35909-92-3

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

Procedure: N-Benzyloxycarbonyl phenylalanine methyl ester is reduced with DIBAL to the aldehyde using the procedure of Boger et al., J. Med. Chem. (1985) 28, 1779, which is homologated with the sodio derivative of trimethyl phosphonoacetate using the procedure of Wadsworth and Emmons, Org. Synth. (1965) 45, 44 to give methyl 4-(benzyloxycarbonyl)amino-5-phenyl-2-pentenoate.

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

Procedure: 20.0 g (92.73 mmol) of (L)-phenylalanine methyl ester hydrochloride was suspended in 93 ml of toluene, and 15.82 g (92.73 mmols) of benzyl chloroformate was added thereto. 130 ml of an aqueous solution of 1 M sodium carbonate was dropwise added thereto, while keeping it at 7° C. or lower, and stirred for 3 hours. After this was subjected to phase separation, the resulting organic layer was washed with 60 ml of 0.1 N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate, and then dried with anhydrous sodium sulfate. This was filtered and concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound. The yield was 95.8%. at 7° C. After this was subjected to phase separation the resulting organic layer was washed with 60 ml of 0.1 N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate dried with anhydrous sodium sulfate This was filtered concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound

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

Procedure: 20.0 g (92.73 mmol) of (L)-phenylalanine methyl ester hydrochloride was suspended in 93 ml of toluene, and 15.82 g (92.73 mmols) of benzyl chloroformate was added thereto. 130 ml of an aqueous solution of 1 M sodium carbonate was dropwise added thereto, while keeping it at 7° C. or lower, and stirred for 3 hours. After this was subjected to phase separation, the resulting organic layer was washed with 60 ml of 0.1 N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate, and then dried with anhydrous sodium sulfate. This was filtered and concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound. The yield was 95.8%. at 7° C. After this was subjected to phase separation the resulting organic layer was washed with 60 ml of 0.1 N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate dried with anhydrous sodium sulfate This was filtered concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound

Conditions: temp:4.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: After 1.80 g of Z-Phe was dissolved in 20 mL of MeOH, 73 mg of DMAP, 1.38 g of WSCD HCl were added at 0° C., followed by stirring at 4° C. for 12 hours. The solvent was concentrated and the concentrate was dissolved in AcOEt, followed by washing with 1N HCl aq., satd. NaHCO3 aq., and satd. NaCl aq. After drying over Na2SO4, the solvent was concentrated to give Z-Phe-OMe as oil. After dissolving in 20 mL of dry THF, 196 mg of LiBH4 was added thereto and the mixture was stirred at room temperature for 15 hours. The solvent was concentrated and the concentrate was dissolved in AcOEt and the residue was washed with 1N HCl aq., satd. NaHCO3 aq., and satd. NaCl aq. After drying over Na2SO4, the solvent was concentrated and ether-petroleum ether was added to give 1.45 g (yield 85%) of Z-Phe-ol as precipitates. After 60 mg of 60% NaH was suspended in 10 mL of dry THF, 285 mg of Z-Phe-ol, 264 mg of 18-crown-6 and 1.48 mL of tert-butyl bromoacetate were added at 0° C. While returning to room temperature, the mixture was stirred for 15 hours. The solvent was concentrated the concentrate was dissolved in AcOEt by washing with 1N HCl aq. NaCl aq. After drying over Na2SO4 the solvent was concentrated

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

Procedure: To a tetrahydrofuran solution (1 ml) of the compound prepared in Example 6 (174 mg, 0.5 mmol), were succcessively added dropwise at 0° C. in a nitrogen atmosphere benzyl bromide (0.119 ml, 1.0 mmol) and a tetrahydrofuran solution of tetrabutyl ammonium fluoride (0.5 ml, 0.5 mmol). After the mixture was stirred for 1 hour at 0° C., it was poured into water and extracted with ether. The ether layer was dried over anhydrous magnesium sulfate and concentrated under vacuum. The resulting reaction mixture was subjected to column chromatography on silica gel and eluted with a 3:7 mixture of ether and hexane to give the titled compound in an amount of 122 mg (yield, 78% based on the starting materials used in Example 6) extracted with ether The ether layer was dried over anhydrous magnesium sulfate concentrated under vacuum The resulting reaction mixture eluted with a 3:7 mixture of ether and hexane

Conditions: temp:4.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: After 1.80 g of Z-Phe was dissolved in 20 mL of MeOH,□73 mg of DMAP and 1.38 g of WSCD HCI were added to the solution at 0° C., followed by stirring at 4° C. for 12 hours. The solvent was concentrated and the concentrate was dissolved in AcOEt. The solution was then washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and then satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated to give Z-Phe-OMe as oil. After dissolving in 20 mL of dry THF, 196 mg of LiBH4 was added to the solution, followed by stirring at room temperature for 15 hours. The solvent was concentrated and the residue was dissolved in AcOEt and the solution washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and then satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated and ether-petroleum ether was added to the concentrate to give 1.45 g (yield 85%) of Z-Phe-ol as the precipitate. After 60 mg of 60% NaH was suspended in 10 mL of dry THF, 285 mg of Z-Phe-ol, 264 mg of 18-crown-6 and 1.48 mL of tert-butyl bromoacetate were added to the solution at 0° C. While reverting to room temperature, the mixture was stirred for 15 hours. After the solvent was distilled off in vacuum, the residue was dissolved in AcOEt and the solution washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated and the concentrate was purified by flash column chromatography to give 217 mg (yield 54%) of Z-PheΨ(CH2O)Gly-OBut as oil. After 160 mg of Z-PheΨ(CH2O)Gly-OBut was dissolved in 20 mL of MeOH, 10% Pd-C was added to the solution, followed by catalytic hydrogenation for 3 hours in a nitrogen flow. The catalyst was removed by filtration and the solvent was concentrated followed by drying. The concentrate was dissolved in 15 mL of DCM, and 114 mg of Fmoc-Cl and 139 μL of DIEA were added to the solution, followed by stirring for 12 hours. After the solvent was distilled off, the residue was dissolved in AcOEt and the solution washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and then satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated and the concentrate was purified by flash column chromatography. Diethyl ether-petroleum ether was added to give 150 mg (yield 77%) of Fmoc-PheΨ(CH2O)Gly-OBut as the precipitate. To the precipitate, were added 31 mg (15 μmol) of H-LeuArg(Me,Boc2)Trp(Boc2)-Rink amide MBHA resin obtained in a manner similar to the process of EXAMPLE 15, 19 mg of Fmoc-PheΨ(CH2O)Gly-OH (prepared by treating Fmoc-PheΨ(CH2O)Gly-OBut with 50% TFA/DCM for an hour), 180 μL of 0.5M HOAt, 42 mg of PyBrop and 47 μL of DIEA. The mixture was shaken for 18 hours. After the resin washed, 5 mL of 20% piperidine/DMF was added to the resin, followed by stirring at room temperature for 30 minutes. After the resin washed, 9 μL of benzoyl chloride, 13 μL of DIEA and 1 mL of DMF 1 were added to the resin, followed by stirring at room temperature for 2 hours. After the resin washed and dried, 200 μL of TFA/PhSMe/m-cresol/TIS/EDT (85/5/5/2.5/2.5) was added to the resin, followed by stirring for 2 hours. Ether was added to the reaction solution, the resulting precipitate was centrifuged and the supernatant was removed. This procedure was repeated for washing. The residue was extracted with an aqueous acetic acid solution and the extract was filtered to remove the resin. Then, linear density gradient elution (60 minutes) was performed with eluants A/B: 75/25-65/35 using: 0.1% TFA in water and eluant B: 0.1% TFA-containing acetonitrile on preparative HPLC using YMC D-ODS-5-ST S-5 120A column (20×150 mm). The fractions containing the product were collected and lyophilized to give 2.5 mg of white powders. The solvent was concentrated the concentrate was dissolved in AcOEt The solution was then washed with 1N HCl aq. solution After drying over Na2SO4 the solvent was concentrated

Conditions: temp:4.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: After 1.80 g of Z-Phe was dissolved in 20 mL of MeOH, 73 mg of DMAP and 1.38 g of WSCD HCl were added to the solution at 0° C., followed by stirring at 4° C. for 12 hours. The solvent was concentrated and the concentrate was dissolved in AcOEt. The solution was then washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and then satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated to give Z-Phe-OMe as oil. After dissolving in 20 mL of dry THF, 196 mg of LiBH4 was added to the solution, followed by stirring at room temperature for 15 hours. The solvent was concentrated and the residue was dissolved in AcOEt and the solution washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and then satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated and ether-petroleum ether was added to the concentrate to give 1.45 g (yield 85%) of Z-Phe-ol as the precipitate. After 60 mg of 60% NaH was suspended in 10 mL of dry THF, 285 mg of Z-Phe-ol, 264 mg of 18-crown-6 and 1.48 mL of tert-butyl bromoacetate were added to the solution at 0° C. While reverting to room temperature, the mixture was stirred for 15 hours. After the solvent was distilled off in vacuum, the residue was dissolved in AcOEt and the solution washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated and the concentrate was purified by flush column chromatography to give 217 mg (yield 54%) of Z-PheΨ(CH2O)Gly-OBut as oil. After 160 mg of Z-PheΨ(CH2O)Gly-OBut was dissolved in 20 mL of MeOH, 10% Pd—C was added to the solution, followed by catalytic hydrogenation for 3 hours in a nitrogen flow. The catalyst was removed by filtration and the solvent was concentrated followed by drying. The concentrate was dissolved in 15 mL of DCM, and 114 mg of Fmoc-Cl and 139 μL of DIEA were added to the solution, followed by stirring for 12 hours. After the solvent was distilled off, the residue was dissolved in AcOEt and the solution washed with 1N HCl aq. solution, satd. NaHCO3 aq. solution and then satd. NaCl aq. solution. After drying over Na2SO4, the solvent was concentrated and the concentrate was purified by flush column chromatography. Diethyl ether-petroleum ether was added to give 150 mg (yield 77%) of Fmoc-PheΨ(CH2O)Gly-OBut as the precipitate. To the precipitate, were added 31 mg (15 μmol) of H-LeuArg(Me,Boc2)Trp(Boc)-Rink amide MBHA resin obtained in a manner similar to the process of EXAMPLE 15, 19 mg of Fmoc-PheΨ(CH2O)Gly-OH (prepared by treating Fmoc-PheΨ(CH2O)Gly-OBut with 50% TFA/DCM for an hour), 180 μL of 0.5M HOAt, 42 mg of PyBrop and 47 μL of DIEA. The mixture was shaken for 18 hours. After the resin washed, 5 mL of 20% piperidine/DMF was added to the resin, followed by stirring at room temperature for 30 minutes. After the resin washed, 9 μL of benzoyl chloride, 13 μL of DIEA and 1 mL of DMF 1 were added to the resin, followed by stirring at room temperature for 2 hours. After the resin washed and dried, 200 μL of TFA/PhSMe/m-cresol/TIS/EDT (85/5/5/2.5/2.5) was added to the resin, followed by stirring for 2 hours. Ether was added to the reaction solution, the resulting precipitate was centrifuged and the supernatant was removed. This procedure was repeated for washing. The residue was extracted with an aqueous acetic acid solution and the extract was filtered to remove the resin. Then, linear density gradient elution (60 minutes) was performed with eluants A/B: 75/25-65/35 using: 0.1% TFA in water and eluant B: 0.1% TFA-containing acetonitrile on preparative HPLC using YMC D-ODS-5-ST S-5 120A column (20×150 mm). The fractions containing the product were collected and lyophilized to give 2.5 mg of white powders. Mass spectrum (M+H)+ 782.2 (Calcd. 782.4) The solvent was concentrated the concentrate was dissolved in AcOEt The solution was then washed with 1N HCl aq. solution After drying over Na2SO4 the solvent was concentrated

Conditions: temp:4.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: After 1.80 g of Z-Phe was dissolved in 20 mL of MeOH, 73 mg of DMAP, 1.38 g of WSCD HCl were added at 0° C., followed by stirring at 4° C. for 12 hours. The solvent was concentrated and the concentrate was dissolved in AcOEt, followed by washing with 1N HCl aq., satd. NaHCO3 aq., and satd. NaCl aq. After drying over Na2SO4, the solvent was concentrated to give Z-Phe-OMe as oil. After dissolving in 20 mL of dry THF, 196 mg of LiBH4 was added thereto and the mixture was stirred at room temperature for 15 hours. The solvent was concentrated and the concentrate was dissolved in AcOEt and the residue was washed with 1N HCl aq., satd. NaHCO3 aq., and satd. NaCl aq. After drying over Na2SO4, the solvent was concentrated and ether-petroleum ether was added to give 1.45 g (yield 85%) of Z-Phe-ol as precipitates. After 60 mg of 60% NaH was suspended in 10 mL of dry THF, 285 mg of Z-Phe-ol, 264 mg of 18-crown-6 and 1.48 mL of tert-butyl bromoacetate were added at 0° C. While returning to room temperature, the mixture was stirred for 15 hours. The solvent was concentrated the concentrate was dissolved in AcOEt by washing with 1N HCl aq. NaCl aq. After drying over Na2SO4 the solvent was concentrated

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

Procedure: 20.0 g (92.73 mmol) of (L)-phenylalanine methyl ester hydrochloride was suspended in 93 ml of toluene, and 15.82 g (92.73 mmols) of benzyl chloroformate was added thereto. 130 ml of an aqueous solution of 1M sodium carbonate was dropwise added thereto, while keeping it at 7° C. or lower, and stirred for 3 hours. After this was subjected to phase separation, the resulting organic layer was washed with 60 ml of 0.1N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate, and then dried with anhydrous sodium sulfate. This was filtered and concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound. The yield was 95.8%. at 7° C. After this was subjected to phase separation the resulting organic layer was washed with 60 ml of 0.1N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate dried with anhydrous sodium sulfate This was filtered concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound

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

Procedure: Under nitrogen gas, diisopropylamine (38.8 g, 383.4 mmol) was added to n-butylmagnesiumchloride (1.8 mol/kg, 178 g, 319.5 mmol) over 30 minutes at 40° C. and the mixture was further stirred for 2 hours at the same temperature to prepare a white slurry (liquor A). Separately, under nitrogen gas was in another vessel, a solution was prepared from methyl (S)-2-benzyloxycarbonylamino-3-phenylpropanoate (20.0 g, 63.9 mmol), dibromomethane (22.22 g, 127.8 mmol) and THF (40 mL) (liquor B). To this liquor B was added said liquor A over 2 hours at an internal temperature of about 5° C., and the reaction was further conducted at 5° C. for 2 hours. This reaction mixture was poured in a mixture of concentrated hydrochloric acid (73.3 g), water (100 g) and ethyl acetate (50 mL) over 1 hour for hydrolysis. After phase separation, the organic layer was washed with 2 portions of water (100 mL each). The organic layer was concentrated under reduced pressure to recover 34.38 g of black oil. This oil was purified by silica gel column chromatography to recover 22.34 g of yellow crude crystals. This crystal crop was recrystallized from ethyl acetate/hexane to provide white crystals of benzyl (S)-(1-benzyl-3,3-dibromo-2-oxopropyl)carbamate (11.99 g, 88.0 area %, yield 57%). to prepare a white slurry (liquor A) Separately, under nitrogen gas was in another vessel To this liquor B was added over 2 hours at an internal temperature of about 5° C. the reaction was further conducted at 5° C. for 2 hours After phase separation the organic layer was washed with 2 portions of water (100 mL each) The organic layer was concentrated under reduced pressure to recover 34.38 g of black oil This oil was purified by silica gel column chromatography to recover 22.34 g of yellow crude crystals This crystal crop was recrystallized from ethyl acetate/hexane

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

Procedure: N-Benzyloxycarbonyl phenylalanine methyl ester is reduced with DIBAL to the aldehyde using the procedure of Boger et al., J. Med. Chem. (1985) 28, 1779, which is homologated with the sodio derivative of trimethyl phosphonoacetate using the procedure of Wadsworth and Emmons, Org. Synth. (1965) 45, 44 to give methyl 4-(benzyloxycarbonyl)amino-5-phenyl-2-pentenoate.

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

Procedure: 20.0 g (92.73 mmol) of (L)-phenylalanine methyl ester hydrochloride was suspended in 93 ml of toluene, and 15.82 g (92.73 mmols) of benzyl chloroformate was added thereto. 130 ml of an aqueous solution of 1 M sodium carbonate was dropwise added thereto, while keeping it at 7° C. or lower, and stirred for 3 hours. After this was subjected to phase separation, the resulting organic layer was washed with 60 ml of 0.1 N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate, and then dried with anhydrous sodium sulfate. This was filtered and concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound. The yield was 95.8%. at 7° C. After this was subjected to phase separation the resulting organic layer was washed with 60 ml of 0.1 N hydrochloric acid and 60 ml of a saturated aqueous solution of sodium hydrogen carbonate dried with anhydrous sodium sulfate This was filtered concentrated to obtain 28.75 g (96.8 wt. %, 88.81 mmol) of the above-entitled compound