tetrabutylazanium;fluoride;hydrate

Conditions: temp:60.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A vial was charged with (S)-tert-butyl 3-bromo-7-iodo-5′H-spiro[chromeno[2,3-b]pyridine-5,4′-thiazole]-2′-ylcarbamate (0.194 g, 0.338 mmol), tetrakis(triphenylphosphine)palladium (0.039 g, 0.034 mmol), tetrabutylammonium fluoride hydrate (0.41 g, 1.689 mmol), and copper(I) iodide (6.43 mg, 0.034 mmol). Added THF (1.7 mL) followed by trimethyl((3-methyloxetan-3-yl)ethynyl)silane (0.10 mL, 0.51 mmol). The reaction was stirred at 60° C. for 30 min, then diluted with EtOAc and washed with water. The aqueous layer was extracted with EtOAc, and the combined organic layers were dried with sodium sulfate, filtered, and concentrated. The material was purified via column chromatography on silica gel (0-50% EtOAc:Hex) to afford (S)-tert-butyl 3-bromo-7-((3-methyloxetan-3-yl)ethynyl)-5′H-spiro[chromeno[2,3-b]pyridine-5,4′-thiazole]-2′-ylcarbamate (0.163 g, 0.30 mmol, 89% yield) as a yellow solid. washed with water The aqueous layer was extracted with EtOAc the combined organic layers were dried with sodium sulfate filtered concentrated The material was purified via column chromatography on silica gel (0-50% EtOAc:Hex)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 114 g (131 mmol) crude [[(3β)-25-[[(1,1-dimethylethyl)dimethylsilyl]oxy]cholesta-5,7,16-trien-23-yn-3-yl]oxy]dimethyl(1,1,2-trimethylpropyl)silane in 500 mL of THF was added 200 g (765 mmol) tetrabutylammonium fluoride hydrate with the aid of 500 mL of THF. After stirring at room temperature for 18 hr., the mixture was diluted with 500 mL of ethyl acetate and 500 mL of 10% aqueous ammonium chloride. The aqueous layer was extracted twice with 500 mL each of ethyl acetate. The combined organic layers were washed with 500 mL of 10% aqueous ammonium chloride, 500 mL of water, and 500 mL of brine. The organic layer was dried over MgSO4 and concentrated to a total weight of 525 g. The resulting suspension was stirred at room temperature for 2 hr and then stored in a freezer overnight. The solid was filtered, washed with cold ethyl acetate, and air-dried for 2 hr to give 27.8 g of crude (3β)-cholesta-5,7,16-trien-23-yne-3,25-diol. This was suspended in 270 mL of methanol and the mixture was refluxed for 10 min. Then, 55 mL waster was added dropwise over 10 min. The suspension was stirred at 60° C. for 1 h, and stored in a refrigerator overnight. The precipitate was filtered, washed with cold 100 mL 50% aqueous methanol, and dried in a vacuum oven at 45° C. to give 25.4 g (49.1% over four steps) of (3β)-cholesta-5,7,16-trien-23-yne-3,25-diol as white crystals: mp 185°-192° C. Anal. Calcd. for C27H38O2 : C, 82.18; H, 9.71. Found: C, 81.93; H, 9.74. The aqueous layer was extracted twice with 500 mL each of ethyl acetate The combined organic layers were washed with 500 mL of 10% aqueous ammonium chloride, 500 mL of water, and 500 mL of brine The organic layer was dried over MgSO4 concentrated to a total weight of 525 g The resulting suspension was stirred at room temperature for 2 hr stored in a freezer overnight The solid was filtered washed with cold ethyl acetate air-dried for 2 hr

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 114 g (131 mmol) crude [[(3β)-25-[[(1,1-dimethylethyl)dimethylsilyl]oxy]cholesta-5,7,16-trien-23-yn-3-yl]oxy]dimethyl(1,1,2-trimethylpropyl)silane in 500 mL of THF was added 200 g (765 mmol) tetrabutylammonium fluoride hydrate with the aid of 500 mL of THF. After stirring at room temperature for 18 hr., the mixture was diluted with 500 mL of ethyl acetate and 500 mL of 10% aqueous ammonium chloride. The aqueous layer was extracted twice with 500 mL each of ethyl acetate. The combined organic layers were washed with 500 mL of 10% aqueous ammonium chloride, 500 mL of water, and 500 mL of brine. The organic layer was dried over MgSO4 and concentrated to a total weight of 525 g. The resulting suspension was stirred at room temperature for 2 hr and then stored in a freezer overnight. The solid was filtered, washed with cold ethyl acetate, and air-dried for 2 hr to give 27.8 g of crude (3β)-cholesta-5,7,16-trien-23-yne-3,25-diol. This was suspended in 270 mL of methanol and the mixture was refluxed for 10 min. Then, 55 mL waster was added dropwise over 10 min. The suspension was stirred at 60° C. for 1 h, and stored in a refrigerator overnight. The precipitate was filtered, washed with cold 100 mL 50% aqueous methanol, and dried in a vacuum oven at 45° C. to give 25.4 g (49.1% over four steps) of (3β)-cholesta-5,7,16-trien-23-yne-3,25-diol as white crystals: mp 185°-192° C. Anal. Calcd. for C27H38O2 : C, 82.18; H, 9.71. Found: C, 81.93; H, 9.74. The aqueous layer was extracted twice with 500 mL each of ethyl acetate The combined organic layers were washed with 500 mL of 10% aqueous ammonium chloride, 500 mL of water, and 500 mL of brine The organic layer was dried over MgSO4 concentrated to a total weight of 525 g The resulting suspension was stirred at room temperature for 2 hr stored in a freezer overnight The solid was filtered washed with cold ethyl acetate air-dried for 2 hr

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: N,N-Di(1,1-dimethylethoxycarbonyl)-3-(1-(2,3-difluorophenyl)-1H-tetrazol-5-yl)-5-(2-(trimethylsilyl)ethynyl)pyridin-2-amine (713 mg, 1.2 mmol) was dissolved in methylene chloride (10 mL) and cooled to 0° C. To this was added tetrabutylammonium fluoride hydrate (163 mg, 625 μmol) in methylene chloride (1 mL). The reaction was loaded directly onto silica and chromatographed (10% ethyl acetate/hexane to 50% ethyl acetate/hexane) to give N,N-di(1,1-dimethylethoxycarbonyl)-3-(1-(2,3-difluorophenyl)-1H-tetrazol-5-yl)-5-ethynylpyridin-2-amine (496 mg, 1.0 mmol, 83%) as a yellow solid; 1H NMR (CDCl3): 1.37 (s, 18H), 3.35 (s, 1H), 7.30 (m, 1H), 7.40 (m, 2H), 7.89 (s, 1H), 8.75 (s, 1H). chromatographed (10% ethyl acetate/hexane to 50% ethyl acetate/hexane)

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: N,N-Di(1,1-dimethylethoxycarbonyl)-3-(1-(2,3-difluorophenyl)-1H-tetrazol-5-yl)-5-(2-(trimethylsilyl)ethynyl)pyridin-2-amine (713 mg, 1.2 mmol) was dissolved in methylene chloride (10 mL) and cooled to 0° C. To this was added tetrabutylammonium fluoride hydrate (163 mg, 625 μmol) in methylene chloride (1 mL). The reaction was loaded directly onto silica and chromatographed (10% ethyl acetate/hexane to 50% ethyl acetate/hexane) to give N,N-di(1,1-dimethylethoxycarbonyl)-3-(1-(2,3-difluorophenyl)-1H-tetrazol-5-yl)-5-ethynylpyridin-2-amine (496 mg, 1.0 mmol, 83%) as a yellow solid; 1H NMR (CDCl3): 1.37 (s, 18H), 3.35 (s, 1H), 7.30 (m, 1H), 7.40 (m, 2H), 7.89 (s, 1H), 8.75 (s, 1H). chromatographed (10% ethyl acetate/hexane to 50% ethyl acetate/hexane)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 2-bromobenzaldehyde (27.0 g, 146.5 mmol) and trimethylsilyl-trifluoromethane (25.0 g, 175.8 mmol) in tetrahydrofuran (300 ml) was added drop-wise tetrabutylammonium fluoride (1 M in tetrahydrofuran, 5 ml, 5 mmol) at 0° C. After the addition, the mixture was stirred at room temperature for 2 hours. Further tetrabutylammonium fluoride hydrate (49.0 g, 175.8 mmol) was added and the mixture was stirred for 30 minutes. The solvent was evaporated in vacuo. The residue was dissolved in dichloromethane (200 ml) and washed with hydrochloric acid (2N aqueous solution) (4×100 ml). The organic layer was separated, washed with Na2CO3 (10% aqueous solution) (50 ml), dried over anhydrous MgSO4 (s), filtered and evaporated in vacuo to afford the crude title compound as a yellow oil (45.0 g, ˜100%). After the addition the mixture was stirred for 30 minutes The solvent was evaporated in vacuo The residue was dissolved in dichloromethane (200 ml) washed with hydrochloric acid (2N aqueous solution) (4×100 ml) The organic layer was separated washed with Na2CO3 (10% aqueous solution) (50 ml) dried over anhydrous MgSO4 (s) filtered evaporated in vacuo

Conditions: temp:70.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 6-[[8-cyclopropyl-9-(2-trimethylsilylethoxymethyl)purin-6-yl]amino]-3-methyl-3-(trifluoromethyl)-2H-imidazo[1,5-a]pyridine-1,5-dione (8, 0.25 g, 0.47 mmol) in tetrahydrofuran (50 mL) was added tetrabutylammonium fluoride hydrate (0.61 g, 2.33 mmol). The mixture was heated at 70° C. for 4 h. After TLC showed consumption of 8, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The crude mixture was purified via column chromatography (silica, methanol/dichloromethane=2%) to afford 6-[(8-cyclopropyl-9H-purin-6-yl)amino]-3-methyl-3-(trifluoromethyl)-2H-imidazo[1,5-a]pyridine-1,5-dione (Cpd. No. 37) as yellow solid. Yield: 135 mg, 71%; MS (ESI) m/z 406.29 [M+1]+; 1H NMR (400 MHz, DMSO-d6) δ 13.24 (s, 1H), 10.55 (s, 1H), 8.80 (d, J=7.6 Hz, 1H), 8.64 (s, 1H), 8.49 (s, 1H), 7.10 (d, J=7.6 Hz, 1H), 2.16 (m, 1H), 2.15 (s, 3H), 1.12 (m, 4H). consumption of 8 the reaction mixture was diluted with water (50 mL) extracted with ethyl acetate (2×50 mL) The combined organic layers were dried over sodium sulfate concentrated under reduced pressure The crude mixture was purified via column chromatography (silica, methanol/dichloromethane=2%)

Conditions: temp:70.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 6-[[8-cyclopropyl-9-(2-trimethylsilylethoxymethyl)purin-6-yl]amino]-3-methyl-3-(trifluoromethyl)-2H-imidazo[1,5-a]pyridine-1,5-dione (8, 0.25 g, 0.47 mmol) in tetrahydrofuran (50 mL) was added tetrabutylammonium fluoride hydrate (0.61 g, 2.33 mmol). The mixture was heated at 70° C. for 4 h. After TLC showed consumption of 8, the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The crude mixture was purified via column chromatography (silica, methanol/dichloromethane=2%) to afford 6-[(8-cyclopropyl-9H-purin-6-yl)amino]-3-methyl-3-(trifluoromethyl)-2H-imidazo[1,5-a]pyridine-1,5-dione (Cpd. No. 37) as yellow solid. Yield: 135 mg, 71%; MS (ESI) m/z 406.29 [M+1]+; 1H NMR (400 MHz, DMSO-d6) δ 13.24 (s, 1H), 10.55 (s, 1H), 8.80 (d, J=7.6 Hz, 1H), 8.64 (s, 1H), 8.49 (s, 1H), 7.10 (d, J=7.6 Hz, 1H), 2.16 (m, 1H), 2.15 (s, 3H), 1.12 (m, 4H). consumption of 8 the reaction mixture was diluted with water (50 mL) extracted with ethyl acetate (2×50 mL) The combined organic layers were dried over sodium sulfate concentrated under reduced pressure The crude mixture was purified via column chromatography (silica, methanol/dichloromethane=2%)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a round bottomed flask was added N′-[4-(2,4-bis{[tert-butyl(dimethyl)silyl]oxy}phenyl)cyclohexylidene-4-methylbenzenesulfonohydrazide (100 mg), tetrahydrofuran (5 ml), glacial acetic acid (3 drops) and tetrabutylammonium fluoride hydrate (174 mg), and the resulting solution was stirred at room temperature for 2 hr. Saturated sodium hydrogen carbonate solution (10 ml) was added and stirring was continued for 1 hr. The reaction mixture was partitioned between water (10 ml) and ethyl acetate (20 ml), the layers were separated, and the aqueous phase was extracted with ethyl acetate (2×20 ml). The combined organic phases were washed with brine (10 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. Purification via flash chromatography (SiO2, ethyl acetate/petroleum ether, 7:3 v/v) furnished the title compound (62 mg, 100%) as a cream solid. δH ((CD3)2CO): 1.44-1.63 (2H, m), 1.92-2.01 (3H, m), 2.22-2.32 (1H, m), 2.35-2.43 (2H, m), 2.97 (3H, s), 3.09 (1H, tt), 6.28 (H, dd), 6.37 (1H, d), 6.87(1H, d), 7.38 (2H, d), 7.79 (2H, d), 7.96 (1H, s), 8.13 (1H, s), 8.99 (1H, s); m/z (ES+) 375 (M+H)+. stirring was continued for 1 hr The reaction mixture was partitioned between water (10 ml) and ethyl acetate (20 ml) the layers were separated the aqueous phase was extracted with ethyl acetate (2×20 ml) The combined organic phases were washed with brine (10 ml) dried over magnesium sulfate filtered concentrated in vacuo Purification via flash chromatography (SiO2, ethyl acetate/petroleum ether, 7:3 v/v)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a round bottomed flask was added N′-[4-(2,4-bis{[tert-butyl(dimethyl)silyl]oxy}phenyl)cyclohexylidene-4-methylbenzenesulfonohydrazide (100 mg), tetrahydrofuran (5 ml), glacial acetic acid (3 drops) and tetrabutylammonium fluoride hydrate (174 mg), and the resulting solution was stirred at room temperature for 2 hr. Saturated sodium hydrogen carbonate solution (10 ml) was added and stirring was continued for 1 hr. The reaction mixture was partitioned between water (10 ml) and ethyl acetate (20 ml), the layers were separated, and the aqueous phase was extracted with ethyl acetate (2×20 ml). The combined organic phases were washed with brine (10 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. Purification via flash chromatography (SiO2, ethyl acetate/petroleum ether, 7:3 v/v) furnished the title compound (62 mg, 100%) as a cream solid. δH ((CD3)2CO): 1.44-1.63 (2H, m), 1.92-2.01 (3H, m), 2.22-2.32 (1H, m), 2.35-2.43 (2H, m), 2.97 (3H, s), 3.09 (1H, tt), 6.28 (H, dd), 6.37 (1H, d), 6.87(1H, d), 7.38 (2H, d), 7.79 (2H, d), 7.96 (1H, s), 8.13 (1H, s), 8.99 (1H, s); m/z (ES+) 375 (M+H)+. stirring was continued for 1 hr The reaction mixture was partitioned between water (10 ml) and ethyl acetate (20 ml) the layers were separated the aqueous phase was extracted with ethyl acetate (2×20 ml) The combined organic phases were washed with brine (10 ml) dried over magnesium sulfate filtered concentrated in vacuo Purification via flash chromatography (SiO2, ethyl acetate/petroleum ether, 7:3 v/v)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a round bottomed flask equipped with magnetic stirrer was added benzyl [4-(2,4-bis{[tert-butyl(dimethyl)silyl]oxy}phenyl)cyclohexylidene]acetate (232 mg), tetrahydrofuran (5 ml), tetrabutylammonium fluoride hydrate (429 mg) and glacial acetic acid (94 μl). After stirring at room temperature for 0.5 hr, the solvent was removed in vacuo and the residue partitioned between ethyl acetate (100 ml) and water (20 ml). The layers were separated and the aqueous phase was extracted with ethyl acetate (50 ml). The combined organic phases were washed with water (50 ml), brine (20 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. Purification via flash chromatography (SiO2, ethyl acetate/petroleum ether, 1:3 v/v) furnished the title compound (100 mg, 72%) as a cream solid. δH (CD3OD): 1.59-1.67 (2H, m), 1.98-2.13 (4H, m), 2.37-2.48 (2H, m), 3.14 (1H, ft), 5.16 (2H, s), 5.75 (1H, s), 6.26 (1H, dd), 6.30 (1H, d), 6.89 (1H, d), 7.30-7.44 (5H, m); m/z (ES+) 337 (M−H)−. To a round bottomed flask equipped with magnetic stirrer the solvent was removed in vacuo the residue partitioned between ethyl acetate (100 ml) and water (20 ml) The layers were separated the aqueous phase was extracted with ethyl acetate (50 ml) The combined organic phases were washed with water (50 ml), brine (20 ml) dried over magnesium sulfate filtered concentrated in vacuo Purification via flash chromatography (SiO2, ethyl acetate/petroleum ether, 1:3 v/v)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: The compound (S)-4-[3-(tert-Butyldimethylsilanyloxy)-2-methylpropyl]-6-methoxy-4H-benzo[1,4]oxazin-3-one (111MF32) (4.03 g, 12.0 mmol) and tetrabutylammonium flouride monohydrate (4.46 g, 16.0 mmol) were mixed according to GP8. CC (SiO2; Heptane/EtOAc 4:1-4) gave the title compound (111MF34) (2.70 g, 100%). 1H NMR (CDCl3) δ 6.91 (d, J=8.8 Hz, 1H), 6.63 (d, J=2.8 Hz, 1H), 6.54 (dd, J=2.8 Hz, J=8.8 Hz, 1H), 4.57 (s, 2H), 4.19 (dd, J=9.4 Hz, J=14.6 Hz, 1H), 3.78 (s, 3H), 3.56-3.39 (m, 3H), 2.84 (s, 1H), 2.09-1.99 (m, 1H), 1.05 (d, J=7.2 Hz, 3H); 13C NMR (CDCl3) 165.8, 155.5, 139.5, 129.4, 117.4, 107.7, 103.0, 67.7, 63.8, 56.0, 43.6, 34.1, 15.0.