/well of each concentration+24 ?l assay buffer/well+8 ?l KMO mouse enzyme+4 ?l 1 mM substrate (final concentration=100 ?M) [final compound top concentration: 30 ?M; 0.3% DMSO] [final compound bottom concentration: 0.0137 ?M; 0.3% DMSO] positive control: 4 ?l 50 ?M FCE28833 in assay buffer, 0.5% DMSO [final assay concentration=5 ?M]+24 ?l assay buffer/well+8 ?l KMO mouse enzyme+4 ?l 1 mM substrate [final concentration=100 ?M] negative control: 28 ?l assay buffer/well+8 ?l KMO mouse enzyme+4 ?l 1 mM substrate [final concentration=100 ?M] incubate 40 min at RT add 40 ?l/well 10% trichloro acetic acid in water to stop the assay and precipitate protein centrifuge plate for 3 min at 4000 rpm product detection by LC/MS (injection of 20 ?l/well, 2 fold overfill of the 10 ?l sample loop)
Data Analysis:
IC.sub.50's are calculated using automated fitting algorithm (A+Analysis).

Example 31

(194) Using procedures similar to those described herein, the following compounds were assayed for activity.

(195) TABLE-US-00028 % Inhibition IUPAC name at 10 uM* 6-(4-Chloro-3-methoxy-phenyl)-pyrimidine-4-carboxylic 99.62 acid 6-(3-Amino-4-chloro-phenyl)-pyrimidine-4-carboxylic 101.01 acid 6-[4-Chloro-3-(tetrahydro-furan-3-yloxy)-phenyl]- 88.39 pyrimidine-4-carboxylic acid pyridin-3-ylamide 6-[4-Chloro-3-(2-morpholin-4-yl-ethoxy)-phenyl]- 61.41 pyrimidine-4-carboxylic acid hydrochloride salt 6-(3-Chloro-4-isopropyl-phenyl)-pyrimidine-4-carboxylic 100 acid 6-(3-Fluoro-4-methyl-phenyl)-pyrimidine-4-carboxylic 100 acid 6-(3-Chloro-4-isopropoxy-phenyl)-pyrimidine-4- 100 carboxylic acid 6-(3-Chloro-4-isopropoxy-phenyl)-2-methyl-pyrimidine- 70 4-carboxylic acid 6-(3-Fluoro-4-methyl-phenyl)-2-methyl-pyrimidine-4- 96 carboxylic acid 6-(3-Chloro-4-cyclopentyloxy-phenyl)-pyrimidine-4- 97 carboxylic acid 6-(3-Chloro-4-trifluoromethoxy-phenyl)-pyrimidine-4- 100 carboxylic acid 6-(3-Fluoro-4-isopropyl-phenyl)-pyrimidine-4-carboxylic 85 acid 6-(4-(R)-sec-Butoxy-3-chloro-phenyl)-pyrimidine-4- 100 carboxylic acid 6-(4-(S)-sec-Butoxy-3-chloro-phenyl)-pyrimidine-4- 100 carboxylic acid 6-(3-Chloro-4-cyclopropoxy-phenyl)-pyrimidine-4- 100 carboxylic acid 6-[3-Chloro-4-(2,2,2-trifluoro-1-methyl-ethoxy)-phenyl]- 94 pyrimidine-4-carboxylic acid 4-(3-Chloro-4-cyclopropoxy-phenyl)-pyridine-2- 100 carboxylic acid 6-(4-(R)-sec-Butoxy-3-chloro-phenyl)-pyridine-4- 50 carboxylic acid 6-(4-(S)-sec-Butoxy-3-chloro-phenyl)-pyridine-4- 82 carboxylic acid 4-(3-Chloro-4-isopropoxy-phenyl)-pyridine-2-carboxylic 80 acid 4-(3-Chloro-4-trifluoromethoxy-phenyl)-pyridine-2- 89 carboxylic acid 6-(3-Chloro-4-cyclobutoxy-phenyl)-pyrimidine-4- 100 carboxylic acid 6-[3-Chloro-4-(2-piperidin-1-yl-ethoxy)-phenyl]- 90 pyrimidine-4-carboxylic acid 6-Quinolin-6-yl-pyrimidine-4-carboxylic acid 100 6-(8-Chloro-chroman-6-yl)-pyrimidine-4-carboxylic acid 100 6-(7-Chloro-benzofuran-5-yl)-pyrimidine-4-carboxylic 100 acid 6-[3-Chloro-4-(pyrrolidin-3-yloxy)-phenyl]-pyrimidine-4- 80 carboxylic acid 6-(8-chloro-1-methyl-1,2,3,4-tetrahydroquinolin-6- 100 yl)pyrimidine-4-carboxylic acid 6-(8-chloroquinolin-6-yl)pyrimidine-4-carboxylate 100 N-[6-(3-chloro-4-cyclopropoxyphenyl)pyrimidin-4- 73 yl]benzenesulfonamide N-[6-(3-chloro-4-cyclopropoxyphenyl)pyrimidin-4-yl]-4- 98 fluorobenzene-1-sulfonamide N-[6-(3-chloro-4-cyclopropoxyphenyl)pyrimidin-4-yl]-4- 88 (trifluoromethoxy)benzene-1-sulfonamide N-[6-(3-chloro-4-cyclopropoxyphenyl)pyrimidin-4-yl]-3- 77 (trifluoromethoxy)benzene-1-sulfonamide N-[6-(3-chloro-4-cyclopropoxyphenyl)pyrimidin-4-yl]-2- 96 fluorobenzene-1-sulfonamide N-[6-(3-chloro-4-cyclopropoxyphenyl)pyrimidin-4- 33 yl]cyclopropanesulfonamide 6-(8-chloro-1,2,3,4-tetrahydroquinolin-6-yl)pyrimidine-4- 100 carboxylate 6-(3-chloro-4-cyclopropoxyphenyl)-5-methylpyrimidine- 100 4-carboxylate 6-{3-chloro-4-[2-(morpholin-4- 99 yl)ethoxy]phenyl}pyrimidine-4-carboxylate 6-[3-chloro-4-(cyclopropylmethoxy)phenyl]pyrimidine-4- 101 carboxylate 6-[3-chloro-4-(oxetan-3-yloxy)phenyl]pyrimidine-4- 100 carboxylate 4-(3-chloro-4-cyclopropoxyphenyl)-5H,7H-furo[3,4- 100 d]pyrimidin-7-one 6-(3-chloro-4-cyclopropoxyphenyl)-5- 100 (hydroxymethyl)pyrimidine-4-carboxylic acid 4-(3-chloro-4-cyclopropoxyphenyl)-5H,6H,8H- 100 pyrano[3,4-d]pyrimidin-8-one [(2R,3S,4S,5R)-3,4,5,6-tetrahydroxyoxan-2-yl]methyl 6- 102 (3-chloro-4-cyclopropoxyphenyl)pyrimidine-4-carboxylate 6-[3-chloro-4-(methylsulfanyl)phenyl]pyrimidine-4- 103 carboxylic acid 6-[3-chloro-4-(methylsulfinyl)phenyl]pyrimidine-4- 100 carboxylic acid 6-[3-chloro-4-(methylsulfonyl)phenyl]pyrimidine-4- 100 carboxylic acid 6-{3-chloro-4- 90 [cyclopropyl(hydroxy)methyl]phenyl}pyrimidine-4- carboxylic acid 6-(3-chloro-4-cyclopropanecarbonylphenyl)pyrimidine-4- 101 carboxylic acid 6-[3-chloro-4-(methoxymethyl)phenyl]pyrimidine-4- 105 carboxylic acid 6-[3-chloro-4-(1-methoxyethyl)phenyl]pyrimidine-4- 101 carboxylic acid 6-{3-chloro-4- 65 [(dimethylamino)methyl]phenyl}pyrimidine-4-carboxylic acid 6-[3-chloro-4-(cyclopropylamino)phenyl]pyrimidine-4- 101 carboxylic acid 6-{3-chloro-4- 96 [cyclopropyl(methyl)amino]phenyl}pyrimidine-4- carboxylic acid 6-(3-chloro-4-(pyrrolidin-1-yl)phenyl)pyrimidine-4- 100 carboxylic acid 6-(7-chloro-2-methyl-1,3-benzoxazol-5-yl)pyrimidine-4- 102 carboxylic acid 6-(8-chloroquinoxalin-6-yl)pyrimidine-4-carboxylic acid 102 6-(7-chloro-2,3-dihydro-1-benzofuran-5-yl)pyrimidine-4- 102 carboxylic acid 6-(7-chloro-2-cyclopropyl-1,3-benzoxazol-5- 100 yl)pyrimidine-4-carboxylic acid 6-(4-chloro-2-methyl-1,3-benzoxazol-6-yl)pyrimidine-4- 102 carboxylic acid 6-(7-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazol- 100 5-yl)pyrimidine-4-carboxylic acid 6-(2H-1,3-benzodioxol-5-yl)pyrimidine-4-carboxylic acid 101 *Some portion of activity of amides may be due to contribution of acid precursor.

Example 32: General Procedures

(196) Method A.

(197) Amide coupling. To a solution of carboxylic acid (1 eq) in DMF were added EDC.HCl (1 eq) and HOBt (1 to 1.2 eq) or HATU (1 to 1.2 eq). The reaction mixture was stirred at ambient temperature for 30 minutes after which time the appropriate amine (1 eq) was added. The reaction was monitored by LCMS to completion whereupon the reaction mixture was poured into water. The resultant precipitate was filtered, washed with water (?2), heptane (?2) and dried in vacuo to yield the target compound. If a precipitate was not formed the reaction mixture was extracted with EtOAc (?3) and the combined organic layers were washed with water (?2), saturated aqueous NaCl (?2), dried (Na2SO4 or MgSO4) and the solvent removed in vacuo to afford the crude product. Purification was carried out by flash column chromatography, prep HPLC, or a combination of both.

(198) Method B.

(199) Amide coupling. To a solution of carboxylic acid (1 eq) in DCM (20 vol) under nitrogen were added oxalyl chloride (3 eq) and 1 drop of DMF (cat.). The reaction mixture was stirred at ambient temperature for 30 minutes after which time the solvents were removed in vacuo. DCM (20 vol) or THF (20 vol) was added, followed by the required amine (1 to 3 eq) and triethylamine (2 eq) or DIPEA (1.5 eq). The reaction mixture was stirred at ambient temperature. The reaction was monitored by LCMS to completion whereupon water was added. The reaction mixture was then extracted with DCM and the organic layer was washed with water, saturated aqueous NaCl, dried over Na2SO4 or MgSO4 and the solvent removed in vacuo to afford the crude product. Purification was carried out by flash column chromatography, prep HPLC, a combination of both or by trituration with an appropriate solvent.

(200) Method C.

(201) Amide coupling. To a solution of carboxylic acid (1 eq) in DMF were added EDC.HCl (1 eq) and HOBt (1 eq). The reaction mixture was stirred at ambient temperature for 30 minutes after which time the appropriate amine was added. The reaction was monitored by LCMS. After completion the reaction mixture was poured into water after which a precipitate came out of solution and was filtered, washed with water, heptane and dried in vacuo to yield the target compound or if a precipitate was not formed the reaction mixture was extracted with EtOAc (3?) and the combined organic layers were washed with water, saturated aqueous NaCl, dried (Na2SO4 or MgSO4) and the solvent removed in vacuo to afford the crude product. Purification was carried out by flash column chromatography, prep HPLC, or a combination of both.

(202) Method D.

(203) Amide coupling. To a solution of carboxylic acid (1 eq) in DCM (20 vol) under nitrogen were added oxalyl chloride (3 eq) and DMF (cat). The reaction mixture was stirred at ambient temperature for 30 minutes after which time the solvents were removed in vacuo. DCM (20 vol) or THF (20 vol) was added, followed by the required amine (1 to 3 eq) and triethylamine (2 eq) and the reaction mixture was stirred at ambient temperature. The reaction was monitored by LCMS to completion whereupon water was added. The reaction mixture was then extracted with DCM and the organic layer was washed with water, saturated aqueous NaCl, dried over Na2SO4 or MgSO4 and the solvent removed in vacuo to afford the crude product. Purification was carried out by flash column chromatography, prep HPLC, a combination of both or by trituration with an appropriate solvent.

(204) While some embodiments have been shown and described, various modifications and substitutions may be made thereto without departing from the spirit and scope of the invention. For example, for claim construction purposes, it is not intended that the claims set forth hereinafter be construed in any way narrower than the literal language thereof, and it is thus not intended that exemplary embodiments from the specification be read into the claims. Accordingly, it is to be understood that the present invention has been described by way of illustration and not limitations on the scope of the claims.