4-hydroxybutanenitrile

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added 1M sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added IM sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added 1M sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added 1M sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes

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

Procedure: To a solution of 3-cyano-1-n-propanol (16.85 g, 198 mmol, prepared by method of Blicke, et al., J. Org. Chem. 1961, 26, 3685) in dichloromethane (250 mL) were added triethylamine (80.1 g, 792 mmol) and trityl chloride (82.8 g, 297 mmol) at room temperature and the mixture was stirred for 16 h. To the reaction mixture was added methanol (25 mL) and stirred for 1 h. After concentration, the residue was purified on a column of silica gel using ethyl acetate and hexanes as eluent to give 40 g (61.7%) of the desired product. stirred for 1 h After concentration the residue was purified on a column of silica gel

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

Procedure: To a solution of 3-cyano-1-n-propanol (16.85 g, 198 mmol, prepared by method of Blicke, et al., J. Org. Chem. 1961, 26, 3685) in dichloromethane (250 mL) were added triethylamine (80.1 g, 792 mmol) and trityl chloride (82.8 g, 297 mmol) at room temperature and the mixture was stirred for 16 h. To the reaction mixture was added methanol (25 mL) and stirred for 1 h. After concentration, the residue was purified on a column of silica gel using ethyl acetate and hexanes as eluent to give 40 g (61.7%) of the desired product. stirred for 1 h After concentration the residue was purified on a column of silica gel

Conditions: temp:100.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of cholesteryl tosylate (2.8 g, 5.1 mmol) and 4-hydroxybutanenitrile (0.61 g, 7.1 mmol) in toluene was stirred at 120° C. for 24 h and then the solvent was evaporated in vacuo. The obtained crude 4-(cholest-5-en-3β-yloxy)butanenitrile was dissolved in isopropyl alcohol (30 ml), aqueous NaOH (12%; 100 ml) was added and the mixture was stirred at 100° C. for 3 days. The mixture was cooled to room temperature, aqueous hydrochloric acid (10%, 100 ml) was added and the product was taken in diethyl ether (4×30 ml). The combined extracts were dried over anhydrous MgSO4 and concentrated in vacuo. Flash chromatography of the residue on silica gel column (320 ml) in toluene-ethyl acetate (gradient: 0-40% ethyl acetate, 10 ml/min, 160 min) afforded 4-(Cholest-5-en-3β-yloxy)butanoic acid (general formula IX, where n2=3; 345 mg, 10%). 1H NMR (400 MHz, CHCl3) δ (ppm): 5.32-5.39 m, 1H (H-6); 3.55 td, J=5.90, 1.40 Hz, 2H (CH2-1′); 3.17 tt, J=11.21, 4.39 Hz, 1H (H-3); 2.49 t, J=7.01 Hz, 2H (CH2-3′); 2.36 ddd, J=13.17, 4.71, 2.08 Hz, 1H (H-4a); 2.14-2.24 m, 1H (H-4b); 1.93-2.05 m, 2H (CH2-1, H-12); 1.90 t, J=6.20 Hz, 2H (CH2-2′); 1.78-1.88 m, 3H (H-2, H-7, H-16); 1.02-1.63 m, 22H (cholesterol); 1.01 s, 3H (CH3-19); 0.92 d, J=6.57 Hz, 3H (CH3-21); 0.88 d, J=1.77 Hz, 3H (CH3-26); 0.86 d, J=1.77 Hz, 3H (CH3-27); 0.68 s, 3H (CH3-18); 13C NMR (101 MHz, CHCl3) δ (ppm): 178.28 (C-1′) 140.76 (C-5) 121.69 (C-6) 79.31 (C-3) 66.88 (C-4′) 56.77 (C-14) 56.16 (C-17) 50.18 (C-9) 42.31 (C-13) 39.78 (C-12) 39.51 (C-24) 38.98 (C-4) 37.18 (C−1) 36.86 (C-10) 36.18 (C-22) 35.78 (C-20) 31.94 (C-7) 31.88 (C-8) 31.37 (C-2′) 28.29 (C-2) 28.23 (C-16) 28.01 (C-25) 25.06 (C-3′) 24.28 (C-15) 23.83 (C-23) 22.81 (C-26) 22.55 (C-27) 21.06 (C-11) 19.36 (C-19) 18.71 (C-21) 11.86 (C-18); for C31H52O3 monoisotopic mass: calculated: 472.4. found: MS ESI m/z: 495.4 [M+Na]+. for C31H52O3 calculated: 78.76% C; 11.09% H. found: 78.80% C; 11.18% H; IR (CHCl3): 3517, 2718, 1741, 1710, 1415, 1285 (COOH); 1134, 1105, 1070, 1083, 841 (ether); 1670, 1334 (C═C); 2954, 2869, 1445, 1366 (Me); [α]D-27.2° (c 0.136; CHCl3). the solvent was evaporated in vacuo The obtained crude 4-(cholest-5-en-3β-yloxy)butanenitrile The mixture was cooled to room temperature The combined extracts were dried over anhydrous MgSO4 concentrated in vacuo

Conditions: temp:100.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of cholesteryl tosylate (2.8 g, 5.1 mmol) and 4-hydroxybutanenitrile (0.61 g, 7.1 mmol) in toluene was stirred at 120° C. for 24 h and then the solvent was evaporated in vacuo. The obtained crude 4-(cholest-5-en-3β-yloxy)butanenitrile was dissolved in isopropyl alcohol (30 ml), aqueous NaOH (12%; 100 ml) was added and the mixture was stirred at 100° C. for 3 days. The mixture was cooled to room temperature, aqueous hydrochloric acid (10%, 100 ml) was added and the product was taken in diethyl ether (4×30 ml). The combined extracts were dried over anhydrous MgSO4 and concentrated in vacuo. Flash chromatography of the residue on silica gel column (320 ml) in toluene-ethyl acetate (gradient: 0-40% ethyl acetate, 10 ml/min, 160 min) afforded 4-(Cholest-5-en-3β-yloxy)butanoic acid (general formula IX, where n2=3; 345 mg, 10%). 1H NMR (400 MHz, CHCl3) δ (ppm): 5.32-5.39 m, 1H (H-6); 3.55 td, J=5.90, 1.40 Hz, 2H (CH2-1′); 3.17 tt, J=11.21, 4.39 Hz, 1H (H-3); 2.49 t, J=7.01 Hz, 2H (CH2-3′); 2.36 ddd, J=13.17, 4.71, 2.08 Hz, 1H (H-4a); 2.14-2.24 m, 1H (H-4b); 1.93-2.05 m, 2H (CH2-1, H-12); 1.90 t, J=6.20 Hz, 2H (CH2-2′); 1.78-1.88 m, 3H (H-2, H-7, H-16); 1.02-1.63 m, 22H (cholesterol); 1.01 s, 3H (CH3-19); 0.92 d, J=6.57 Hz, 3H (CH3-21); 0.88 d, J=1.77 Hz, 3H (CH3-26); 0.86 d, J=1.77 Hz, 3H (CH3-27); 0.68 s, 3H (CH3-18); 13C NMR (101 MHz, CHCl3) δ (ppm): 178.28 (C-1′) 140.76 (C-5) 121.69 (C-6) 79.31 (C-3) 66.88 (C-4′) 56.77 (C-14) 56.16 (C-17) 50.18 (C-9) 42.31 (C-13) 39.78 (C-12) 39.51 (C-24) 38.98 (C-4) 37.18 (C−1) 36.86 (C-10) 36.18 (C-22) 35.78 (C-20) 31.94 (C-7) 31.88 (C-8) 31.37 (C-2′) 28.29 (C-2) 28.23 (C-16) 28.01 (C-25) 25.06 (C-3′) 24.28 (C-15) 23.83 (C-23) 22.81 (C-26) 22.55 (C-27) 21.06 (C-11) 19.36 (C-19) 18.71 (C-21) 11.86 (C-18); for C31H52O3 monoisotopic mass: calculated: 472.4. found: MS ESI m/z: 495.4 [M+Na]+. for C31H52O3 calculated: 78.76% C; 11.09% H. found: 78.80% C; 11.18% H; IR (CHCl3): 3517, 2718, 1741, 1710, 1415, 1285 (COOH); 1134, 1105, 1070, 1083, 841 (ether); 1670, 1334 (C═C); 2954, 2869, 1445, 1366 (Me); [α]D-27.2° (c 0.136; CHCl3). the solvent was evaporated in vacuo The obtained crude 4-(cholest-5-en-3β-yloxy)butanenitrile The mixture was cooled to room temperature The combined extracts were dried over anhydrous MgSO4 concentrated in vacuo

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

Procedure: A mixture of 4-hydroxybutyronitrile (24.2 mg, 285 μmol) [Organometallics (1996), 15(4), 1236-41], KOtBu (34.8 mg, 311 μmol), and DME was stirred at rt, followed by the addition of 4-(7-Fluoro-quinazolin-4-yl)-piperidine-1-carboxylic acid tert-butyl ester (48.8 mg, 147 μmol) (prepared as described in Example 65b). The resulting homogeneous solution was stirred at rt for 2 hr, and was then directly loaded onto a 5 g Jones silica cartridge pre-equilibrated with 9:1 DCM/acetone, and eluted with 9:1→8:2 DCM/acetone to afford the title intermediate (24.5 mg, 42%) as a colorless oil. LC/MS (ESI) calcd mass 396.2, found 397.1 (MH)+. The resulting homogeneous solution was stirred at rt for 2 hr eluted with 9:1→8:2 DCM/acetone

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added 1M sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added 1M sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes

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

Procedure: To 2,5-dibromopyridine (500 mg, 2.1 mmol) and 3-cyano-1-propanol (270 mg, 3.1 mmol) in DMSO (2 mL) was added 1M sodium hexamethyldisilazide (2.1 mL, 2.1 mmol). The reaction was stirred at room temperature overnight. EtOAc was added to the reaction and the mixture was washed with water (2×10 mL). The EtOAc fraction was dried over anhydrous sodium sulfate and evaporated on a rotary evaporator. The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes to give 200 mg of 5-bromo-2-cyanopropyloxypyridine as a pale yellow solid (39.3%). the mixture was washed with water (2×10 mL) The EtOAc fraction was dried over anhydrous sodium sulfate evaporated on a rotary evaporator The crude product was purified by flash column chromatography over silica gel using 40%EtOAc/hexanes