N-diazo-1,1,1-trifluoromethanesulfonamide

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

Procedure: 5'-O-(t-Butyldimethylsilyl)-2',3'-dideoxy-3'-(N,N-dimethylamino-methylene)-2'-oxo-uridine (1.78 g, 4.5 mmol), from Step 6, was added to the trifluoromethanesulfonyl azide solution and the reaction mixture was heated at 60° C. for 2.5 h under a nitrogen atmosphere. The solvents were evaporated in vacuo to a syrup (2.8 g) which was dissolved in 10 mL of methylene chloride and purified by flash chromatography (5 psi) on a silica gel column (2×35 cm) eluted with 200 mL of methylene chloride, 200 mL of 1% methanol in methylene chloride, 200 mL of 2% methanol in methylene chloride and 3% methanol in methylene chloride to give 1.27 g (77% yield) of the title compound. MS DCI: 367 (M+H)+. The solvents were evaporated in vacuo to a syrup (2.8 g) which was dissolved in 10 mL of methylene chloride purified by flash chromatography (5 psi) on a silica gel column (2×35 cm) eluted with 200 mL of methylene chloride, 200 mL of 1% methanol in methylene chloride, 200 mL of 2% methanol in methylene chloride

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

Procedure: At 0° C., 1.19 g(18.3 mmol) of sodium azide in a mixture of 3 ml of water and 5 ml of dichloromethane are mixed dropwise with 1.046 g (mmol) of trifluoromethanesulphonic anhydride, and the resulting mixture is stirred at 0° C. for 2 hours. The phases are separated, the aqueous phase is extracted twice with dichloromethane and the combined organic phases are washed with saturated sodium bicarbonate solution. 386 mg (2 mmol) of 3-amino-6-phenyl-hexan-2-ol are dissolved in 2 ml of methanol and admixed successively with 10 mg of copper sulphate and 10 mg of potassium carbonate. The solution of the triflyl azide in dichloromethane is added and the mixture is made up to a total volume of 20 ml using dichloromethane. The mixture is stirred at room temperature for 15 hours, extracted with dilute ammonia solution and with EDTA solution, dried over sodium sulphate and concentrated using a rotary evaporator. This gives 331 mg of an oil which is dissolved in 1 ml of DMSO and admixed with a solution of 75 mg (3.5 mmol) of sodium in 1 ml of ethanol. 197 mg (2.34 mmol) of cyanoacetamide and, after 30 minutes, 2 ml of ethanol are added, and the mixture is stirred at room temperature for 15 hours. The mixture is poured into 30 ml of ice-water and, after addition of 3 ml of 1N HCl, extracted three times with dichloromethane and once with ethyl acetate. Chromatographic purification gives 72 mg (12%) of a foam. The phases are separated the aqueous phase is extracted twice with dichloromethane the combined organic phases are washed with saturated sodium bicarbonate solution The mixture is stirred at room temperature for 15 hours extracted with dilute ammonia solution and with EDTA solution dried over sodium sulphate concentrated a rotary evaporator This gives 331 mg of an oil which is dissolved in 1 ml of DMSO the mixture is stirred at room temperature for 15 hours extracted three times with dichloromethane Chromatographic purification

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

Procedure: At 0° C., 1.19 g(18.3 mmol) of sodium azide in a mixture of 3 ml of water and 5 ml of dichloromethane are mixed dropwise with 1.046 g (mmol) of trifluoromethanesulphonic anhydride, and the resulting mixture is stirred at 0° C. for 2 hours. The phases are separated, the aqueous phase is extracted twice with dichloromethane and the combined organic phases are washed with saturated sodium bicarbonate solution. 386 mg (2 mmol) of 3-amino-6-phenyl-hexan-2-ol are dissolved in 2 ml of methanol and admixed successively with 10 mg of copper sulphate and 10 mg of potassium carbonate. The solution of the triflyl azide in dichloromethane is added and the mixture is made up to a total volume of 20 ml using dichloromethane. The mixture is stirred at room temperature for 15 hours, extracted with dilute ammonia solution and with EDTA solution, dried over sodium sulphate and concentrated using a rotary evaporator. This gives 331 mg of an oil which is dissolved in 1 ml of DMSO and admixed with a solution of 75 mg (3.5 mmol) of sodium in 1 ml of ethanol. 197 mg (2.34 mmol) of cyanoacetamide and, after 30 minutes, 2 ml of ethanol are added, and the mixture is stirred at room temperature for 15 hours. The mixture is poured into 30 ml of ice-water and, after addition of 3 ml of 1N HCl, extracted three times with dichloromethane and once with ethyl acetate. Chromatographic purification gives 72 mg (12%) of a foam. The phases are separated the aqueous phase is extracted twice with dichloromethane the combined organic phases are washed with saturated sodium bicarbonate solution The mixture is stirred at room temperature for 15 hours extracted with dilute ammonia solution and with EDTA solution dried over sodium sulphate concentrated a rotary evaporator This gives 331 mg of an oil which is dissolved in 1 ml of DMSO the mixture is stirred at room temperature for 15 hours extracted three times with dichloromethane Chromatographic purification

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

Procedure: Boc-Lys(N3)OH was synthesized from Boc-LysOH by using procedures for Cu(II)-catalyzed diazo-transfer to amines. (Alper, P. B.; Hung, S.-C.; Wong, C.-H. Tetrahedron Lett. 1996, 37, 6029-6032.) Boc-LysOH (4.38 g, 17.8 mmol), CuSO4.5H2O (44.7 mg, 178.2 □mol) and K2CO3 (3.69 g, 26.8 mmol) were dissolved in H2O (57 mL) and MeOH (114 mL). N3Tf (triflyl azide, 17.8 mmol in 150 mL CH2Cl2) was added, and the reaction mixture was stirred overnight. MeOH was removed under reduced pressure. The resulting aqueous solution (50 mL) was diluted with H2O (0.10 L), and the pH was adjusted to 6.0 with HCl. Sodium phosphate buffer (150 mL, 0.25 M, pH 6.2) was added, and the solution was washed with EtOAc (3×) to remove the triflic amine byproduct. The pH of the washed aqueous layer was adjusted to 2.0 with HCl, and the resulting solution was washed with EtOAc (4×) to extract the desired product. The organic layer was then dried over MgSO4(s) and filtered, and the solvent was removed under reduced pressure to yield Boc-Lys(N3)OH as a clear oil (50%). Spectral data. 1H NMR (500 MHz, CDCl3) δ:10.93 (broad s, 1H), 5.11 (d, J=8.3 Hz, 1H), 4.34 (m, 1H), 3.30 (d, J=7.0 Hz, 2H), 1.80 (m, 2H), 1.65 (m, 2H), 1.49 (m, 2H), 1.46 (s, 9H) ppm; 13C NMR (125 MHz, CDCl3) δ:179.96, 156.30, 97.44, 55.79, 53.79, 34.66, 31.07, 30.96, 25.20 ppm; MS (ESI) m/z 295.1383 (MNa+[C11H20N4O4Na]=295.1382). MeOH was removed under reduced pressure The resulting aqueous solution (50 mL) was diluted with H2O (0.10 L) Sodium phosphate buffer (150 mL, 0.25 M, pH 6.2) was added the solution was washed with EtOAc (3×) to remove the triflic amine byproduct the resulting solution was washed with EtOAc (4×) to extract the desired product The organic layer was then dried over MgSO4(s) filtered the solvent was removed under reduced pressure

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

Procedure: Boc-Lys(N3)OH was synthesized from Boc-LysOH by using procedures for Cu(II)-catalyzed diazo-transfer to amines. (Alper, P. B.; Hung, S.-C.; Wong, C.-H. Tetrahedron Lett. 1996, 37, 6029-6032.) Boc-LysOH (4.38 g, 17.8 mmol), CuSO4.5H2O (44.7 mg, 178.2 □mol) and K2CO3 (3.69 g, 26.8 mmol) were dissolved in H2O (57 mL) and MeOH (114 mL). N3Tf (triflyl azide, 17.8 mmol in 150 mL CH2Cl2) was added, and the reaction mixture was stirred overnight. MeOH was removed under reduced pressure. The resulting aqueous solution (50 mL) was diluted with H2O (0.10 L), and the pH was adjusted to 6.0 with HCl. Sodium phosphate buffer (150 mL, 0.25 M, pH 6.2) was added, and the solution was washed with EtOAc (3×) to remove the triflic amine byproduct. The pH of the washed aqueous layer was adjusted to 2.0 with HCl, and the resulting solution was washed with EtOAc (4×) to extract the desired product. The organic layer was then dried over MgSO4(s) and filtered, and the solvent was removed under reduced pressure to yield Boc-Lys(N3)OH as a clear oil (50%). Spectral data. 1H NMR (500 MHz, CDCl3) δ:10.93 (broad s, 1H), 5.11 (d, J=8.3 Hz, 1H), 4.34 (m, 1H), 3.30 (d, J=7.0 Hz, 2H), 1.80 (m, 2H), 1.65 (m, 2H), 1.49 (m, 2H), 1.46 (s, 9H) ppm; 13C NMR (125 MHz, CDCl3) δ:179.96, 156.30, 97.44, 55.79, 53.79, 34.66, 31.07, 30.96, 25.20 ppm; MS (ESI) m/z 295.1383 (MNa+[C11H20N4O4Na]=295.1382). MeOH was removed under reduced pressure The resulting aqueous solution (50 mL) was diluted with H2O (0.10 L) Sodium phosphate buffer (150 mL, 0.25 M, pH 6.2) was added the solution was washed with EtOAc (3×) to remove the triflic amine byproduct the resulting solution was washed with EtOAc (4×) to extract the desired product The organic layer was then dried over MgSO4(s) filtered the solvent was removed under reduced pressure

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

Procedure: To an ice-cold mixture of methyl 3-amino-2-hydroxypropionate hydrochloride (19) (27.1 g, 0.17 mol) in water (100 mL) and dichloromethane (100 mL) was added sodium carbonate (19.82 g, 0.187 mol) and CuSO4. 5H2O (0.3 g, 1.2 mmol) with stirring. Triflylazide from experiment 10(B)(i) (crude, 44.6 g, 0.25 mol) was added dropwise keeping the reaction temperature at 0-5° C. Methanol (about 100 mL) was added until the reaction mixture became homogeneous. The reaction mixture was allowed to come to room temperature and then stirred for 20 h. Water (500 mL) was added and the aqueous solution thoroughly extracted with dichloromethane (5×100 mL). The combined organic layers were washed with water (2×200 mL), saturated sodium carbonate (2×200 mL), and then dried (anhydrous sodium sulfate). After removal of the solvent, the residue was chromatographed over silica gel. Elution with 30% ethyl acetate in hexanes furnished the product as a colorless oil (14.0 g; Yield: 57%) Rf 0.57 (30% ethyl acetate in hexanes). at 0-5° C to come to room temperature stirred for 20 h the aqueous solution thoroughly extracted with dichloromethane (5×100 mL) The combined organic layers were washed with water (2×200 mL), saturated sodium carbonate (2×200 mL) dried (anhydrous sodium sulfate) After removal of the solvent the residue was chromatographed over silica gel Elution with 30% ethyl acetate in hexanes

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

Procedure: To an ice-cold mixture of methyl 3-amino-2-hydroxypropionate hydrochloride (19) (27.1 g, 0.17 mol) in water (100 mL) and dichloromethane (100 mL) was added sodium carbonate (19.82 g, 0.187 mol) and CuSO4. 5H2O (0.3 g, 1.2 mmol) with stirring. Triflylazide from experiment 10(B)(i) (crude, 44.6 g, 0.25 mol) was added dropwise keeping the reaction temperature at 0-5° C. Methanol (about 100 mL) was added until the reaction mixture became homogeneous. The reaction mixture was allowed to come to room temperature and then stirred for 20 h. Water (500 mL) was added and the aqueous solution thoroughly extracted with dichloromethane (5×100 mL). The combined organic layers were washed with water (2×200 mL), saturated sodium carbonate (2×200 mL), and then dried (anhydrous sodium sulfate). After removal of the solvent, the residue was chromatographed over silica gel. Elution with 30% ethyl acetate in hexanes furnished the product as a colorless oil (14.0 g; Yield: 57%) Rf 0.57 (30% ethyl acetate in hexanes). at 0-5° C to come to room temperature stirred for 20 h the aqueous solution thoroughly extracted with dichloromethane (5×100 mL) The combined organic layers were washed with water (2×200 mL), saturated sodium carbonate (2×200 mL) dried (anhydrous sodium sulfate) After removal of the solvent the residue was chromatographed over silica gel Elution with 30% ethyl acetate in hexanes

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

Procedure: To a solution of tert-butyl (2R,5R)-5-(aminomethyl)-2-[2-(2-{[N-(methoxycarbonyl)-β-phenyl-L-phenylalanyl]amino}phenyl)ethyl]morpholine-4-carboxylate (1 eq.) in CH2Cl2 (0.4M) was added copper(II) sulfate (0.02 eq.) and sodium bicarbonate (1 eq.) as a solution in water (NaHCO3/CuSO4)) (1M). To that mixture was added triflic azide (3.65 eq.) as a solution in CH2Cl2 (0.4M). The heterogenous mixture was homogenized by the addition of methanol and the reaction was stirred at room temperature for 30 minutes. Upon completion by LCMS, solvents were evaporated and the evaporation residue was used as such for next step. Upon completion by LCMS, solvents were evaporated

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

Procedure: Following the method described by Aubé and coworkers, supra, a solution of NaN3 (0.88 g, 13.7 mmol) in H2O (2.5 mL) was cooled to 0° C. and CH2Cl2 (4 mL) was added. Whilst the mixture was stirring vigorously, Tf2O (0.47 mL, 2.26 mmol) was added dropwise. The resulting solution was stirred for an additional 2 h. The mixture was then placed in a separating funnel, and the organic layer was removed. The aqueous phase was extracted further with CH2Cl2 (2×3 mL). The organic layers were combined, washed with saturated Na2CO3, dried over Na2SO4(s), and filtered. The resulting solution of TfN3 was added dropwise to solution of methyl 2-amino-6-{[(benzyloxy)carbonyl]amino}hexanoate hydrochloride salt (0.46 g, 1.40 mmol) and DMAP (0.75 g, 6.16 mmol) in CH2Cl2 (5 mL). The resulting solution was stirred overnight under Ar(g). The mixture was concentrated under reduced pressure and purified by silica gel flash chromatography, eluting with 20% EtOAc/hexanes, to give azide 2f as an oil (0.43 g, 1.34 mmol, 96% yield). The resulting solution was stirred for an additional 2 h The mixture was then placed in a separating funnel the organic layer was removed The aqueous phase was extracted further with CH2Cl2 (2×3 mL) washed with saturated Na2CO3 dried over Na2SO4(s) filtered The mixture was concentrated under reduced pressure purified by silica gel flash chromatography eluting with 20% EtOAc/hexanes

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

Procedure: Following a method described by Aubé and coworkers [S. K. Ramanathan, J. Keeler, H.-L. Lee, D. S. Reddy, G. Lushington, J. Aubé, Org. Lett. 2005, 7, 1059-1062], a solution of NaN3 (0.88 g, 13.7 mmol) in H2O (2.5 mL) was cooled to 0° C. and CH2Cl2 (4 mL) was added. Whilst the mixture was stirring vigorously, Tf2O (CF3SO2OSO2CF3, 0.47 mL, 2.26 mmol) was added dropwise. The resulting solution was stirred for an additional 2 h. The mixture was then placed in a separating funnel, and the organic layer was removed. The aqueous phase was extracted further with CH2Cl2 (2×3 mL). The organic layers were combined, washed with saturated Na2CO3, dried over Na2SO4(s), and filtered. The resulting solution of TfN3 was added dropwise to a solution of methyl 2-amino-3-(benzylsulfanyl)propanoate hydrochloride salt (0.37 g, 1.40 mmol) and DMAP (0.75 g, 6.16 mmol) in CH2Cl2 (5 mL). The resulting solution was stirred overnight under Ar(g). The mixture was concentrated under reduced pressure and purified by silica gel flash chromatography, eluting with CH2Cl2, to give azide 2e as an oil (0.26 g, 1.05 mmol, 75% yield). The resulting solution was stirred for an additional 2 h The mixture was then placed in a separating funnel the organic layer was removed The aqueous phase was extracted further with CH2Cl2 (2×3 mL) washed with saturated Na2CO3 dried over Na2SO4(s) filtered The mixture was concentrated under reduced pressure purified by silica gel flash chromatography eluting with CH2Cl2

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

Procedure: Following a method described by Aubé and coworkers [S. K. Ramanathan, J. Keeler, H.-L. Lee, D. S. Reddy, G. Lushington, J. Aubé, Org. Lett. 2005, 7, 1059-1062], a solution of NaN3 (0.88 g, 13.7 mmol) in H2O (2.5 mL) was cooled to 0° C. and CH2Cl2 (4 mL) was added. Whilst the mixture was stirring vigorously, Tf2O (CF3SO2OSO2CF3, 0.47 mL, 2.26 mmol) was added dropwise. The resulting solution was stirred for an additional 2 h. The mixture was then placed in a separating funnel, and the organic layer was removed. The aqueous phase was extracted further with CH2Cl2 (2×3 mL). The organic layers were combined, washed with saturated Na2CO3, dried over Na2SO4(s), and filtered. The resulting solution of TfN3 was added dropwise to a solution of methyl 2-amino-3-(benzylsulfanyl)propanoate hydrochloride salt (0.37 g, 1.40 mmol) and DMAP (0.75 g, 6.16 mmol) in CH2Cl2 (5 mL). The resulting solution was stirred overnight under Ar(g). The mixture was concentrated under reduced pressure and purified by silica gel flash chromatography, eluting with CH2Cl2, to give azide 2e as an oil (0.26 g, 1.05 mmol, 75% yield). The resulting solution was stirred for an additional 2 h The mixture was then placed in a separating funnel the organic layer was removed The aqueous phase was extracted further with CH2Cl2 (2×3 mL) washed with saturated Na2CO3 dried over Na2SO4(s) filtered The mixture was concentrated under reduced pressure purified by silica gel flash chromatography eluting with CH2Cl2

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

Procedure: 5'-O-(t-Butyldimethylsilyl)-2',3'-dideoxy-3'-(N,N-dimethylamino-methylene)-2'-oxo-uridine (1.78 g, 4.5 mmol), from Step 6, was added to the trifluoromethanesulfonyl azide solution and the reaction mixture was heated at 60° C. for 2.5 h under a nitrogen atmosphere. The solvents were evaporated in vacuo to a syrup (2.8 g) which was dissolved in 10 mL of methylene chloride and purified by flash chromatography (5 psi) on a silica gel column (2×35 cm) eluted with 200 mL of methylene chloride, 200 mL of 1% methanol in methylene chloride, 200 mL of 2% methanol in methylene chloride and 3% methanol in methylene chloride to give 1.27 g (77% yield) of the title compound. MS DCI: 367 (M+H)+. The solvents were evaporated in vacuo to a syrup (2.8 g) which was dissolved in 10 mL of methylene chloride purified by flash chromatography (5 psi) on a silica gel column (2×35 cm) eluted with 200 mL of methylene chloride, 200 mL of 1% methanol in methylene chloride, 200 mL of 2% methanol in methylene chloride