WO 99/37630
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`PCT/US99/01318
`
`washed with water, brine, and dried (MgSO,,). Solvent was removed under vacuum to
`
`afford the product as a colorless oil. Yield 7.60 g (99 %).
`
`‘H NMR.
`
`4-Amino-3-fluorobenzaldehyde Dimethyl Acetal
`
`2-Fluoro—4-nitrobenzaldehyde dimethyl acetal (0.59 g, 2.74 mmol) was dissolved in
`
`methanol: (20 ml), and 5% palladium on carbon (0.059 g) was added. The flask was
`
`charged with hydrogen gas, and the mixture was stirred at room temperature for 20 h. The
`
`catalyst was filtered through Celite, and solvent was removed under vacuum to afford the
`
`product. Yield 0.40 g (78 %).
`
`‘H NMR.
`
`'
`
`O—Be
`
`l-N- 3-fluoro-4- dimethox meth 1 hen 1 carbamate
`
`Benzyl chloroformate (0.34 ml, 2.38 mmol) was added dropwise with stirring to a
`
`solution of 4-amino-3-fluorobenzaldehyde dimethyl acetal (0.40 g, 2.16 mmol) and
`
`pyridine (0.26 ml, 3.24 mmol) in dichloromethane (10 ml) at 0 °C. The reaction mixture
`
`was allowed to warm to room temperature, and was washed with water, brine, and dried
`
`(MgSO4). Solvent was removed under vacuum to give the desired product as a white solid.
`
`Yield 0.56 g (81%).
`
`‘H NMR.
`
`
`
`S- R -H drox meth l-3- 4'-dimethox meth l-3'—fluoro hen 1 oxazolidine-2-one
`
`1 M Lithium bis(trimethy1sflyl)amide in tetrahydrofuran (0.86 ml, 0.941 mmol)
`
`was added dropwise with stirring at -78 °C to O-benzyl-N-[3-fluoro-4-(dimethoxymethyl)-
`
`phenyl]carbamate (0.273 g, 0.855 mmol) in tetrahydrofuran (5 ml). The mixture was
`
`stirred at -78 °C for 1 h, and then (R)-glycidyl butyrate (0.145 ml, 1.03 mmol) was added
`
`dropwise with stirring. The mixture was allowed to warm to room temperature overnight,
`
`and was then quenched with saturated aq. ammonium chloride (5 ml). The mixture was
`
`extracted with ethyl acetate, and the product was washed with water, brine, and dried
`
`(MgS0,,). Solvent was removed in vacuum, and the crude product purified by silica gel
`
`column chromatography (eluent: 30 % ethyl acetate in hexanes) to give the alcohol as an
`
`oil. Yield 0.24 g, 99%.
`
`‘H NMR.
`
`5- S -Azidometh l-3- 4'-dimethox meth l-3'-fluoro hen 1 oxazolidine-2-one
`
`
`
`
`Methanesulfonyl chloride (0.0664 ml, 0.858 mmol) was added with stirring to a
`
`10
`
`15
`
`20
`
`25
`
`solution of S—(R)-hydroxymethyl-3-[4'-dimethoxymethyl-3'-fluorophenyl]oxazolidine-2-
`
`30
`
`one (0.233 g, 0.817 mmol) and triethylarnine (0.228 ml, 1.63 mmol) in dichloromethane
`
`(10 ml) at 0 °C. The reaction was allowed to warm to room temperature, and was then
`74
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`poured into water. The organic layer was separated and washed with water, saturated aq.
`
`NaHCO3, brine, and dried (MgSO,,). Solvent was removed under vacuum to give a
`
`mesylate intermediate as an oil (0.246 g, 83%). A mixture of the mesylate (0.189 g, 0.520
`
`mmol) and sodium azide (0.170 g, 2.60 mmol) in DMF (5 ml) was heated at 75 °C for 12
`
`h. The reaction was cooled to room temperature, diluted with water (50 ml) and extracted
`
`with ethyl acetate (3 x 30 ml). The combined organic layers were washed with water,
`
`brine, and then dried (MgSO,,). Solvent was removed in vacuum, and the crude product
`
`was purified by silica gel column chromatography (eluent: 50% ethyl acetate in hexanes)
`
`to give the desired product as a colorless oil (0.154 g, 95%). MS (m/z): 311 [M + H]+.
`
`‘H
`
`10
`
`NMR.
`
`3-Fluoro-4-thiocyanoaniline .
`
`N-Bromosuccinimide (1.76 g, 9.89 mmol) and potassium thiocyanate (1.75 g, 18.0
`
`mmol) in methanol (30 ml) were stirred for 15 minutes at room temperature. The reaction
`
`mixture was cooled to 0 °C, and 3-fluoroaniline (1.00 g, 9.0 mmol) was added dropwise.
`
`15
`
`The mixture was stirred at 0 °C for 2 h. Solvent was removed under vacuum, and the
`
`residue was washed with dichloromethane. The mixture was filtered to remove
`
`succinimide by-product, and the solution was washed with water, brine, and dried
`
`(MgSO,,). Solvent was removed under vacuum to afford the desired product as a colorless
`
`oil. Yield 1.45 g (96%).
`
`‘H NMR.
`
`20
`
`O-Be
`
`1-N- 3-fluoro-4- thioc ano hen l carbamate
`
`Benzyl chloroforrnate (1.87 ml, 13.1 mmol) was added to a mixture of 3-fluoro-4-
`
`thiocyanoaniline (2.00 g, 1 1.9 mmol) and pyridine (2.12 ml, 26.2 mmol) in
`
`dichloromethane (30 ml) at 0 °C. The mixture was stirred for 30 minutes at 0 °C, allowed
`
`to warm to room temperature, and then poured into water. The organic layer was
`
`separated, washed with brine, and dried (MgSO4). Solvent was removed under vacuum.
`
`The crude product was washed with ether-hexanes and dried under vacuum to afford the
`
`desired product. Yield 3.64 g (92%); m.p. 74-75 °C.
`
`‘H NMR.
`
`O-Be
`
`1-N- 3-fluoro-4- tri hen lmeth lthio hen 1 carbamate
`
`Sodium sulfide nonahydrate (0.794 g, 3.31 mmol) in water (3 ml) was added
`
`dropwise at room temperature to a solution of O-benzyl-N-[3—fluoro-4-
`
`(thiocyano)phenyl]carbamate (1.00 g, 3.31 mmol) in ethanol (10 ml). The reaction
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`mixture was stirred at room temperature for 30 minutes, and then triphenylmethyl bromide
`
`(1.07 g, 3.31 mol) in 1,4-dioxane (5 ml) was added dropwise. The reaction was stirred
`
`overnight. Organic solvent was removed under vacuum, and the residue taken up in ethyl
`
`acetate. The solution was washed with water, brine, and dried (MgSO,,). Solvent was
`
`5
`
`removed under vacuum, and the crude product purified by silica gel column
`
`chromatography (eluent: 10% ethyl acetate in hexanes) to give the desired compound as a
`
`white solid. Yield 1.10 g, (64%); mp 152-153 °C.
`
`‘H NMR.
`
`5- R -H drox meth l-3- 4'-tri hen lmeth lthio-3'-fluoro hen loxazolidine-2-one
`
`
`
`
`
`1M Lithium bis(trimethylsilyl)amide in tetrahydrofuran (54 mL,,69.9 mmol) was
`
`10
`
`added dropwise with stirring at -78 °C to a solution of O-benzyl-N-[3-fluoro-4-
`
`(triphenylmethylthio)phenyl]carbamate (33.0 g, 63.5 mmol) in tetrahydrofuran (250 ml).
`
`The mixture was stirred at -78 °C for 1 hour, and then (R)-glycidyl butyrate (1 1.0 g, 76.2
`
`mmol) was added dropwise with stirring. The mixture was allowed to warm up to room
`
`temperature overnight, and then quenched with saturated aqueous ammonium chloride
`
`15
`
`(125 ml). The mixture was extracted with ethyl acetate, and combined organic layers
`
`washed with water, brine, and dried (MgSO,,). Solvent was removed under vacuum, and
`
`the crude product purified by silica gel column chromatography (gradient from 30% to
`
`75% of ethyl acetate in hexane) to afford the product. TLC: R, 0.2 (ethyl acetate-hexanes
`
`_ 1:1). MS 486 [M+H]*.
`
`‘H NMR.
`
`20
`
`5- S -Azidometh l-3- 4'-tri hen lmeth lthio-3'-fluoro hen loxazolidine-2-one
`
`
`
`
`
`
`Methanesulfonyl chloride (3.91 mL, 50.6 mmol) was added dropwise with stirring
`
`to a solution of 5-(R)—hydroxymethyl-3-[4’-triphenylmethylthio-3’—
`
`fluorophenyl]oxazolidine-2-one (23.4 g, 48.2 mmol) and triethylamine (10.1 n1L, 73.8
`
`mmol) in dichloromethane (200 mL) at 0 “C over ca. 10 minutes. The reaction mixture
`
`25
`
`was allowed to warm up to room temperature and then poured into water. The organic
`
`layer was separated, washed with water, saturated aq. NaHCO3, brine, and dried (MgS04).
`
`Solvent is removed under vacuum to afford the mesylate intermediate as an oil (27.2 g‘,
`
`99%). The mesylate (27.2 g, 48.2 mmol) and sodium azide (15.7 g, 241.0 mmol) in DMF
`
`(150 ml) was heated with stirring at 70°C for 12 h. The reaction ‘mixture was cooled to
`
`30
`
`room temperature, diluted with water (750 mL), and extracted with ethyl acetate.
`
`Combined organic layers were washed with water, brine, and dried (MgSO,,). Solvent was
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`removed under vacuum and the crude product purified by silica gel column
`
`chromatography (eluent: 30% ethyl acetate in hexanes) to afford the azide product as a
`
`white solid. Yield 18.1 g (73%). M.p. 77-79 °C. [a]” = -114 ° (c = 1, methanol).
`
`‘H
`
`NMR.
`
`5-Bengyloxycarbonylaminoindazole
`
`Benzyl chloroformate (9.9 ml, ca. 66 mmol) in tetrahydrofuran (66 ml) was added
`
`dropwise with stirring to 5-aminoindazole (4.44 g, 33 mmol) in tetrahydrofuran (150 ml)
`
`and pyridine (12.0 ml, 150 mol) at -5 °C. The mixture was allowed to warm to room
`
`temperature, stirred for 4 h, and concentrated under vacuum. Ethyl acetate (100 ml) and
`
`water (150 ml) were added, and the aqueous layer was extracted with ethyl acetate (2 x 100
`
`ml). The combined organic laters were washed with 0.3 N aq. HCI (2 x 100 ml), water,
`
`brine, and dried (MgSO4). Solvent was removed under vacuum to afford the crude product
`
`as a mixture of two regioisomers. MS (m/z): 402.1 [M+H]*. 0.3 M lithium hydroxide
`
`monohydrate in methanol (250 ml, ca. 75 mmol) was added. The mixture was stirred at
`
`room temperature for 45 min and then carefully acidified with 6 N aq. HCI until the pH of
`
`the solution was 2. The resulting product was filtered off, washed with water and dried
`
`under vacuum to afford the desired compound. R, 4.2 min. MS (m/z): 268.1 [M+H]*.
`
`‘H
`
`NMR.
`
`5-Bengyloxycarbonylamino-1-triphenylmethylindazole
`
`5-Benzyloxycarbonylaminoindazole (0.534 g, 2 mmol) was stirred with trityl
`
`chloride (0.556 g, 2 mmol) and tetrabutylammonium iodide (0.074 g, 0.2 mmol) in
`
`tetrahydrofuran (5 ml) and triethylamine (0.42 ml, 3 mmol) for 3 days at room
`
`temperature. Solvent was removed under vacuum, and the solid residue was triturated with
`
`methanol (3 ml). The solid was washed with a mixture of methanol-water (5:1, ca. 15 ml)
`
`and dried under vacuum to afford the desired product. Yield 0.73 g (72%).
`
`‘H NMR.
`
`5-| 5-g jR_Q-Hydroxymethyloxazolidine-2-one-3-yl |-l-triphenylmethylindazole
`
`1 M Lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.1 mL, 1.1 mmol) was
`
`added dropwise with stirring at -78 °C to 5-benzyloxycarbonylamino-l-tritylindazole
`
`(0.510 g, 1 mmol) in tetrahydrofuran (10 mL) under nitrogen atmosphere. The mixture
`
`was stirred at -78 °C for 1.5 h. (R)-Glycidyl butyrate (0.160 mL, 1.2 mmol) was added
`
`dropwise with stirring. The mixture was allowed to warm to r.t. overnight. Saturated aq.
`77
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`WO 99/37630
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`NH,,Cl (10 mL) was added, and the mixture was extracted with EtOAc (2 x 20 mL). The
`
`combined organic layers were washed with water ( 10 mL), brine (10 mL), and dried
`
`(MgSO4). Solvent was evaporated to 3 mL, and the residue was triturated with hexanes
`
`(50 mL). White crystalline product was filtered off, washed with hexanes, and dried in
`
`vacuo. Yield 0.440 g (93%). MS (m/z): 232.1 [M-Trt]‘.
`
`‘H NMR.
`
`5- 5- S -Azidometh loxazolidine-2-one-3- l -1-tri hen lmeth lindazole
`
`
`
`
`
`Methanesulfonyl chloride (0.066 ml, 0.85 mmol) was added dropwise with stirring
`
`to a solution of 5-[5-(R)-hydroxymethyloxazolidine-2-one-3-yl]-l-triphenylmethylindazole
`(0.300 g, 0.63 mmol) and triethylamine (0.18 ml, 1.3 mmol) in dichloromethane (7.0 ml) at
`
`- 30 °C over 5 minutes. The reaction mixture was stirred at 5 °C for 2 h and quenched
`
`with water (15 ml). Ethyl acetate (20 ml) was added, and the organic layer was washed
`
`with water, brine, and dried (MgSO4). Solvent was removed under vacuum to afford a
`
`mesylate intermediate. The mesylate and sodium azide (0.205 g, 3.15 mmol) in DMF (4
`
`ml) was heated with stirring at 75 °C for 4 h. The reaction mixture was cooled to room
`
`temperature, diluted with water (ca. 10 ml), and extracted with ethyl acetate (2 x 15 ml).
`
`The combined organic layers were washed with water, brine, and dried (MgSO,,). Solvent
`
`was removed under vacuum to afford the desired product as off—white crystals. Yield 0.31
`
`g (95%). MS (m/z): 257.1 [M-Trt]'.
`
`'H NMR.
`
`BAL Aldehyde Resin
`
`4-(4-Formyl-3,5-dimethoxyphenoxy)butyric acid (9.33 g, 34.8 mmol), pyridine (15
`
`ml), and diisopropylcarbodiimide (3.00 ml, 19.1 mmol) in dichloromethane (135 ml) were
`
`stirred at room temperature for 1 h. Tentagel S-NH, resin (Rapp Polymere, 0.29 mmol/g,
`
`8.7 mmol) was added, and the mixture was agitated at room temperature overnight. The
`
`resin was filtered, washed liberally with MeOH and dichloromethane and dried under
`
`10
`
`15
`
`20
`
`25
`
`vacuum.
`
`5- 5- S
`
`
`-Acetamidometh loxazolidine-2-one-3- l-1-indazole
`
`
`
`
`Tetrahydrofuran (1.0 mL) was added to the mixture of 5—[5-(S)-
`
`azidomethyloxazolidine-2-one-3-yl]-1-triphenylmethylindazole (0.065 g, 0.13 mmol, ca. 3
`
`eq. with respect to the resin reagent), triphenylphosphine (0.034 g, 0.13 mmol), and BAL
`
`30
`
`aldehyde resin (150 mg, ca. 0.044 mmol). The mixture was stirred at r.t. for 2 h. A rubber
`
`septum was replaced with a teflon-coated cap, and the mixture was agitated at 75 °C for ca.
`78
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`WO 99/37630
`
`PCT/U S99/0] 318
`
`10 h. A tetrahydrofuran-triethylorthofonnate mixture (1 :1, 1 mL) was added to the
`
`resulting imine resin, followediby 0.5 M NaBH,CN (0.5 mL, 0.25 mmol). The mixture
`
`was agitated at room temperature for 3 h. The resulting amine resin was washed liberally
`
`with MeOH and dichloromethane, and dried under vacuum. An acetic anhydride-pyridine-
`
`5
`
`dichloromethane solution (1 to 1.5 to 3, 4 mL) was added, and the mixture was agitated for
`
`2 h (until negative ninhydrine test indicated completion of the acylation). The trityl
`
`protection was removed by treatment with 1% TFA in DCM (2 x 4 mL, 15 min), and the
`
`product was cleaved with 60% TFA in DCM (2 mL) over 2 h. HPLC purity for the
`
`cleaved product was 90% (Rt 2.95 min). Solvent was removed under vacuum, and the
`
`10
`
`product was purified by preparative silica gel TLC (eluent: dichloromethane-MeOH 5:1).
`
`Yield 7.0 mg (58%). R, 2.9 min. (given below). MS (m/z): 275.1 [M+H]‘.
`
`‘H NMR.
`
`BAL Resin Immobilized 5- S -Aminometh l-3- 4'-dimethox meth 1-3’-
`
`
`
`
`fluorophenyl |-oxazolidine-2-one
`
`I
`
`Triphenylphosphine (0.130 g, 0.496 mmol) was added to a mixture of BAL
`
`15
`
`aldehyde resin (0.57 g, 0.165 mmol) and 5-(S)-azidomethyl-3-[4'-dimethoxymethyl-3'-
`
`fluorophenyl]-oxazolidine-2-one (0.154 g, 0.496 mmol) in THF (3 ml) at room
`
`temperature. The mixture was stirred at room temperature for 2 h, and then at 75°C for 16
`
`h. The mixture was cooled to room temperature, and 1M sodium cyanoborohydride in THF
`
`(0.99 ml, 0.992 mmol) was added in one portion. The reaction mixture was agitated for 8
`
`20
`
`h. The resulting amine resin was washed liberally with methanol and dichloromethane and
`
`dried under vacuum.
`
`-Acetamidometh l-3- 4'-dimethox meth 1-3’-
`BAL Resin Immobilized 5- S
`
`
`
`
`
`fluorophenyl |-oxazolidine-2-one
`
`Acetic anhydride-pyridine-dichloromethane solution (1 to 1.5 to 3, 4 mL) was
`
`25
`
`added to BAL resin immobilized 5-(S)-aminomethyl-3-[4'-dimethoxymethyl-3'-
`
`fluorophenyl]—oxazolidine-2-one, and the mixture was agitated for ca. 2 h (until negative
`
`ninhydrine test indicated completion of the acylation). The resin was filtered, washed
`
`liberally with methanol and dichloromethane and dried under vacuum.
`
`-Acetamidometh l-3- 4'-forrn 1-3'-fluoro hen l oxazolidine-2-one
`
`
`
`30
`
`BAL resin immobilized 5-(S)—acetarnidomethyl-3-[4'-dimethoxymethyl~3'-
`
`fluorophenyl]-oxazolidine-2-one (0.100 g, 0.029 mmol) was suspended in 60%
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`WO 99/37630
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`trifluoroacetic acid in dichloromethane (2 ml) for 2 h at room temperature. The mixture
`
`was filtered, and supernatant was evaporated under vacuum to give the crude product. The
`
`crude product was purified by preparative HPLC to afford the desired product as an oil.
`
`Yield 4.9 mg, (60%). R, 3.0 min. MS (rn/z): 281.1 [M + H]*.
`
`'H NMR.
`
`BAL Resin Immobilized 5- S -Aminometh 1-3- 4'-tert-butox carbon 1-3’-
`
`
`
`fluorophenyl |-oxazolidine-2-one
`
`Triphenylphosphine (7.61 g, 29.0 mmol) was added to a mixture of BAL aldehyde
`
`resin (33.3 g, 9.67 mmol) and 5-(S)-azidomethy1-3-[4'-tert-butoxycarbonyl-3'-I
`
`fluorophenyl]-oxazolidine-2-one (9.76 g, 29.0 mmol) in tetrahydrofuran (170 ml) under
`
`nitrogen at room temperature. The mixture was agitated at room temperature for 2 h and
`
`then at 75 °C for 16 h. The mixture was cooled to room temperature, and 1 M sodium
`cyanoborohydride in THF (58.0 ml, 58.0 mmol) was added in one portion. The reaction
`
`mixture was agitated for 8 h. The resulting amine resin was filtered, washed liberally with
`
`methanol and dichloromethane, and dried under vacuum.
`
`-3--
`BAL Resin Immobilized 5- S -Aminometh l-3- 4'—carbox
`
`
`
`
`fluorophenyl loxazolidine-2-one
`
`A mixture of 1 M chlorotrirnethylsilane in dichloromethane (290 ml, 0.29 mol) and
`
`1M phenol in dichloromethane (290 ml, 0.29 mol) was added to BAL resin immobilized 5-
`
`(S)—aminomethyl-3-[4'—tert-butoxycarbonyl-3'-fluorophenyl]-oxazolidine-2-one, and the
`
`reaction mixture was agitated at room temperature for 36 h. The resulting acid resin was
`
`filtered, washed liberally with methanol and dichloromethane, and dried under vacuum.
`
`General Procedure for the Synthesis of Immobilized 5-g S)-Acylarninomethyl-3-|4¥
`
`carboxy-3 '—fluorophenyl |-oxazolidine-2-ones
`
`A selected carboxylic acid (18.0 mmol), pyridine (1.46 ml,18.0 n-tmol) and
`
`diisopropylcarbodiimide (1.35 ml, 9.90 mmol) in a mixture of dimethylformamide—
`
`dichloromethane (431, 8 ml) were stirred at room temperature for 1 h. An appropriate BAL
`
`resin immobilized 5—(S)-aminomethyl-3-[4'-carboxy-3'-fluorophenyl]oxazolidine-2-one
`
`(1.80 mmol) was added and the mixture was agitated at room temperature for 16 h (or until
`
`ninhydrine test indicated a completion of the acylation). The resin was filtered, washed
`
`liberally with dimethylformamide, MeOH, dichloromethane, and dried under vacuum.
`
`80
`
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`WO 99/37630
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`
`5- S
`-Acetamidometh [-3- 4'-carbox -3'-fluoro hen ll-oxazolidine-2-one
`
`
`
`
`
`
`Acetic anhydride-pyridine-dichloromethane solution (1 :l.5:3, 200 mL). was added
`
`to an immobilized 5—(S)—acylaminomethyl-3-[4'-carboxy-3'-fluorophenyl]—oxazolidine-2-
`
`one (33.3 g, 9.67 mmol), and the mixture was agitated overnight. The resin was filtered,
`
`washed liberally with methanol and dichloromethane and dried under vacuum. The
`
`acylated resin (0.100 g, 0.029 mmol) was suspended in 60% trifluoroacetic acid in
`
`dichloromethane for 2 h at room temperature. The mixture was filtered, and the
`
`supernatant was evaporated under vacuum to give a white solid which was washed with
`
`ether and dried under vacuum. Yield 7.6 mg (88%); mp 252-253 °C.
`
`‘H NMR.
`
`BAL Resin Immobilized 5-(S )-Acetamidomethyl-3-[4'—
`
`gpentafluorophenyl )oxycarbonyl-3'-fluorophenyl |-oxazolidine-2-one
`
`Pentafluorophenyl trifluoroacetate (7.10 ml, 41.3 mmol) was added to a mixture of
`
`BAL resin immobilized 5-(S)-acetarnidomethyl—3-[4'-carboxy-3'-fluorophenyl]oxazolidine-
`
`2-one (20.4 g, 5.90 mmol) and pyridine (8 ml) in N—methylpyrrolidine-2-one (35 ml). The
`
`reaction mixture was agitated at room temperature for 16 h. The resin was filtered, washed
`
`with N—methylpyrrolidine-2-one and dichloromethane, and dried under vacuum. The resin
`
`was analyzed by cleavage with 60% trifluoroacetic acid in dichloromethane (2 ml per
`
`0.100 g. 0.029 mmol of the resin, 2 h). The resulting supernatant was evaporated under
`
`vacuum to give the released pentafluorophenyl ester as a white solid. The solid was
`
`purified by preparative TLC (eluent 10% MEOH in dichloromethane). Yield 8.0 mg
`
`(60%); m.p. 172-173 °C.
`
`‘H NMR.
`
`
`4’ ’-mo holino hen lamino carbon 1-3’-
`
`fluorophenyl |-oxazolidine-2-one
`
`BAL resin immobilized 5-(S)-acetamidomethyl-3-[4'-
`
`(pentafluorophenyl)oxycarbonyl-3'-fluorophenyl]oxazolidine-2-one (O.l00g, 0.029 mmol)
`
`was agitated with 4-morpholinoaniline (0.155 mg, 0.87 mmol) in 10% pyridine in
`
`dimethylformamide (2 ml) for 24 h. The resin was filtered and washed liberally with
`
`dimethylformamide, MeOH, DCM, and dried under vacuum. The dry resin was cleaved in
`
`60% trifluoroacetic acid in dichloromethane (2 ml) for 2 h at room temperature. The
`
`supernatant was evaporated under vacuum, and the crude product was purified by
`
`81
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`WO 99/37630
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`
`preparative TLC (eluent: 10% methanol in dichloromethane to give product as a white
`
`solid. Yield 6.6 mg (50%). MS (m/z): 457.2 [M+H]’.
`
`‘H NMR.
`
`5- S -Acetamidometh l-3- 4’-
`
`
`rid larnino carbon l-3’-fluoro hen l -
`
`
`
`
`
`
`oxazolidine-2-one
`
`BAL resin immobilized 5-(S)-acetamidomethyl-3-[4'-
`
`(pentafluorophenyl)oxycarbonyl-3'-fluorophenyl]oxazolidine-2—one (0.l00g, 0.029 mmol)
`
`was agitated with 3—arninopyridine (0.082 mg, 0.87 mmol) in 10% pyridine in
`
`dimethylformarnide (2 ml) for 24 h. The resin was filtered and washed liberally with
`
`dimethylformarnide, MeOH, DCM, and dried under vacuum. The dry resin was cleaved in
`
`60% trifluoroacetic acid in dichloromethane (2 ml) for 2 h at room temperature. The
`
`supernatant was evaporated under vacuum, and the crude product was purified by
`
`preparative TLC (eluent: 10% methanol in dichloromethane) to give the product as a white
`
`solid. Yield 4.3 mg (40%). MS(rn/z): 373.1 [M+H]*.
`
`‘H NMR.
`
`5- S -Acetamidometh l-3- 4’-
`
`
`
`4”—mo holino carbon 1-3’-
`
`
`
`
`fluorophenyl [oxazolidine—2-one
`
`BAL resin immobilized 5-(S)-acetamidomethyl-3-[4‘-
`
`(pentafluoropheny1)oxycarbonyl-3'-fluorophenyl]oxazolidine-2-one (0.l00g, 0.029 mmol)
`
`was agitated with morpholine (0.10 ml, 0.116 mmol) in N-methylpyrrolidine—2-one (2 ml)
`
`for 16 h. The resin was filtered and washed liberally with N—methylpyrrolidine—2-one,
`
`MeOH, dichloromethane, and dried under vacuum. The dry resin was cleaved in 60%
`
`trifluoroacetic acid in dichloromethane (2 ml) for 2 h at room temperature. The resin was
`
`filtered, the filtrate evaporated under vacuum, and the crude product was purified by
`
`preparative TLC (eluent: 10% MeOH in dichloromethane) to give the product as a white
`
`solid. Yield 5.6 mg (53%); m.p. 210-211 °C.
`
`‘H NMR.
`
`BAL Resin Immobilized Weinreb Amide: 5- S -Acetamidometh 1-3- 4’-N-
`
`
`
`
`
`methoxy-N-methylaminocarbonyb3 ’ -fluorophenyl |-oxazolidine-2-one
`
`BAL resin immobilized 5-(S)-acetamidomethyl-3-[4’-
`
`10
`
`15
`
`20
`
`25
`
`(pentafluoropheny1)oxycarbonyl-3'—fluorophenyl]oxazolidine-2-one (1.00 g, 0.29 mmol)
`
`was agitated with N-methoxy-N-methylamine hydrochloride (0.59 g, 6.0 mmol) and
`
`30
`
`triethyularnine (0.84 ml, 6.0 mmol) in N-methylpyrrolidine—2-one for 16 h at room
`
`temperature. The resin was filtered, washed liberally with N-methylpyrrolidine—2-one,
`82.
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`MeOl-l, dichloromethane, and dried under vacuum. A small portion of the resin (ca 10
`
`mg) was cleaved in 60% trifluoroacetic acid in dichloromethane (0.20 ml) for 2 h at room
`
`temperature. The supernatant was concentrated under vacuum to afford the cleaved
`
`Weinreb amide as an oil. R, 2.8 min. MS (rn/z): 340.1 [M + H]".
`
`‘H NMR.
`
`BAL Resin Immobilized Aldehyde 5-(S )—Acetamidomethyl-3-|4’-formyl-33
`
`fluorophenyl |-oxazolidine-2-one
`
`_
`
`0.1 M Lithium aluminum hydride in tetrahydrofuran (0.52 ml) was added dropwise
`
`with stirring to BAL resin immobilized 5-(S)-acetamidomethyl-3-[4’-N-methoxy-N-
`
`methylaminocarbonyl-3’-fluorophenyl]oxazo1idine-2—one (0.150 g, 0.044 mmol) in
`
`tetrahydrofuran (2 ml) at -78 °C. The mixture was agitated at -78 °C for 4-6 h. It was then
`
`allowed to warm to room temperature overnight. The resin was filtered, washed liberally
`
`with tetrahydrofuran, MeOH, dichloromethane, and dried under vacuum.
`
`‘H NMR.
`
`-oxazolidine-2-one
`5- S -Acetarnidometh l-3- 4’-form 1-3’-fluoro hen l
`
`
`
`
`
`BAL resin immobilized 5-(S)-acetarnidomethyl-3-[4’-formyl-3’-fluorophenyl]—
`
`oxazolidine-2-one (0.150 g, 0.0435 mol) was cleaved with 60% trifluoroacetic acid in
`
`dichloromethane (2 ml) for 2 h at room temperature. Supernatant was evaporated under
`
`vacuum to give the crude product as an oil. MS (m/z): 281.1 [M + H]+.
`
`‘H NMR.
`
`
`
`-Acetarnidometh l-3- 4’-acetvl-3’—fluoro hen l oxazolidine-2-one
`
`
`
`3.0 M Methylmagnesium iodide in diethyl ether (0.022 ml, 0.066 mmol) is added
`
`dropwise with stirring to BAL resin immobilized 5-(S)—acetamidomethyl-3-[4’-N-
`
`methoxy-N-methylaminocarbonyl-3’-fluorophenyl]-oxazolidine-2-one (0.150 g, 0.044
`
`mmol) in tetrahydrofuran (2 ml) at -78 °C. The mixture is agitated at 78 °C for 5-10 h, and
`
`then allowed to warm to room temperature overnight. The resin is filtered, washed
`
`liberally with tetrahydrofuran, MeOH, dichloromethane, and dried under vacuum. The
`
`resulting ketone resin is cleaved with 60% trifluoroacetic acid in dichloromethane (2 ml)
`
`"for 2 h at room temperature. The supernatant is evaporated under vacuum to afford the
`
`desired product.
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`BAL Resin Immobilized Ac l Azide 5- S
`
`
`
`-Acetamidometh l-3- 4’-azidocarbon l-
`
`
`3 ’-fluorophenyl |oxazolidine-2-one
`
`Method A: with azidotrimethylsilane and tetrabutylammonium fluoride. 1 M
`
`Tetrabutylarnrnonium fluoride in tetrahydrofuran (0.609 ml, 0.609 mmol) was added to
`
`azidotrimethylsilane (0.34 ml, 2.6 mmol) in tetrahydrofuran (3.5 ml), and the mixture was
`
`kept at room temperature for 0.5 h. The resulting solution was added to BAL resin
`
`immobilized 5—(S)-acetamidomethyl-3-[4'-(pentafluorophenyl)oxycarbonyl-3'-
`
`fluorophenyl]-oxazolidine-2-one, and the mixture was agitated at room temperature for 4-5
`h. The acyl azide resin was filtered, washed with dichloromethane and acetone. IR (cm-'):
`
`2136 (N3). The resin was further analyzed by cleavage with 60% trifluoroacetic acid in
`
`dichloromethane (2 ml per 0.100 g, 0.029 mmol of the resin, 2 h). The resulting
`
`supernatant was evaporated under vacuum to give the released acyl azide product. R, 3.3
`
`min. IR (cm”): 2138 (N3). MS (rn/z): 278.1 [M-N2+H]“.
`
`‘H NMR.
`
`Method B: with tetrabutylammonium azide. BAL resin immobilized 5-(S)-
`
`acetamidomethyl-3-[4'-(pentafluorophenyl)oxycarbonyl-3’-fluorophenyl]oxazolidine—2-one
`(Tentagel HL NH, resin, 1.00 g, ca. 0.40 mmol/g) was agitated with tetrabutylammonium
`
`azide (0.797 g, 2.8 mmol) in tetrahydrofuran (10 ml) for 5 h at room temperature. The
`resin was filtered, washed liberally with dichloromethane and acetone, and dried under
`vacuum.
`
`4
`
`BAL Resin Immobilized Protected Amine 5-g S}-Acetamidomethyl-3-|4’—g 9”-
`
`fluorenylmethoxycarbonyl )amino—3 ’ —fluorophenyl ]-oxazolidine-2-one
`
`BAL resin immobilized acyl azide 5-(S)-acetamidomethyl-3-[4’-azidocarbonyl-3’-
`
`fluorophenyl]oxazolidine-2-one (0.75 g, 0.22 mmol) and (9-fluorenyl)methanol (1.18 g,
`
`6.0 mmol) in tetrahydrofuran (7.0 ml) were agitated at 80 °C for 4 h. The resulting Fmoc-
`
`protected amine resin was washed with tetrahydrofuran, MeOH, dichloromethane, and
`
`dried under vacuum.
`
`5- S -Acetamidometh l-3- 4’-
`
`
`
`
`
`
`9”-fluoren lmethox carbon lamino-3’-fluoro-
`
`
`
`phenyl |-oxazolidine-2-one
`
`10
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`
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`
`25
`
`Method A. BAL resin immobilized 5—(S)-acetamidomethyl-3-[4’-(9"-
`
`30
`
`fluorenylmethoxycarbonyl)amino-3’-fluorophenyl]oxazolidine-2-one (0.200 g) was
`
`cleaved with 60% trifluoroacetic acid in dichloromethane (2 ml) for 2 h. The resulting
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`WO 99/37630
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`supernatant was evaporated under vacuum to give the released Fmoc carbamate product.
`
`R, 4.3 min. MS (rn/z): 490.2 [M +H]“.
`
`‘H NMR.
`
`Method B. 9-Fluorenylmethyl chlorofonnate (0-039 g, 0.15 mmol) in
`
`dichloromethane (0.300 ml) and pyridine (0.05 ml, 0.62 mmol) was added to BAL resin
`
`immobilized 5-(S)-acetamidomethyl-3-[4’-amino-3’-fluorophenyl]oxazolidine-2-one, and
`
`the mixture was agitated at room temperature for 2 h. The resulting resin was worked up
`
`and cleaved as described above for Method A. K 4.3 min. MS (rn/z): 490.2 [M +H]".
`
`‘H
`
`NMR.
`
`BAL Resin Immobilized Amine 5- S)-Acetamidometh l-3- 4’
`
`
`
`
`—arnino-3 ’ -
`
`
`
`fluorophenyl |-oxazolidine-2-one
`
`Method A. BAL resin immobilized 5-(S)-acetamidomethyl-3-[4’-(9"-
`
`fluorenylmethoxycarbonyl)amino-3’-fluorophenyl]oxazo1idine-2—one (ca. 0.200 g) was
`
`deprotected with 20% piperidine in dimethylfonnamide (2 ml) for 20 min. The resulting
`
`amine resin was washed liberally with MeOH, dichloromethane, and dried under vacuum.
`
`The resin was analyzed by cleavage with 60% trifluoroacetic acid in dichloromethane (2
`
`ml, 2 h). The resulting supernatant was evaporated under vacuum to give the released‘
`
`amine product. MS (m/z): 268.1 [M+H]+.
`
`‘H NMR.
`
`Method B. BAL resin immobilized 5-(S)-acetamidomethyl-3-[45
`
`(pentafluorophenyl)-oxycarbonyl-3'-fluorophenyl]oxazolidine-2-one (0.200 mg),
`
`azidotrimethylsilane (0.240 ml, 1.74 mmol) and catalytic tetrabutylarnmonium fluoride
`
`(0.05 ml, 0.05 mmol) in tetrahydrofuran (5 ml) were agitated at 80 °C for 4 h. The
`
`resulting amine resin was washed liberally with MeOH and dichloromethane. It was dried
`
`under vacuum and analyzed as described above for Method A. MS (rn/z): 268.1 [M+H]*.
`
`‘H NMR.
`
`—Aceta.midometh 1-3- 4’- ara-nitrobenzene sulfonamido-3’-fluoro hen 1 -
`
`
`
`
`oxazolidine-2-one
`
`BAL resin immobilized 5-(S)-acetamidomethyl-3-[4’-amino—3’-fluorophenyl]-
`
`oxazolidine-2-one (0.200 g) was agitated with paramitrobenzenesulfonyl chloride (0.108 g,
`
`0.50 mmol) in dichloromethane (2.0 ml) with N-methylmorpholine (0.200 ml) for 14 h at
`
`10
`
`15
`
`20
`
`25
`
`30
`
`room temperature. The resulting sulfonamide resin was filtered, washed liberally with
`
`dimethylforrnamide, MeOH, dichloromethane, and dried under vacuum. Theldry resin was
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`WO 99/37630
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`cleaved with 60% trifluoroacetic acid in dichloromethane (2 ml, 2 h). The resulting
`
`supernatant was evaporated under vacuum to give the sulfonarnide product. MS (rn/z):
`
`453.1 [M+H]*.
`
`‘H NMR.
`
`-acetamidometh loxazolidine-2-
`N‘- 9~Fluoren lmethox carbon 1 -N2-
`
`
`
`
`one-3 ’ ’ -yl )-3'-fluorophenyl |thiourea
`
`BAL resin immobilized 5-(S)-acetarnidomethyl-3-[4’-amino-3’-fluorophenyl]-
`
`oxazolidine-2—one (0.200 g) was agitated with 9-fluorenylmethoxycarbonylisocyanate
`
`(0.140 g, 0.50 mmol) in dich1oro_rnethane(2.0 ml) for 14 h at room temperature. The 5
`
`resulting thiourea resin was filtered, washed liberally with dimethylformarnide, MeOH,
`
`dichloromethane, and dried under vacuum. The dry resin was cleaved with 60%
`
`trifluoroacetic acid in dichloromethane (2 ml, 2 h). The resulting supernatant was
`
`evaporated under vacuum to give the sulfonamide product. R, 4.5 min. MS (n1/z): 549.1
`
`[M+H]*.
`
`‘H NMR.
`
`
`S -Acetamidometh 13- 4’- 4”- hen lthiazole-2”
`
`l amino-3’-fluoro-
`
`phenyl loxazolidine-2-one
`
`BAL resin immobilized N‘-(9-Fluorenylmethoxycarbonyl)-N2-[4’-(5’ ’-(S)-
`
`acetamidomethyloxazolidine-2-one-3"-yl)-3'-fluorophenyl]thiourea was deprotected with
`
`20% piperidine in dirnethylformamide (2 ml) for 40 min, filtered, washed liberally with
`
`MeOH, dichloromethane, and dried under vacuum. 2-Bromoacetophenone (0.100 g, 0.50
`
`mnol) in tetrahydrofuran (2.0 ml) was added, and the mixture was agitated at room
`
`temperature for 2 h. The resulting thiazole resin was washed liberally with MeOH,
`
`dichloromethane, and dried under vacuum. The dry resin was cleaved with 60%
`
`trifluoroacetic acid in dichloromethane (2 ml, 2 h). The resulting supernatant was
`
`evaporated under vacuum to give the thiazole product. R, 3.9 min. MS (rn/z): 427.1
`
`10
`
`15
`
`20
`
`25
`
`[M+H]*.
`
`‘H NMR.
`
`5-
`5”-amino-4”—c anooxazole-2”-
`-3’-
`
`
`fluorophenyl |-oxazolidine-2-one
`
`BAL resin immobilized 5-(S)-acetamidomethyl-3-[4‘-(pentafluorophenyl)-
`
`oxycarbonyl-3'-fluorophenyl]oxazolidine-2-one (0.100 g) was agitated with
`
`30
`
`aminomalonitrile tosylate (0.253 g, 1 mmol) in a mixture of dry pyridine and N-
`
`methylpyrrolidine-2-one (1:1, 2.0 ml) at 60 °C for 8-10 h. The resulting aminooxazole
`86
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`wo 99/37630
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`
`resin was washed liberally with MeOH, dichloromethane, and dried under vacuum. The
`
`dry resin was cleaved with 60% trifluoroacetic acid in dichloromethane (2 ml, 2 h). The
`
`resulting supernatant was evaporated under vacuum to give the oxazole product. R, 3.2
`
`min. MS (rn/2): 360.1 [M+H]*.
`
`‘H NMR.
`
`5- S -Acetamidometh l-3- 4’-tri hen lmeth lthio-3’—fluoro hen 1 oxazolidine-2-
`
`
`
`
`
`BE
`
`Triphenylphosphine (2.82 g, 10.8 mmol) was added portionwise to a solution of 5-
`
`(S)—azidomethyl-3-[4’-tripheny1methylthio—3’-fluorophenyl]- oxazolidine-2—one (5.00 g,
`
`9.79 mmol) in THF (40 mL), and the mixture stirred for 2 h at room temperature. Water
`
`(1.41 mL, 78.3 mmol) was added, and the mixture heated at 40 °C overnight. Solvent was
`
`removed under vacuum, and the oily residue dissolved in dichloromethane (50 mL).
`
`Acetic anhydride (4.62 ml, 49.0 mmol) and pyridine (7.92 ml, 97.9 mmol) were added, and
`
`the mixture stirred for 8 h at r.t. Solvent was removed under vacuum and the crude
`
`product purified by silica gel flash column chromatography (cluent: 30% ethyl acetate in
`
`hexanes) to give the product as a foam (4.98 g, 97 %); MS: 527 [M+H]".
`
`‘H NMR.
`
`BAL Resin Immobilized 5- S -Aminomethvl-3- 4’-tri hen lmeth lthio-3’-
`
`
`
`fluorophenyl |-oxazolidine-2-one
`
`Diisopropylcarbodiiimide (4.24 ml, 27.0 mmol) aws added to 4-(4-forrnyl-3,5-
`
`dimethoxyphenoxy)butyric acid (13.19 g, 49.2 mmol) and pyridine (20 mL) in
`
`dichloromethane (190 mL), a