O-SUBSTITUTED SERINE DERIVATIVE PRODUCTION METHOD

Abstract

It was discovered that a cyclic sulfamidate can be produced by reacting an amino acid derivative with a cyclization reagent. In addition, it was discovered that an O-substituted serine derivative can be produced by reacting a cyclic sulfamidate with an alcohol.

Claims

1. A method for producing a compound represented by general formula (I): ##STR00126## [wherein, R.sub.1 is optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.8 cycloalkyl, optionally substituted aralkyl, or optionally substituted heteroaralkyl, R.sub.2 is C.sub.1-C.sub.6 alkyl or an amino protecting group, R.sub.4 is a carboxyl protecting group, L.sub.1 is a single bond or —CH.sub.2—, L.sub.2 is a single bond or —CH.sub.2—, and n is 1 or 2, with the proviso that when L.sub.1 is —CH.sub.2—, L.sub.2 is a single bond, and when L.sub.2 is —CH.sub.2—, L.sub.1 is a single bond], its chemically acceptable salt, or a solvate thereof, the method comprising the following steps: Step A: reacting a compound represented by general formula (V): ##STR00127## [wherein, R.sub.2, R.sub.4, L.sub.1, L.sub.2, and n are synonymous with those described above], its chemically acceptable salt, or a solvate thereof with a cyclization reagent to obtain a compound represented by general formula (IV): ##STR00128## [wherein, R.sub.2, R.sub.4, L.sub.1, L.sub.2, and n are synonymous with those described above], its chemically acceptable salt, or a solvate thereof, Step B: reacting the compound represented by general formula (IV), its chemically acceptable salt, or a solvate thereof with an oxidizing agent to obtain a compound represented by general formula (II): ##STR00129## [wherein, R.sub.2, R.sub.4, L.sub.1, L.sub.2, and n are synonymous with those described above], its chemically acceptable salt, or a solvate thereof, and Step C: reacting the compound represented by general formula (II), its chemically acceptable salt, or a solvate thereof with R.sub.1OH (wherein, R.sub.1 is synonymous with those described above) to obtain the compound represented by general formula (I), its chemically acceptable salt, or a solvate thereof.

2. A method for producing a compound represented by general formula (I): ##STR00130## [wherein, R.sub.1 is optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted C.sub.3-C.sub.8 cycloalkyl, optionally substituted aralkyl, or optionally substituted heteroaralkyl, R.sub.2 is a hydrogen, R.sub.4 is a carboxyl protecting group, L.sub.1 is a single bond or —CH.sub.2—, L.sub.2 is a single bond or —CH.sub.2—, and n is 1 or 2, with the proviso that when L.sub.1 is —CH.sub.2—, L.sub.2 is a single bond, and when L.sub.2 is —CH.sub.2—, L.sub.1 is a single bond], its chemically acceptable salt, or a solvate thereof, the method comprising the following steps: Step A: reacting a compound represented by general formula (V′): ##STR00131## [wherein, R.sub.2′ is an amino protecting group, and R.sub.4, L.sub.1, L.sub.2, and n are synonymous with those described above], its chemically acceptable salt, or a solvate thereof with a cyclization reagent to obtain a compound represented by general formula (IV′): ##STR00132## [wherein, R.sub.2′, R.sub.4, L.sub.1, L.sub.2, and n are synonymous with those described above], its chemically acceptable salt, or a solvate thereof, Step B: reacting the compound represented by general formula (IV), its chemically acceptable salt, or a solvate thereof with an oxidizing agent to obtain a compound represented by general formula (II′): ##STR00133## [wherein, R.sub.2′, R.sub.4, L.sub.1, L.sub.2, and n are synonymous with those described above], its chemically acceptable salt, or a solvate thereof, and Step C: reacting the compound represented by general formula (II′), its chemically acceptable salt, or a solvate thereof with R.sub.1OH (wherein, R.sub.1 is synonymous with that described above) to obtain the compound represented by general formula (I), its chemically acceptable salt, or a solvate thereof.

3. The method of claim 1 or 2, further comprising a step (Step D) of deprotecting the carboxyl protecting group represented by R.sub.4 of the compound represented by general formula (I), its chemically acceptable salt, or a solvate thereof to obtain a compound represented by general formula (I′): ##STR00134## [wherein, R.sub.1, R.sub.2, L.sub.1, L.sub.2, and n are synonymous with those in claim 1 or 2], its chemically acceptable salt, or a solvate thereof.

4. The method of claim 3, further comprising a step (Step E) of introducing a group represented by R.sub.3 to the amino group of the compound represented by general formula (I′), its chemically acceptable salt, or a solvate thereof to obtain a compound represented by general formula (I″): ##STR00135## [wherein, R.sub.1, R.sub.2, L.sub.1, L.sub.2, and n are synonymous with those in claim 1 or 2, and R.sub.3 is an amino protecting group or C.sub.1-C.sub.4 alkyl], its chemically acceptable salt, or a solvate thereof.

5. The method of claim 1, 3, or 4, wherein R.sub.1 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, aralkyl, or heteroaralkyl, which is optionally substituted with one or more substituents independently selected from halogen, aryl that is optionally substituted with halogen, or hydroxyl, R.sub.2 is selected from Boc, Fmoc, Cbz, or Alloc, and R.sub.4 is benzyl or tert-Bu.

6. The method of any one of claims 2 to 4, wherein R.sub.1 is C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.8 cycloalkyl, aralkyl, or heteroaralkyl, which is optionally substituted with one or more substituents independently selected from halogen, aryl that is optionally substituted with halogen, or hydroxyl, R.sub.2′ is selected from Boc, Fmoc, Cbz, or Alloc, and R.sub.4 is benzyl or tert-Bu.

7. The method of claim 4, wherein R.sub.3 is selected from Boc, Fmoc, Cbz, Alloc, or methyl.

8. The method of any one of claims 1 to 7, wherein the oxidizing agent used in Step B is a combination of periodate and ruthenium catalyst.

9. The method of claim 8, wherein periodate that is 1.5 to 5 equivalents and ruthenium catalyst that is 0.01 to 0.2 equivalents to the compound represented by general formula (IV), its chemically acceptable salt, or a solvate thereof, or to the compound represented by general formula (IV′), its chemically acceptable salt, or a solvate thereof, are used.

10. The method of any one of claims 1 to 9, wherein Step B is performed in a solvent mixture of acetonitrile and water.

11. The method of any one of claims 1 to 10, wherein Step C is performed in the presence of an acid salt.

12. The method of any one of claims 1 to 11, wherein Step C is performed in 2,2,2-trifluoroethanol, 1,1,1,3,3,3-hexafluoro-2-propanol, or 2-methyltetrahydrofuran.

13. The method of any one of claims 1 to 12, wherein Step C further comprises a step of extracting the reaction mixture with an organic solvent, and the extract is used in Step D without concentration to dryness.

14. The method of any one of claims 3 to 13, wherein Step D is performed in the presence of a Pd catalyst, and/or in the presence of hydrogen gas, formic acid, or ammonium formate.

15. The method of any one of claims 1 to 14, wherein Step A is performed in ethyl acetate, isopropyl acetate, or butyl acetate, and thionyl chloride that is 1.5 to 5 equivalents to the compound represented by general formula (V), its chemically acceptable salt, or a solvate thereof, or to the compound represented by general formula (V′), its chemically acceptable salt, or a solvate thereof is used.

Description

EXAMPLE

[0257] Herein below, the present invention is explained in further detail with reference to Examples, but it is not to be construed as being limited thereto.

[0258] Condition 1 for High Performance Liquid Chromatography [0259] Equipment: UPLC ACQUITY, Waters Corporation; [0260] Column: BEH (1.7 μm, 2.1 mm I.D.×50 mm, Waters Corporation); [0261] Mobile phase: water containing 0.05% trifluoroacetic acid (A) and acetonitrile containing 0.05% trifluoroacetic acid (B); [0262] Elution: stepwise solvent gradient elution with 5% B to 100% B (4.0 min.), hold at 100% B (0.5 min.); [0263] Flow rate: 0.5 mL/min. [0264] Column temperature: 35° C.

[0265] Condition 2 for High Performance Liquid Chromatography [0266] Equipment: UPLC ACQUITY, Waters Corporation; [0267] Column: BEH (1.7 μm, 2.1 mm I.D.×50 mm, Waters Corporation); [0268] Mobile phase: water containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B); [0269] Elution: stepwise solvent gradient elution with 5% B to 100% B (4.0 min.), hold at 100% B (0.5 min.); [0270] Flow rate: 0.5 mL/min. [0271] Column temperature: 25° C.

[0272] Condition 3 for High Performance Liquid Chromatography [0273] Equipment: UPLC ACQUITY, Waters Corporation; [0274] Column: CHIRALCELL OD-3R (3.0 μm, 4.6 mm I.D.×50 mm, Daicel Corporation); Mobile phase: water containing 0.1% formic acid (A) and acetonitrile containing 0.1% formic acid (B); [0275] Elution: stepwise solvent gradient elution with 5% B to 100% B (4.0 min.), hold at 100% B (0.5 min.); [0276] Flow rate: 1.5 mL/min. [0277] Column temp.: 25° C.

[0278] Condition 4 for High Performance Liquid Chromatography [0279] Equipment: UPLC ACQUITY, Waters Corporation; [0280] Column: CHIRALPAK IA-3 (3.0 μm, 4.6 mm I.D.×50 mm, Daicel Corporation); [0281] Mobile phase: water containing 10 mM ammonium acetate (A) and methanol containing 10 mM ammonium acetate (B); [0282] Elution: stepwise solvent gradient elution with 5% B to 60% B (0.5 min.), 60% B to 80% B (3.0 min.), 80% B to 100% B (0.5 min.), hold at 100% B (0.5 min.); [0283] Flow rate: 1.2 mL/min. [0284] Column temperature: 25° C.

[0285] Condition 5 for High Performance Liquid Chromatography [0286] Equipment: UPLC ACQUITY, Waters Corporation; [0287] Column: CHIRALPAK IG-3 (3.0 μm, 4.6 mm I.D.×50 mm, Daicel Corporation); [0288] Mobile phase: water containing 10 mM ammonium acetate (A) and methanol containing 10 mM ammonium acetate (B); [0289] Elution: stepwise solvent gradient elution with 5% B to 60% B (0.1 min.), 60% B to 100% B (3.4 min.), hold at 100% B (1.0 min.); [0290] Flow rate: 1.2 mL/min. [0291] Column temperature: 25° C.

[0292] .sup.1H-NMR spectrum was measured using AVANCE III HD 400 BBFO-SMART probe (Bruker corporation), where the chemical shift of Me.sub.4Si used as an internal standard was set to be 0 ppm, and the deuterium lock signal from the sample solvent was used as a reference. The chemical shifts of the signals of the analyte compound were expressed in ppm. The abbreviations for signal splitting were: s=singlet, brs=broad singlet, d=doublet, t=triplet, q=quartet, dd=double doublet, and m=multiplet, and the separation in a split signal was expressed in J (Hz). The integrated value of a signal was calculated based on the ratio of signal area intensity of the signals. Production example using Boc-Ser-OBzl as a starting material.

Example 1: benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidate-4-carboxylate

[0293] ##STR00026##

[0294] 1) To a solution consisting of thionyl chloride 16.1 g (135 mmol) and ethyl acetate 400 mL cooled to −15° C., a solution consisting of Boc-Ser-OBzl 20 g (68 mmol) and ethyl acetate 50 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then pyridine 26.8 g (338 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 15 minutes, and then at room temperature for 22 hours. To the reaction mixture, water 200 mL was added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 1N-hydrochloric acid 200 mL and 10% brine 200 mL, and concentrated under reduced pressure to give a crude product of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamiditecarboxylate 23.9 g as a diastereomeric mixture.

[0295] 2) To a solution consisting of the crude product of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamiditecarboxylate 23.9 g and acetonitrile 100 mL cooled to 0° C., a solution consisting of sodium periodate 21.7 g (101 mmol), ruthenium chloride hydrate 0.14 g (0.68 mmol), and water 300 mL was added dropwise over 8 minutes. The reaction mixture was stirred at the same temperature for 22 minutes, and then at room temperature for 1 hour. To the reaction mixture, 10% aqueous sodium carbonate 60 mL, water 100 mL, and ethyl acetate 220 mL were added, and the organic layer and the aqueous layer were separated. To the aqueous layer obtained, ethyl acetate 100 mL was added again, and the organic layer and the aqueous layer were separated. The organic layers obtained were combined, washed with 10% brine 200 mL, and concentrated under reduced pressure to give benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 22.3 g as a crude product.

[0296] 3) To a mixture consisting of the crude product of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 22.3 g and ethyl acetate 70 mL heated to 50° C., hexane 280 mL was added, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 2 hours, and the precipitate was collected by filtration under reduced pressure. The crystals obtained were dried under reduced pressure to give benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 19.8 g (yield: 81.6% in 2 steps) as white crystals.

Benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidate-4-carboxylate

[0297] ##STR00027##

[0298] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.87 min., condition 3 for high performance liquid chromatography) UV intensity ratio: 99.2% (detection wavelength 205 nm, retention time 2.77 min., condition 2 for high performance liquid chromatography)

[0299] .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ: 1.50 (9H, s), 4.67 (1H, dd, J=9.6, 2.0), 4.76 (1H, dd, J=9.6, 6.4), 4.80-4.86 (1H, m), 5.23 (1H, d, J=12.0), 5.32 (1H, d, J=12.0), 7.30-7.42 (5H, m)

Example 2: Fmoc-Ser(n-Pr)—OH

[0300] ##STR00028##

[0301] 1) A mixture consisting of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 6.00 g (16.7 mmol) and 1-propanol 120 mL was stirred for 12 hours while heating at 90° C. To the reaction mixture, ethyl acetate 240 mL and 5% aqueous sodium hydrogen carbonate 240 mL were added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 10% brine 240 mL to give H-Ser(n-Pr)-OBzl as an ethyl acetate solution.

H-Ser(n-Pr)-OBzl

[0302] ##STR00029##

[0303] Optical purity: 99.5% ee (detection wavelength 205 nm, retention time 2.76 min., condition 4 for high performance liquid chromatography)

[0304] UV intensity ratio: 87.3% (detection wavelength 205 nm, retention time 1.42 min., condition 2 for high performance liquid chromatography)

[0305] 2) To a solution of H-Ser(n-Pr)-OBzl in ethyl acetate, a mixture of 10% palladium on carbon 0.6 g and methanol 30 mL was added, and the reaction mixture was stirred under hydrogen gas atmosphere at room temperature for 2 hours. Palladium catalyst was filtered out using Celite under reduced pressure, and the mixture obtained was concentrated under reduced pressure to give H-Ser(n-Pr)—OH 7.82 g as a crude product.

[0306] 3) To a solution consisting of H-Ser(n-Pr)—OH 7.82 g, water 96 mL, and sodium carbonate 4.80 g (45.2 mmol) cooled to 0° C., a solution consisting of FmocOSu 4.00 g (11.8 mmol) and acetonitrile 96 mL was added dropwise over 5 minutes. The reaction mixture was stirred at room temperature for 24 hours, 2N-hydrochloric acid 48 mL and water 48 mL were added thereto over 10 minutes and after a precipitate appeared, the reaction mixture was stirred at room temperature for 2 hours. To the reaction mixture, water 24 mL was added, and the mixture was stirred for 1 hour, additional water 24 mL was added, and the mixture was stirred for 2 hours, and then the precipitate was collected by filtration under reduced pressure. The crystals obtained were dried under reduced pressure to give Fmoc-Ser(n-Pr)—OH 3.60 g (yield: 58.4% in 3 steps) as white crystals.

Fmoc-Ser(n-Pr)—OH

[0307] ##STR00030##

[0308] Optical purity: 99.6% ee (detection wavelength 205 nm, retention time 3.35 min., condition 3 for high performance liquid chromatography) UV intensity ratio: 96.2% (detection wavelength 205 nm, retention time 2.74 min., condition 2 for high performance liquid chromatography)

[0309] .sup.1H-NMR (DMSO-d6, 400 MHz) δ: 0.84 (3H, t, J=7.2), 1.42-1.56 (2H, m), 3.28-3.42 (2H, m), 3.56-3.70 (2H, m), 4.16-4.34 (4H, m), 7.32 (2H, dt, J=7.2, 0.8), 7.42 (2H, t, J=7.6), 7.61 (1H, d, J=8.0), 7.74 (2H, d, J=7.8), 7.89 (2H, d, J=7.6), 12.76 (1H, brs) yy

Example 3: Fmoc-Ser(i-Pr)—OH

[0310] ##STR00031##

[0311] 1) A mixture consisting of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 1.00 g (2.8 mmol) and 2-propanol 20 mL was stirred for 36 hours while heating at 80° C. To the reaction mixture, ethyl acetate 20 mL, 5% aqueous sodium hydrogen carbonate 20 mL, and 10% brine 20 mL were added, and the organic layer and the aqueous layer were separated. The organic layer was obtained as an ethyl acetate solution of H-Ser(i-Pr)-OBzl.

H-Ser(i-Pr)-OBzl

[0312] ##STR00032##

[0313] Optical purity: 99.9% ee (detection wavelength 205 nm, condition 4 for high performance liquid chromatography)

[0314] UV intensity ratio: 84.4% (detection wavelength 205 nm, retention time 1.38 min., condition 2 for high performance liquid chromatography)

[0315] 2) To a solution of H-Ser(i-Pr)-OBzl in ethyl acetate, a mixture of 10% palladium on carbon 0.15 g and methanol 5 mL was added, and the reaction mixture was stirred under hydrogen gas atmosphere at room temperature for 2 hours. Palladium catalyst was filtered out using Celite under reduced pressure, and the mixture obtained was concentrated under reduced pressure to give H-Ser(i-Pr)—OH 842 mg as a crude product.

[0316] 3) To a solution consisting of H-Ser(i-Pr)—OH 842 mg, water 6 mL, and sodium carbonate 0.31 g (2.9 mmol) cooled to 0° C., a solution consisting of FmocOSu 0.66 g (2.0 mmol) and acetonitrile 6 mL was added dropwise over 2 minutes. The reaction mixture was stirred at room temperature for 3 hours, 1N-hydrochloric acid 10 mL was added thereto over 5 minutes, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 3 hours. After stirring the reaction mixture for 3 hours, the precipitate was collected by filtration under reduced pressure. The crystals obtained were dried under reduced pressure to give Fmoc-Ser(i-Pr)—OH 667 mg (yield: 63.5% in 3 steps) as white crystals.

Fmoc-Ser(i-Pr)—OH

[0317] ##STR00033##

[0318] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 3.25 min., condition 3 for high performance liquid chromatography)

[0319] UV intensity ratio: 98.7% (detection wavelength 205 nm, retention time 2.74 min., condition 2 for high performance liquid chromatography)

[0320] .sup.1H-NMR (DMSO-d6, 400 MHz) δ: 1.06 (3H, d, J=6.0), 1.08 (3H, d, J=6.0), 3.48-3.66 (3H, m), 4.10-4.32 (4H, m), 7.33 (2H, dt, J=7.6, 0.8), 7.42 (2H, t, J=7.0), 7.56 (1H, d, J=8.0), 7.74 (2H, d, J=7.8), 7.89 (2H, d, J=7.6), 12.73 (1H, brs)

Example 4: H-Ser(2-hydroxy-2-methylpropyl)-OBzl

[0321] ##STR00034##

[0322] A mixture consisting of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.10 mmol) and 2-methylpropane-1,2-diol 1.00 mL was stirred for 30 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

H-Ser(2-hydroxy-2-methylpropyl)-OBzl

[0323] ##STR00035##

[0324] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 1.78 min., condition 4 for high performance liquid chromatography)

[0325] UV intensity ratio: 73.3% (detection wavelength 205 nm, retention time 1.22 min., condition 2 for high performance liquid chromatography)

Example 5: H-Ser(n-Bu)-OBzl

[0326] ##STR00036##

[0327] A mixture consisting of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 25 mg (0.07 mmol) and 1-butanol 0.50 mL was stirred for 24 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

H-Ser(n-Bu)-OBzl

[0328] ##STR00037##

[0329] UV intensity ratio: 77.0% (detection wavelength 205 nm, retention time 1.83 min., condition 1 for high performance liquid chromatography)

[0330] ESI (LC/MS positive mode) m/z: 252.48 (M+H.sup.+)

Example 6: H-Ser(3-methylbutyl)-OBzl

[0331] ##STR00038##

[0332] A mixture consisting of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 25 mg (0.07 mmol) and 3-methylbutanol 0.50 mL was stirred for 24 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

H-Ser(3-methylbutyl)-OBzl

[0333] ##STR00039##

[0334] UV intensity ratio: 73.9% (detection wavelength 205 nm, retention time 2.00 min., condition 1 for high performance liquid chromatography)

[0335] ESI (LC/MS positive mode) m/z: 266.52 (M+H.sup.+)

Example 7: Benzyl (4S)-1,2,5-sulfamidate-4-carboxylate

[0336] ##STR00040##

[0337] A mixture consisting of benzyl (4S)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 1.00 g (2.8 mmol) and 2,2,2-trifluoroethanol 10 mL was stirred for 4 hours while heating at 70° C. To the reaction mixture, ethyl acetate 20 mL and 5% brine 40 mL were added, and the organic layer and the aqueous layer were separated. The organic layer was concentrated under reduced pressure to give benzyl (4S)-1,2,5-sulfamidatecarboxylate 734 mg as a crude product.

[0338] The crude product obtained was purified with silica gel column chromatography (elution solvent: ethyl acetate-hexane) to give benzyl (4S)-1,2,5-sulfamidatecarboxylate 649 mg (yield: 90.2%) as a pale yellow solid.

Benzyl (4S)-1,2,5-sulfamidatecarboxylate

[0339] ##STR00041##

[0340] UV intensity ratio: 99.6% (detection wavelength 205 nm, retention time 1.94 min., condition 1 for high performance liquid chromatography)

[0341] .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ: 4.44-4.52 (1H, m), 4.56 (1H, dd, J=8.8, 5.6), 4.74 (1H, dd, J=8.8, 7.6), 5.09-5.18 (1H, m), 5.27 (1H, d, J=11.6), 5.30 (1H, d, J=11.6), 7.32-7.44 (5H, m)

[0342] Production example using D-Boc-Ser-OBzl as a starting material

Example 8: Benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidate-4-carboxylate

[0343] ##STR00042##

[0344] 1) To a solution consisting of thionyl chloride 8.05 g (67.7 mmol) and acetonitrile 140 mL cooled to −40° C., a solution consisting of D-Boc-Ser-OBzl 10.0 g (33.8 mmol) and acetonitrile 30 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then pyridine 13.4 g (169 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then at room temperature for 4 hours. To the reaction mixture, water 340 mL and ethyl acetate 170 mL were added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 5% aqueous sodium hydrogen carbonate 170 mL, and with a mixed solution of 0.5N-hydrochloric acid 170 mL and 10% brine 170 mL, and concentrated under reduced pressure to give a crude product of benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamiditecarboxylate 10.76 g as a diastereomeric mixture.

[0345] 2) To a solution consisting of the crude product of benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamiditecarboxylate 10.76 g and acetonitrile 160 mL cooled to 0° C., a solution consisting of sodium periodate 10.7 g (50 mmol), ruthenium chloride hydrate 62 mg (0.30 mmol), and water 160 mL was added dropwise over 15 minutes. The reaction mixture was stirred at the same temperature for 2 hours. To the reaction mixture, 5% aqueous sodium hydrogen carbonate 160 mL and ethyl acetate 160 mL were added, and the organic layer and the aqueous layer were separated. To the aqueous layer obtained, ethyl acetate 160 mL was added again, and the organic layer and the aqueous layer were separated. The organic layers obtained were combined, washed with 10% brine 160 mL, and concentrated under reduced pressure to give benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 10.44 g as a crude product.

[0346] 3) To a mixture consisting of the crude product of benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 10.44 g and ethyl acetate 30 mL heated to 40° C., hexane 120 mL was added, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 2 hours, and the precipitate was collected by filtration under reduced pressure. The crystals obtained were dried under reduced pressure to give benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 9.00 g (yield: 74.5% in 2 steps) as white crystals.

Benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate

[0347] ##STR00043##

[0348] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.93 min., condition 3 for high performance liquid chromatography)

[0349] UV intensity ratio: 97.7% (detection wavelength 205 nm, retention time 2.77 min., condition 2 for high performance liquid chromatography)

[0350] .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ: 1.49 (9H, s), 4.67 (1H, dd, J=9.6, 2.2), 4.76 (1H, dd, J=9.6, 6.4), 4.80-4.86 (1H, m), 5.23 (1H, d, J=12.0), 5.32 (1H, d, J=12.0), 7.30-7.42 (5H, m)

Example 9: Racemization of D-H-Ser(n-Pr)-OBzl

[0351] ##STR00044##

[0352] 1) Reaction mixture: A mixture consisting of benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 1.00 g (2.8 mmol) and 1-propanol 20 mL was stirred for 15 hours while heating at 90° C., and the reaction mixture was analyzed with HPLC.

D-H-Ser(n-Pr)-OBzl

[0353] ##STR00045##

[0354] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.54 min., condition 4 for high performance liquid chromatography)

[0355] UV intensity ratio: 82.1% (detection wavelength 205 nm, retention time 1.40 min., condition 2 for high performance liquid chromatography)

[0356] 2) Ethyl acetate solution 1: To the reaction mixture, ethyl acetate 40 mL and 5% aqueous sodium hydrogen carbonate 40 mL were added, and the organic layer and the aqueous layer were separated. The organic layer obtained was analyzed with HPLC.

[0357] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.55 min., condition 4 for high performance liquid chromatography)

[0358] UV intensity ratio: 84.7% (detection wavelength 205 nm, retention time 1.42 min., condition 2 for high performance liquid chromatography)

[0359] 3) Ethyl acetate solution 2: The ethyl acetate solution 1 obtained was divided into two, and one part of them was washed twice with 10% brine 20 mL, and the organic layer obtained was analyzed with HPLC.

[0360] Optical purity: 99.8% ee (detection wavelength 205 nm, retention time 2.55 min., condition 4 for high performance liquid chromatography)

[0361] UV intensity ratio: 85.1% (detection wavelength 205 nm, retention time 1.42 min., condition 2 for high performance liquid chromatography)

[0362] 4) Crude product 1: The ethyl acetate solution 2 obtained was divided into two, and one part of them was concentrated under reduced pressure in a water bath set at 25° C., and the crude product of D-H-Ser(n-Pr)-OBzl obtained was analyzed with HPLC.

[0363] Optical purity: 82.8% ee (detection wavelength 205 nm, retention time 2.54 min., condition 4 for high performance liquid chromatography)

[0364] UV intensity ratio: 79.7% (detection wavelength 205 nm, retention time 1.39 min., condition 2 for high performance liquid chromatography)

[0365] 5) Crude product 2: The other part of the organic layer that was divided into two was concentrated under reduced pressure in a water bath set at 50° C., and the crude product of D-H-Ser(n-Pr)-OBzl obtained was analyzed with HPLC.

[0366] Optical purity: 72.6% ee (detection wavelength 205 nm, retention time 2.53 min., condition 4 for high performance liquid chromatography)

[0367] UV intensity ratio: 79.1% (detection wavelength 205 nm, retention time 1.39 min., condition 2 for high performance liquid chromatography)

Example 10: Stability of the Optical Purity of D-H-Ser(n-Pr)-OBzl

[0368] 1) Two types of ethyl acetate solution prepared in Example 9 were left to stand at room temperature for 3 days, and were analyzed with HPLC.

[0369] Ethyl Acetate Solution 1:

[0370] Optical purity: 91.5% ee (detection wavelength 205 nm, retention time 2.54 min., condition 4 for high performance liquid chromatography)

[0371] Ethyl Acetate Solution 2:

[0372] Optical purity: 94.7% ee (detection wavelength 205 nm, retention time 2.54 min., condition 4 for high performance liquid chromatography)

[0373] 2) Two types of ethyl acetate solution prepared in Example 9 were left to stand at room temperature for 5 days, and were analyzed with HPLC.

[0374] Ethyl Acetate Solution 1:

[0375] Optical purity: 85.6% ee (detection wavelength 205 nm, retention time 2.56 min., condition 4 for high performance liquid chromatography)

[0376] Ethyl Acetate Solution 2:

[0377] Optical purity: 88.9% ee (detection wavelength 205 nm, retention time 2.55 min., condition 4 for high performance liquid chromatography)

[0378] 3) Two types of ethyl acetate solution prepared in Example 9 were left to stand at room temperature for 10 days, and were analyzed with HPLC.

[0379] Ethyl Acetate Solution 1:

[0380] Optical purity: 76.8% ee (detection wavelength 205 nm, retention time 2.55 min., condition 4 for high performance liquid chromatography)

[0381] Ethyl Acetate Solution 2:

[0382] Optical purity: 82.9% ee (detection wavelength 205 nm, retention time 2.54 min., condition 4 for high performance liquid chromatography)

Example 11: D-Fmoc-Ser(n-Pr)—OH

[0383] ##STR00046##

[0384] 1) To the other part of the ethyl acetate solution 1, which was produced in Example 9 and divided into two, a mixture of 10% palladium on carbon 0.15 g and methanol 5 mL was added, and the reaction mixture was stirred under hydrogen gas atmosphere at room temperature for 2 hours. Palladium catalyst was filtered out using Celite under reduced pressure, and the mixture obtained was concentrated under reduced pressure to give D-H-Ser(n-Pr)—OH 854 mg as a crude product.

[0385] 2) To a solution consisting of D-H-Ser(n-Pr)—OH 845 mg, water 8 mL, and sodium carbonate 0.40 g (3.77 mmol) cooled to 0° C., a solution consisting of FmocOSu 337 mg (1.00 mmol) and acetonitrile 8 mL was added dropwise over 5 minutes. The reaction mixture was stirred at room temperature for 3 hours, 1N-hydrochloric acid 8 mL was added thereto, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 1 hour and 20 minutes. To the reaction mixture, 0.1N-hydrochloric acid 2 mL was added, and the mixture was stirred for additional 1 hour and 20 minutes, and then the precipitate was collected by filtration under reduced pressure. The crystals obtained were dried under reduced pressure to give D-Fmoc-Ser(n-Pr)—OH 3.60 g (yield: 58.4% in 3 steps) as white crystals.

D-Fmoc-Ser(n-Pr)—OH

[0386] ##STR00047##

[0387] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 3.12 min., condition 3 for high performance liquid chromatography)

[0388] UV intensity ratio: 97.3% (detection wavelength 205 nm, retention time 2.74 min., condition 2 for high performance liquid chromatography)

[0389] .sup.1H-NMR (DMSO-d6, 400 MHz) δ: 0.84 (3H, t, J=7.2), 1.42-1.56 (2H, m), 3.28-3.42 (2H, m), 3.56-3.70 (2H, m), 4.16-4.34 (4H, m), 7.32 (2H, dt, J=7.2, 0.8), 7.42 (2H, t, J=7.6), 7.61 (1H, d, J=8.0), 7.74 (2H, d, J=7.8), 7.89 (2H, d, J=7.6), 12.76 (1H, brs)

Example 12: D-Fmoc-Ser(i-Pr)—OH

[0390] ##STR00048##

[0391] 1) A mixture consisting of benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.14 mmol) and 2-propanol 1 mL was stirred for 30 hours while heating at 80° C. To the reaction mixture, ethyl acetate 1 mL, 5% aqueous sodium hydrogen carbonate 0.5 mL, and 10% brine 0.5 mL were added, and the organic layer and the aqueous layer were separated. The organic layer was obtained as an ethyl acetate solution of D-H-Ser(i-Pr)-OBzl.

D-H-Ser(i-Pr)-OBzl

[0392] ##STR00049##

[0393] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.21 min., condition 4 for high performance liquid chromatography)

[0394] UV intensity ratio: 78.5% (detection wavelength 205 nm, retention time 1.36 min., condition 2 for high performance liquid chromatography)

[0395] 2) To a solution of D-H-Ser(i-Pr)-OBzl in ethyl acetate, a mixture of 10% palladium on carbon 15 mg and methanol 1 mL was added, and the reaction mixture was stirred under hydrogen gas atmosphere at room temperature for 1 hour. Palladium catalyst was filtered out using Celite under reduced pressure, and the mixture obtained was concentrated under reduced pressure to give D-H-Ser(i-Pr)—OH 27 mg as a crude product.

[0396] 3) To a solution consisting of D-H-Ser(i-Pr)—OH 27 mg, water 0.5 mL, and sodium carbonate 25 mg (0.235 mmol) cooled to 0° C., a solution consisting of FmocOSu 33 mg (0.098 mmol) and acetonitrile 0.5 mL was added dropwise over 2 minutes. The reaction mixture was stirred at room temperature for 5 hours, 1N-hydrochloric acid 10 mL was added thereto over 5 minutes, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 2 hours, and analyzed with HPLC.

D-Fmoc-Ser(i-Pr)—OH

[0397] ##STR00050##

[0398] Optical purity: >95% ee, the exact value was unknown because the optical isomer and impurities could not be separated (detection wavelength 205 nm, retention time 3.05 min., condition 3 for high performance liquid chromatography)

[0399] UV intensity ratio: 85.6% (detection wavelength 205 nm, retention time 2.70 min., condition 2 for high performance liquid chromatography)

Example 13: D-H-Ser(2-hydroxy-2-methylpropyl)-OBzl

[0400] ##STR00051##

[0401] A mixture consisting of benzyl (4R)-5-t-butoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.14 mmol) and 2-methylpropane-1,2-diol 1 mL was stirred for 30 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

D-H-Ser(2-hydroxy-2-methylpropyl)-OBzl

[0402] ##STR00052##

[0403] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.16 min., condition 4 for high performance liquid chromatography)

[0404] UV intensity ratio: 73.4% (detection wavelength 205 nm, retention time 1.22 min., condition 2 for high performance liquid chromatography)

[0405] Production example using Fmoc-Ser-OBzl as a starting material

Example 14: benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-carboxylate

[0406] ##STR00053##

[0407] 1) To a solution consisting of thionyl chloride 14.3 g (120 mmol) and ethyl acetate 350 mL cooled to −15° C., a solution consisting of Fmoc-Ser-OBzl 25 g (60 mmol) and ethyl acetate 100 mL was added dropwise over 10 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then pyridine 23.7 g (300 mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then at room temperature for 24 hours. To the reaction mixture, water 200 mL was added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 1N-hydrochloric acid 200 mL and 10% brine 200 mL, and concentrated under reduced pressure to give a crude product of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamiditecarboxylate 28.0 g as a diastereomeric mixture.

[0408] 2) To a solution consisting of the crude product of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamiditecarboxylate 28.0 g and acetonitrile 200 mL cooled to −10° C., a solution consisting of sodium periodate 19.3 g (90 mmol), ruthenium chloride hydrate 0.12 g (0.6 mmol), and water 300 mL was added dropwise over 15 minutes. The reaction mixture was stirred at the same temperature for 15 minutes, and then at room temperature for 23 hours. To the reaction mixture, water 100 mL and ethyl acetate 300 mL were added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 10% brine 200 mL, and concentrated under reduced pressure to give benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 28.7 g as a crude product.

[0409] 3) The crude product obtained was purified with silica gel column chromatography (elution solvent: ethyl acetate-hexane) to give benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 15.2 g (yield: 53.0% in 2 steps) as a white powder.

Benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-carboxylate

[0410] ##STR00054##

[0411] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 4.02 min., condition 3 for high performance liquid chromatography)

[0412] UV intensity ratio: 95.2% (detection wavelength 205 nm, retention time 3.31 min., condition 2 for high performance liquid chromatography)

[0413] 1H-NMR (CDCl3, 400 MHz) δ: 4.22-4.34 (1H, m), 4.46 (1H, dd, J=10.4, 7.2), 4.58 (1H, dd, J=10.4, 7.2), 4.70-4.96 (3H, m), 5.20-5.32 (2H, m), 7.28-7.38 (7H, m), 7.42 (2H, t, J=7.6), 7.62-7.80 (4H, m)

Example 15: Fmoc-Ser(i-Pr)-OBzl (with sodium dihydrogenphosphate

[0414] ##STR00055##

[0415] 1) A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 1.00 g (2.0 mmol), sodium dihydrogenphosphate 1.00 g, 2-propanol 4 mL, and 2,2,2-trifluoroethanol 2 mL was stirred for 6 hours while heating at 70° C. To the reaction mixture, ethyl acetate 20 mL and 10% brine 20 mL were added, and the organic layer and the aqueous layer were separated. The organic layer was obtained as an ethyl acetate solution of Fmoc-Ser(i-Pr)-OBzl.

Fmoc-Ser(i-Pr)-OBzl

[0416] ##STR00056##

[0417] UV intensity ratio: 82.4% (detection wavelength 205 nm, retention time 3.54 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser(i-Pr)—O(i-Pr

[0418] ##STR00057##

[0419] UV intensity ratio: 1.0% (detection wavelength 205 nm, retention time 3.41 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser-OBzl

[0420] ##STR00058##

[0421] UV intensity ratio: 7.7% (detection wavelength 205 nm, retention time 2.87 min., condition 2 for high performance liquid chromatography)

[0422] The objective compound was produced in 82.4%, and Fmoc-Ser-OBzl that resulted from ring-opening of the sulfamidate without reaction with 2-propanol was produced in 7.7%. When an acid salt (in this case sodium dihydrogenphosphate) was used, the production of the byproduct was decreased by nearly 3 times compared to Example 16 in which the same starting material was used but an acid salt was not used.

[0423] 2) To an ethyl acetate solution of Fmoc-Ser(i-Pr)—OBzl, a mixture of 10% palladium on carbon 0.10 g and methanol 4 mL was added, and the reaction mixture was stirred under hydrogen gas atmosphere at room temperature for 1 hour and 30 minutes. Palladium catalyst was filtered out using Celite under reduced pressure, and the mixture obtained was concentrated under reduced pressure to give Fmoc-Ser(i-Pr)—OH 0.80 g as a crude product.

Fmoc-Ser(i-Pr)—OH

[0424] ##STR00059##

[0425] UV intensity ratio: 77.8% (detection wavelength 205 nm, retention time 2.70 min., condition 2 for high performance liquid chromatography)

[0426] 3) A solution consisting of Fmoc-Ser(i-Pr)—OH 0.80 g, water 6 mL, sodium carbonate 0.30 g (2.8 mmol), and acetonitrile 2 mL was stirred at room temperature for 3 hours, and to the reaction mixture, ethyl acetate 5 mL was added, and the organic layer and the aqueous layer were separated. The aqueous layer was obtained as an aqueous solution of Fmoc-Ser(i-Pr)—OH. To the aqueous solution of Fmoc-Ser(i-Pr)—OH obtained, 1N-hydrochloric acid 6 mL was added, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 4 hours. The precipitate was collected by filtration under reduced pressure, and the wet powder obtained was analyzed with HPLC.

Fmoc-Ser(i-Pr)—OH

[0427] ##STR00060##

[0428] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 3.25 min., condition 3 for high performance liquid chromatography)

[0429] UV intensity ratio: 87.0% (detection wavelength 205 nm, retention time 2.70 min., condition 2 for high performance liquid chromatography)

Example 16: Fmoc-Ser(i-Pr)—OBzl (without sodium dihydrogenphosphate

[0430] ##STR00061##

[0431] A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.10 mmol) and 2-propanol 0.30 mL was stirred for 2 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(i-Pr)—OBzl

[0432] ##STR00062##

[0433] UV intensity ratio: 70.1% (detection wavelength 205 nm, retention time 3.53 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser(i-Pr)—O(i-Pr

[0434] ##STR00063##

[0435] UV intensity ratio: 1.93% (detection wavelength 205 nm, retention time 3.42 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser-OBzl

[0436] ##STR00064##

[0437] UV intensity ratio: 21.6% (detection wavelength 205 nm, retention time 2.87 min., condition 2 for high performance liquid chromatography)

Example 17: Fmoc-Ser(n-Pr)—OBzl (with sodium dihydrogenphosphate

[0438] ##STR00065##

[0439] A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.10 mmol), sodium dihydrogenphosphate 24 mg, 1-propanol 0.20 mL, and 2,2,2-trifluoroethanol 0.10 mL was stirred for 2 hours while heating at 70° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(n-Pr)—OBzl

[0440] ##STR00066##

[0441] UV intensity ratio: 81.3% (detection wavelength 205 nm, retention time 3.56 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser(n-Pr)—O(n-Pr

[0442] ##STR00067##

[0443] UV intensity ratio: 1.4% (detection wavelength 205 nm, retention time 3.46 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser-OBzl

[0444] ##STR00068##

[0445] UV intensity ratio: 3.5% (detection wavelength 205 nm, retention time 2.87 min., condition 2 for high performance liquid chromatography)

[0446] The objective compound was produced in 81.3%, and a byproduct (Fmoc-Ser(n-Pr)—O(n-Pr) that resulted from the additional reaction of the objective compound with 1-propanol to cause transesterification was produced in 1.4%. When an acid salt (in this case sodium dihydrogenphosphate) was used, the yield of the objective compound was improved by nearly 10%, and the production of the byproduct was decreased by nearly 6 times compared to Example 18 in which the same starting material was used but an acid salt was not used to perform the reaction.

Example 18: Fmoc-Ser(n-Pr)—OBzl (without sodium dihydrogenphosphate

[0447] ##STR00069##

[0448] A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 0.59 g (1.23 mmol), 1-propanol 2.4 mL, and 2,2,2-trifluoroethanol 1.2 mL was stirred for 2 hours while heating at 70° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(n-Pr)—OBzl

[0449] ##STR00070##

[0450] UV intensity ratio: 68.2% (detection wavelength 205 nm, retention time 3.56 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser(n-Pr)—O(n-Pr

[0451] ##STR00071##

[0452] UV intensity ratio: 8.0% (detection wavelength 205 nm, retention time 3.46 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser-OBzl

[0453] ##STR00072##

[0454] UV intensity ratio: 4.1% (detection wavelength 205 nm, retention time 2.87 min., condition 2 for high performance liquid chromatography)

Example 19: Fmoc-Ser(2-hydroxy-2-methylpropyl)-OBzl (without sodium dihydrogenphosphate

[0455] ##STR00073##

[0456] A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 0.59 g (1.23 mmol), 2-methylpropane-1,2-diol 2.4 mL, and 2,2,2-trifluoroethanol 1.2 mL was stirred for 4 hours while heating at 70° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(2-hydroxy-2-methylpropyl)-OBzl

[0457] ##STR00074##

[0458] UV intensity ratio: 42.8% (detection wavelength 205 nm, retention time 3.09 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser(2-hydroxy-2-methylpropyl)-OH

[0459] ##STR00075##

[0460] UV intensity ratio: 11.3% (detection wavelength 205 nm, retention time 2.31 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser-OBzl

[0461] ##STR00076##

[0462] UV intensity ratio: 20.0% (detection wavelength 205 nm, retention time 2.87 min., condition 2 for high performance liquid chromatography)

Fmoc-Ser-OH

[0463] ##STR00077##

[0464] UV intensity ratio: 3.8% (detection wavelength 205 nm, retention time 2.08 min., condition 2 for high performance liquid chromatography)

Example 20: Fmoc-Ala(Cl)-OBzl

[0465] ##STR00078##

[0466] A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.1 mmol), pyridine hydrochloride 23 mg (0.2 mmol), 1-propanol 0.20 mL, and 2,2,2-trifluoroethanol 0.10 mL was stirred for 1 hour while heating at 70° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ala(Cl)—OBzl

[0467] ##STR00079##

[0468] UV intensity ratio: 91.5% (detection wavelength 205 nm, retention time 3.36 min., condition 2 for high performance liquid chromatography)

[0469] ESI (LC/MS positive mode) m/z: 436.41 (M+H.sup.+)

[0470] When pyridine hydrochloride was used as an acid salt, the starting material did not react with 1-propanol, and the chlorinated compound (Fmoc-Ala(Cl)—OBzl) that was ring-opened with chloride ion derived from pyridine hydrochloride was obtained. When sodium dihydrogenphosphate was used as an acid salt, the objective compound was found to be produced efficiently.

[0471] Production example using Fmoc-Ser-Ot-Bu as a starting material

Example 21: t-Butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-carboxylate

[0472] ##STR00080##

[0473] 1) To a solution consisting of thionyl chloride 1.61 g (13.5 mmol) and dichloromethane 20 mL cooled to −40° C., a solution consisting of Fmoc-Ser-Ot-Bu 2.00 g (5.2 mmol) and dichloromethane 5 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 15 minutes, and then pyridine 2.68 g (33.8 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 20 minutes, then at 0° C. for 20 minutes and at room temperature for 1 hour and 30 minutes. To the reaction mixture, water 30 mL was added, and the organic layer and the aqueous layer were separated. To the aqueous layer obtained, dichloromethane 20 mL was added, and the organic layer and the aqueous layer were separated again. The organic layers obtained were combined, washed with 5% NaHCO.sub.3 30 mL and 10% brine 30 mL, and then concentrated under reduced pressure to give a crude product of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamiditecarboxylate 2.22 g as a diastereomeric mixture.

[0474] 2) To a solution consisting of the crude product of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamiditecarboxylate 2.22 g and acetonitrile 30 mL cooled to 0° C., a solution consisting of sodium periodate 1.67 g (7.8 mmol), ruthenium chloride hydrate 11 mg (0.05 mmol), and water 30 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 1 hour and 30 minutes, and then at room temperature for 20 minutes. To the reaction mixture, 5% NaHCO.sub.3 60 mL, water 60 mL, and ethyl acetate 40 mL were added, and the organic layer and the aqueous layer were separated. To the aqueous layer obtained, ethyl acetate 40 mL was added, and the organic layer and the aqueous layer were separated again. The organic layers obtained were combined, washed with 10% brine 30 mL, and then concentrated under reduced pressure to give t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 2.25 g as a crude product.

[0475] 3) The crude product obtained was purified with silica gel column chromatography (elution solvent: ethyl acetate-hexane) to give t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 1.87 g (yield: 83.9% in 2 steps) as a white powder.

T-Butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate

[0476] ##STR00081##

[0477] UV intensity ratio: 96.9% (detection wavelength 205 nm, retention time 3.24 min., condition 1 for high performance liquid chromatography)

[0478] 1H-NMR (CDCl3, 400 MHz) δ: 1.50 (9H, s), 4.35 (1H, t, J=7.6), 4.48 (1H, dd, J=10.4, 7.6), 4.61 (1H, dd, J=10.4, 7.6), 4.70-4.88 (3H, m), 7.31-7.37 (2H, m), 7.39-7.45 (2H, m), 7.58-7.80 (4H, m)

[0479] ESI (LC/MS negative mode) m/z: 444.33 (M−H+)

[0480] Production example using H-MeSer-OBzl as a starting material

Example 22: Benzyl (4S)-5-methyl-1,2,5-sulfamidate-4-carboxylate

[0481] ##STR00082##

[0482] 1) To a solution consisting of thionyl chloride 2.38 g (20 mmol) and dichloromethane 50 mL cooled to −15° C., H-MeSer-OBzl hydrochloride 2.50 g (10 mmol) was added. The reaction mixture was stirred at the same temperature for 5 minutes, and then pyridine 4.75 g (60 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then at room temperature for 1 hour. To the reaction mixture, water 25 mL was added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 1N-hydrochloric acid 25 mL and 10% brine 25 mL, and then concentrated under reduced pressure to give a crude product of benzyl (4S)-5-methyl-1,2,5-sulfamiditecarboxylate 2.06 g as a diastereomeric mixture.

[0483] 2) To a solution consisting of the crude product of benzyl (4S)-5-methyl-1,2,5-sulfamiditecarboxylate 2.06 g and acetonitrile 20 mL cooled to −5° C., a solution consisting of sodium periodate 3.21 g (15 mmol), ruthenium chloride hydrate 62 mg (0.3 mmol), and water 30 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then at room temperature for 30 minutes. To the reaction mixture, water 20 mL and ethyl acetate 40 mL were added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with 10% brine 20 mL, and then concentrated under reduced pressure to give benzyl 5-methyl-1,2,5-sulfamidatecarboxylate 1.87 g as a crude product.

[0484] 3) A mixture consisting of the crude product obtained and ethyl acetate 4 mL was heated to 40° C., hexane 12 mL was added thereto, and after a precipitate appeared, the reaction mixture was stirred at room temperature for 50 minutes, and the precipitate was collected by filtration under reduced pressure. The crystals obtained were dried under reduced pressure to give benzyl (4S)-5-methyl-1,2,5-sulfamidatecarboxylate 1.14 g (yield: 42.0% in 2 steps) as pale yellow crystals.

[0485] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time 2.58 min., condition 3 for high performance liquid chromatography)

Benzyl (4S)-5-methyl-1,2,5-sulfamidatecarboxylate

[0486] ##STR00083##

[0487] UV intensity ratio: 96.3% (detection wavelength 205 nm, retention time 2.29 min., condition 2 for high performance liquid chromatography)

[0488] .sup.1H-NMR (CDCl.sub.3, 400 MHz) δ: 2.95 (3H, s), 4.10 (1H, dd, J=7.6, 6.4), 4.60-4.72 (2H, m), 5.24 (1H, d, J=12.0), 5.28 (1H, d, J=12.0), 7.32-7.44 (5H, m)

Example 23: H-MeSer(n-Pr)—OBzl

[0489] ##STR00084##

[0490] A mixture consisting of benzyl (4S)-5-methyl-1,2,5-sulfamidatecarboxylate 0.28 g (1.0 mmol) and 2-propanol 5.6 mL was stirred for 13 hours while heating at 90° C. To the reaction mixture, dichloromethane 16.4 mL and 5% aqueous sodium hydrogen carbonate 16.4 mL were added, and the organic layer and the aqueous layer were separated. The organic layer was obtained as an ethyl acetate solution of H-MeSer(n-Pr)—OBzl, and analyzed with HPLC.

[0491] UV intensity ratio: 91.8% (detection wavelength 205 nm, retention time 1.65 min., condition 1 for high performance liquid chromatography)

[0492] ESI (LC/MS positive mode) m/z: 252.48 (M+H.sup.+)

Example 24: Fmoc-Ser(CH.SUB.2.CH(OH)CF.SUB.3.)—OH (with sodium dihydrogenphosphate

[0493] ##STR00085##

[0494] 1) A mixture consisting of benzyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 100 mg (0.208 mmol), sodium dihydrogenphosphate 100 mg, 3,3,3-trifluoropropane-1,2-diol 0.543 g (4.17 mmol), and 2,2,2-trifluoroethanol 0.2 mL was stirred for 48 hours while heating at 70° C. To the reaction mixture, ethyl acetate 1 mL and 2N aqueous hydrochloric acid 1 mL were added, and the organic layer and the aqueous layer were separated. Additional ethyl acetate 1 mL was added to the aqueous layer, and the organic layer and the aqueous layer were separated. The organic layers obtained were combined to give an ethyl acetate solution of Fmoc-Ser(CH.sub.2CH(OH)CF.sub.3)—OBzl.

Fmoc-Ser(CH.SUB.2.CH(OH)CF.SUB.3.)—OBzl

[0495] ##STR00086##

[0496] UV intensity ratio: 50.9% (detection wavelength 205 nm, retention time 3.24 min., condition 2 for high performance liquid chromatography)

[0497] ESI (LC/MS positive mode) m/z: 530.59 (M+H.sup.+)

[0498] 2) To an ethyl acetate solution of Fmoc-Ser(CH.sub.2CH(OH)CF.sub.3)—OBzl, 10% palladium on carbon 20 mg was added, and the reaction mixture was stirred under hydrogen gas atmosphere at room temperature for 3 hours to give Fmoc-Ser(CH.sub.2CH(OH)CF.sub.3)—OH as an ethyl acetate solution.

Fmoc-Ser(CH.SUB.2.CH(OH)CF.SUB.3.)—OH

[0499] ##STR00087##

[0500] UV intensity ratio: 52.3% (detection wavelength 205 nm, retention time 2.52 min., condition 2 for high performance liquid chromatography)

[0501] ESI (LC/MS negative mode) m/z: 438.39 (M−H.sup.+)

Example 25: Fmoc-Ser(Bzl)-OH (with sodium dihydrogenphosphate

[0502] ##STR00088##

[0503] A mixture consisting of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.11 mmol), sodium dihydrogenphosphate 50 mg, and benzyl alcohol 0.30 mL (2.91 mmol) was stirred for 2 hours while heating at 90° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(Bzl)-OH

[0504] ##STR00089##

[0505] UV intensity ratio: 39.5% (detection wavelength 205 nm, retention time 2.93 min., condition 2 for high performance liquid chromatography)

[0506] ESI (LC/MS negative mode) m/z: 416.43 (M−H.sup.+)

Fmoc-Ser(Bzl)-Ot-Bu

[0507] ##STR00090##

[0508] UV intensity ratio: 22.9% (detection wavelength 205 nm, retention time 3.72 min., condition 2 for high performance liquid chromatography)

[0509] ESI (LC/MS positive mode) m/z: 474.68 (M+H.sup.+)

Fmoc-Ser(Bzl)-OBzl

[0510] ##STR00091##

[0511] UV intensity ratio: 22.9% (detection wavelength 205 nm, retention time 3.65 min., condition 2 for high performance liquid chromatography)

[0512] ESI (LC/MS positive mode) m/z: 474.68 (M+H.sup.+)

Example 26: Fmoc-Ser(CH.SUB.2.-3F—C.SUB.6.H.SUB.4.)—OH (with sodium dihydrogenphosphate

[0513] ##STR00092##

[0514] A mixture consisting of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 50 mg (0.11 mmol), sodium dihydrogenphosphate 50 mg, and 3-fluorobenzyl alcohol 0.30 mL (2.78 mmol) was stirred for 2 hours while heating at 90° C., and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(CH.SUB.2.-3F—C.SUB.6.H.SUB.4.)—OH

[0515] ##STR00093##

[0516] UV intensity ratio: 38.4% (detection wavelength 205 nm, retention time 2.95 min., condition 2 for high performance liquid chromatography)

[0517] ESI (LC/MS negative mode) m/z: 434.51 (M−H.sup.+)

Fmoc-Ser(CH.SUB.2.-3F—C.SUB.6.H.SUB.4.)-Ot-Bu

[0518] ##STR00094##

[0519] UV intensity ratio: 2.90% (detection wavelength 205 nm, retention time 3.71 min., condition 2 for high performance liquid chromatography)

[0520] ESI (LC/MS positive mode) m/z: 492.65 (M+H.sup.+)

Fmoc-Ser(CH.SUB.2.-3F—C.SUB.6.H.SUB.4.)—OCH.SUB.2.-3F—C.SUB.6.H.SUB.4

[0521] ##STR00095##

[0522] UV intensity ratio: 8.35% (detection wavelength 205 nm, retention time 3.71 min., condition 2 for high performance liquid chromatography)

[0523] ESI (LC/MS positive mode) m/z: 544.67 (M+H.sup.+)

Example 27: Fmoc-Ser(CH.SUB.2.-2-thienyl)-OH (with sodium dihydrogenphosphate

[0524] ##STR00096##

[0525] A mixture consisting of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 45 mg (0.11 mmol), sodium dihydrogenphosphate 45 mg, and 2-thiophene methanol 0.27 mL (2.86 mmol) was stirred at room temperature for 48 hours, and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(CH.SUB.2.-2-thienyl)-OH

[0526] ##STR00097##

[0527] UV intensity ratio: 8.61% (detection wavelength 205 nm, retention time 2.85 min., condition 2 for high performance liquid chromatography)

[0528] ESI (LC/MS positive mode) m/z: 422.37 (M+H.sup.+)

Fmoc-Ser(CH.SUB.2.-2-thienyl)-Ot-Bu

[0529] ##STR00098##

[0530] UV intensity ratio: 23.3% (detection wavelength 205 nm, retention time 3.63 min., condition 2 for high performance liquid chromatography)

[0531] ESI (LC/MS negative mode) m/z: 480.70 (M−H.sup.+)

Example 28: Fmoc-Ser(CH.SUB.2.-2-furyl)-Ot-Bu (with sodium dihydrogenphosphate

[0532] ##STR00099##

[0533] A mixture consisting of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidatecarboxylate 45 mg (0.11 mmol), sodium dihydrogenphosphate 45 mg, and 2-furfuryl alcohol 0.27 mL (3.11 mmol) was stirred at room temperature for 48 hours, and the reaction mixture was analyzed with HPLC.

Fmoc-Ser(CH.SUB.2.-2-furyl)-Ot-Bu

[0534] ##STR00100##

[0535] UV intensity ratio: 18.6% (detection wavelength 205 nm, retention time 3.49 min., condition 2 for high performance liquid chromatography)

[0536] ESI (LC/MS positive mode) m/z: 464.64 (M+H.sup.+)

[0537] Production example using Boc-homoSer-OBzl as a starting material

Example 29: Benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate

[0538] ##STR00101##

[0539] 1) To a solution consisting of thionyl chloride 7.46 g (62.7 mmol) and ethyl acetate 200 mL cooled to −15° C., a solution consisting of Boc-homoSer-OBzl 10.00 g (31.3 mmol) and ethyl acetate 30 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then pyridine 12.4 g (157 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at −15° C. for 5 minutes and then at room temperature for 3 hours. To the reaction mixture, water 100 mL was added, and the organic layer and the aqueous layer were separated. To the organic layer obtained, 1N HCl 100 mL was added, and the organic layer and the aqueous layer were separated again. The organic layer obtained was washed with 10% brine 100 mL, and then concentrated under reduced pressure to give benzyl (4S)-3-t-butoxycarbonyl-2-oxo-1,2,3-oxathiazinane-4-carboxylate 10.90 g as a crude product of a diastereomeric mixture.

[0540] 2) To a solution consisting of the crude product of benzyl (4S)-3-t-butoxycarbonyl-2-oxo-1,2,3-oxathiazinane-4-carboxylate 10.90 g and acetonitrile 90 mL cooled to −10° C., a solution consisting of sodium periodate 10.05 g (47.0 mmol), ruthenium chloride hydrate 195 mg (0.94 mmol), and water 150 mL was added dropwise over 10 minutes. The reaction mixture was stirred at −10° C. for 15 minutes, and then at room temperature for 1 hour and 40 minutes. To the reaction mixture, water 50 mL and ethyl acetate 150 mL were added, and the organic layer and the aqueous layer were separated. To the aqueous layer obtained, ethyl acetate 40 mL was added, and the organic layer and the aqueous layer were separated again. The organic layer obtained was washed with a mixture consisting of 2N aqueous HCl 20 mL and 10% brine 200 mL, and then concentrated under reduced pressure to give benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate 6.72 g as a crude product.

[0541] 3) The crude product obtained was purified with silica gel column chromatography (elution solvent: ethyl acetate-hexane) to give benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate 1.75 g (yield: 15.1% in 2 steps) as a white powder and (S)-Boc-Gly(2-chloroethyl)-OBzl 2.50 g (yield: 22.5% in 2 steps) as a pale yellow powder.

Benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate

[0542] ##STR00102##

[0543] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 2.90 min., condition 3 for high performance liquid chromatography)

[0544] UV intensity ratio: 96.3% (detection wavelength 205 nm, retention time 2.83 min., condition 2 for high performance liquid chromatography)

[0545] 1H-NMR (CDCl3, 400 MHz) δ: 1.43 (9H, s), 2.35-2.50 (1H, m), 2.60-2.70 (1H, m), 4.55-4.65 (1H, m), 4.72 (1H, dt, J=11.2, 4.8), 5.21 (2H, d, J=2.4), 5.24 (1H, dd, 5.6, 2.4), 7.30-7.40 (5H, m)

(S)-Boc-Glu(2-chloroethyl)-OBzl

[0546] ##STR00103##

[0547] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 3.07 min., condition 5 for high performance liquid chromatography)

[0548] UV intensity ratio: 98.3% (detection wavelength 205 nm, retention time 3.71 min., condition 2 for high performance liquid chromatography)

[0549] 1H-NMR (CDCl3, 400 MHz) δ: 1.44 (9H, s), 2.08-2.20 (1H, m), 2.28-2.40 (1H, m), 3.57 (2H, t, 7.2), 4.43-4.53 (1H, m), 5.08-5.20 (1H, m), 4.18 (2H, dd, J=14.8, 12.0), 7.31-7.41 (5H, m)

[0550] ESI (LC/MS positive mode) m/z: 328.57, 330.55 (M+H.sup.+)

Example 30: H-homoSer(n-Pr)—OH

[0551] ##STR00104##

[0552] A mixture consisting of benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate 50 mg (0.134 mmol) and 1-propanol 1 mL was stirred for 40 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

[0553] H-homoSer(n-Pr)—OBzl

##STR00105##

[0554] UV intensity ratio: 87.5% (detection wavelength 205 nm, retention time 1.50 min., condition 2 for high performance liquid chromatography)

[0555] ESI (LC/MS positive mode) m/z: 252.60 (M+H.sup.+)

Example 31: H-homoSer(i-Pr)—OH

[0556] ##STR00106##

[0557] A mixture consisting of benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate 50 mg (0.134 mmol) and 2-propanol 1 mL was stirred for 40 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

H-homoSer(i-Pr)—OBzl

[0558] ##STR00107##

[0559] UV intensity ratio: 84.2% (detection wavelength 205 nm, retention time 1.47 min., condition 2 for high performance liquid chromatography)

[0560] ESI (LC/MS positive mode) m/z: 252.60 (M+H.sup.+)

Example 32: H-homoSer(3-methylbutyl)-OBzl

[0561] ##STR00108##

[0562] A mixture consisting of benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate 50 mg (0.134 mmol) and 3-methylbutanol 1 mL was stirred for 40 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

H-homoSer(3-methylbutyl)-OBzl

[0563] ##STR00109##

[0564] UV intensity ratio: 84.8% (detection wavelength 205 nm, retention time 1.84 min., condition 2 for high performance liquid chromatography)

[0565] ESI (LC/MS positive mode) m/z: 280.62 (M+H.sup.+)

Example 33: H-homoSer(2-hydroxy-2-methylpropyl)-OBzl

[0566] ##STR00110##

[0567] A mixture consisting of benzyl (4S)-3-t-butoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-4-carboxylate 50 mg (0.134 mmol) and 2-methylpropane-1,2-diol 1 mL was stirred for 40 hours while heating at 80° C., and the reaction mixture was analyzed with HPLC.

H-homoSer(2-hydroxy-2-methylpropyl)-OBzl

[0568] ##STR00111##

[0569] UV intensity ratio: 70.9% (detection wavelength 205 nm, retention time 1.29 min., condition 2 for high performance liquid chromatography)

[0570] ESI (LC/MS positive mode) m/z: 282.60 (M+H+)

[0571] Production example using Fmoc-Asp(Ot-Bu)-OL as a starting material

Example 34: t-Butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-acetate

[0572] ##STR00112##

[0573] 1) To a solution consisting of thionyl chloride 2.38 g (20.0 mmol) and ethyl acetate 80 mL cooled to −40° C., a solution consisting of Fmoc-Asp(Ot-Bu)-OL 4.00 g (10.0 mmol) and dichloromethane 12 mL was added dropwise over 5 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, and then pyridine 3.96 g (50.0 mmol) was added dropwise over 5 minutes. The reaction mixture was stirred at −40° C. for 5 minutes and then at room temperature for 2 hours. To the reaction mixture, water 40 mL was added, and the organic layer and the aqueous layer were separated. To the organic layer obtained, 1N HCl 40 mL was added, and the organic layer and the aqueous layer were separated again. The organic layer obtained was washed with 10% brine 40 mL, and then concentrated under reduced pressure to give t-butyl (4R)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidite-4-acetate 4.65 g as a crude product of a diastereomeric mixture.

[0574] 2) To a solution consisting of the crude product of t-butyl (4R)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidite-4-acetate 4.65 g and acetonitrile 20 mL cooled to −10° C., a solution consisting of sodium periodate 3.21 g (15.0 mmol), ruthenium chloride hydrate 21 mg (0.1 mmol), and water 60 mL was added dropwise over 10 minutes. The reaction mixture was stirred at −20° C. for 5 minutes, and then at room temperature for 3 hours. To the reaction mixture, sodium carbonate 1.20 g (11.3 mmol), water 60 mL, and ethyl acetate 90 mL were added, and the organic layer and the aqueous layer were separated. The organic layer obtained was washed with a mixture consisting of NaCl 6.00 g (102 mmol) and water 54 mL, and then concentrated under reduced pressure to give t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-acetate 4.31 g as a crude product.

T-Butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-acetate

[0575] ##STR00113##

[0576] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 3.63 min., condition 3 for high performance liquid chromatography)

[0577] UV intensity ratio: 79.2% (detection wavelength 205 nm, retention time 3.43 min., condition 2 for high performance liquid chromatography)

[0578] 1H-NMR (CDCl3, 400 MHz) δ: 1.44 (9H, s), 2.68-2.80 (1H, m), 2.82-2.96 (1H, m), 4.33 (1H, t, J=7.2), 4.48-4.53 (4H, m), 4.81 (1H, dd, 9.6, 6.0), 7.31-7.45 (4H, m), 7.66-7.79 (4H, m)

[0579] ESI (LC/MS negative mode) m/z: 504.49 (M+HCO.sub.2.sup.−)

Example 35: (3S)-3-(9-Fluorenyl)methoxycarbonylamino-4-n-propoxybutyric acid (with sodium dihydrogenphosphate

[0580] ##STR00114##

[0581] A mixture consisting of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-acetate 50 mg (0.108 mmol), sodium dihydrogenphosphate 50 mg, and 1-propanol 1 mL was stirred for 2 hours while heating at 70° C., and the reaction mixture was analyzed with HPLC.

(3S)-3-(9-Fluorenyl)methoxycarbonylamino-4-n-propoxybutyric acid

[0582] ##STR00115##

[0583] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 3.21 min., condition 3 for high performance liquid chromatography)

[0584] UV intensity ratio: 24.6% (detection wavelength 205 nm, retention time 2.67 min., condition 2 for high performance liquid chromatography)

[0585] ESI (LC/MS negative mode) m/z: 428.42 (M+HCO.sub.2.sup.−)

T-Butyl (3S)-3-(9-fluorenyl)methoxycarbonylamino-4-n-propoxybutyrate

[0586] ##STR00116##

[0587] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 3.89 min., condition 3 for high performance liquid chromatography)

[0588] UV intensity ratio: 29.9% (detection wavelength 205 nm, retention time 3.61 min., condition 2 for high performance liquid chromatography)

[0589] ESI (LC/MS positive mode) m/z: 440.70 (M+H.sup.+)

Example 36: (3S)-3-(9-Fluorenyl)methoxycarbonylamino-4-i-propoxybutyric acid (with sodium dihydrogenphosphate

[0590] ##STR00117##

[0591] A mixture consisting of t-butyl (4S)-5-(9-fluorenyl)methoxycarbonyl-1,2,5-sulfamidate-4-acetate 30 mg (0.065 mmol), sodium dihydrogenphosphate 30 mg, and 2-propanol 0.6 mL was stirred for 2 hours while heating at 70° C., and the reaction mixture was analyzed with HPLC.

(3S)-3-(9-Fluorenyl)methoxycarbonylamino-4-i-propoxybutyric acid

[0592] ##STR00118##

[0593] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 3.16 min., condition 3 for high performance liquid chromatography)

[0594] UV intensity ratio: 7.2% (detection wavelength 205 nm, retention time 2.64 min., condition 2 for high performance liquid chromatography)

[0595] ESI (LC/MS negative mode) m/z: 382.63 (M−H.sup.+)

T-Butyl (3S)-3-(9-fluorenyl)methoxycarbonylamino-4-i-propoxybutyrate

[0596] ##STR00119##

[0597] Optical purity: 99.9% ee (detection wavelength 205 nm, retention time: 3.82 min., condition 3 for high performance liquid chromatography)

[0598] UV intensity ratio: 22.2% (detection wavelength 205 nm, retention time 3.57 min., condition 2 for high performance liquid chromatography)

[0599] ESI (LC/MS positive mode) m/z: 440.70 (M+H+)

[0600] Production example using benzyl (2S)-3-(9-fluorenyl)methoxycarbonylamino-2-hydroxymethylpropionate as a starting material

Example 37: Benzyl (5S)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate

[0601] ##STR00120##

[0602] 1) To a solution consisting of thionyl chloride 2.38 g (20.0 mmol) and ethyl acetate 80 mL cooled to −40° C., a solution consisting of benzyl (2S)-3-(9-fluorenyl)methoxycarbonylamino-2-hydroxymethylpropionate 4.32 g (10.0 mmol) and dichloromethane 12 mL is added dropwise over 5 minutes. The reaction mixture is stirred at the same temperature for 5 minutes, and then pyridine 3.96 g (50.0 mmol) is added dropwise over 5 minutes. The reaction mixture is stirred at −40° C. for 5 minutes And then at room temperature for 2 hours. To the reaction mixture, water 40 mL is added, and the organic layer and the aqueous layer are separated. To the organic layer obtained, 1N HCl 40 mL is added, and the mixture is separated into the organic layer and the aqueous layer again. The organic layer obtained is washed with 10% brine 40 mL, and concentrated under reduced pressure to give benzyl (5S)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate as a crude product of a diastereomeric mixture.

[0603] 2) To a solution consisting of the crude product of benzyl (5S)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate and acetonitrile 20 mL cooled to −10° C., a solution consisting of sodium periodate 3.21 g (15.0 mmol), ruthenium chloride hydrate 21 mg (0.1 mmol), and water 60 mL is added dropwise over 10 minutes. The reaction mixture is stirred at −20° C. for 5 minutes, and then at room temperature for 3 hours. To the reaction mixture, sodium carbonate 1.20 g (11.3 mmol), water 60 mL, and ethyl acetate 90 mL are added, and the mixture is separated into the organic layer and the aqueous layer. The organic layer obtained is washed with a mixture consisting of NaCl 6.00 g (102 mmol) and water 54 mL, and concentrated under reduced pressure to give benzyl (5S)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate as a crude product.

Example 38: Benzyl (2S)-2-(9-fluorenyl)methoxycarbonylaminomethyl-3-n-propoxypropionate (with sodium dihydrogenphosphate

[0604] ##STR00121##

[0605] A mixture consisting of benzyl (5S)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate 50 mg (0.104 mmol), sodium dihydrogenphosphate 50 mg, and 1-propanol 1 mL is stirred for 2 hours while heating at 70° C., and the reaction mixture is analyzed with HPLC.

Example 39: Benzyl (2S)-2-(9-fluorenyl)methoxycarbonylaminomethyl-3-i-propoxypropionate (with sodium dihydrogenphosphate

[0606] ##STR00122##

[0607] A mixture consisting of benzyl (5S)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate 50 mg (0.104 mmol), sodium dihydrogenphosphate 50 mg, and 2-propanol 1 mL is stirred for 2 hours while heating at 70° C., and the reaction mixture is analyzed with HPLC.

[0608] Production example using benzyl (2R)-3-(9-fluorenyl)methoxycarbonylamino-2-hydroxymethylpropionate as a starting material

Example 40: Benzyl (5R)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate

[0609] ##STR00123##

[0610] 1) To a solution consisting of thionyl chloride 2.38 g (20.0 mmol) and ethyl acetate 80 mL cooled to −40° C., a solution consisting of benzyl (2R)-3-(9-fluorenyl)methoxycarbonylamino-2-hydroxymethylpropionate 4.32 g (10.0 mmol) and dichloromethane 12 mL is added dropwise over 5 minutes. The reaction mixture is stirred at the same temperature for 5 minutes, and then pyridine 3.96 g (50.0 mmol) is added dropwise over 5 minutes. The reaction mixture is stirred at −40° C. for 5 minutes and then at room temperature for 2 hours. To the reaction mixture, water 40 mL is added, and the mixture is separated into the organic layer and the aqueous layer. To the organic layer obtained, 1N HCl 40 mL is added, and the obtained mixture is separated into the organic layer and the aqueous layer again. The organic layer obtained is washed with 10% brine 40 mL, and concentrated under reduced pressure to give benzyl (5R)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate as a crude product of a diastereomeric mixture.

[0611] 2) To a solution consisting of the crude product of benzyl (5R)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate and acetonitrile 20 mL cooled to −10° C., a solution consisting of sodium periodate 3.21 g (15.0 mmol), ruthenium chloride hydrate 21 mg (0.1 mmol), and water 60 mL is added dropwise over 10 minutes. The reaction mixture is stirred at −20° C. for 5 minutes, and then at room temperature for 3 hours. To the reaction mixture, sodium carbonate 1.20 g (11.3 mmol), water 60 mL, and ethyl acetate 90 mL are added, and the mixture is separated into the organic layer and the aqueous layer. The organic layer obtained is washed with a mixture consisting of NaCl 6.00 g (102 mmol) and water 54 mL, and concentrated under reduced pressure to give benzyl (5R)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate as a crude product.

Example 41: Benzyl (2R)-2-(9-fluorenyl)methoxycarbonylaminomethyl-3-n-propoxypropionate (with sodium dihydrogenphosphate

[0612] ##STR00124##

[0613] A mixture consisting of benzyl (5R)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate 50 mg (0.104 mmol), sodium dihydrogenphosphate 50 mg, and 1-propanol 1 mL is stirred for 2 hours while heating at 70° C., and the reaction mixture is analyzed with HPLC.

Example 42: Benzyl (2R)-2-(9-fluorenyl)methoxycarbonylaminomethyl-3-i-propoxypropionate (with sodium dihydrogenphosphate

[0614] ##STR00125##

[0615] A mixture consisting of benzyl (5R)-3-(9-fluorenyl)methoxycarbonyl-2,2-dioxo-1,2,3-oxathiazinane-5-carboxylate 50 mg (0.104 mmol), sodium dihydrogenphosphate 50 mg, and 2-propanol 1 mL is stirred for 2 hours while heating at 70° C., and the reaction mixture is analyzed with HPLC.

INDUSTRIAL APPLICABILITY

[0616] The present invention provides novel methods for producing O-substituted serine derivatives. Using the production methods of the present invention, unnatural amino acids useful for exploring peptide pharmaceuticals, and/or for supplying active ingredients of pharmaceuticals can be provided with high regioselectivity, chemical yield, and optical purity.