4-(2-aminoethyl)phenol;hydrochloride

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To an ice cold suspension of 3.69 g (0.03 mol) nicotinic acid in a solution of 5.2 g (0.03 mol) tyramine hydrochloride in 100 ml of pyridine, 6.18 g (0.03 mol) of dicyclohexylcarbodiimide (DCC) were gradually added while stirring. Stirring was continued at room temperature for 24 hrs and the formed dicyclohexylurea was removed by filtration. The pyridine was removed by distillation in vacuo and the residue was triturated with cold water, filtered and crystallized from 50% aqueous methanol. Yield, 6.25 g (86%), m.p. 179°-181° C. PMR (DMSO-d6 /D2O) δ 9.0-8.66 (m, 2H, C2 and C6 pyridine protons), 8.33-8.10 (m, 1H, C4 pyridine proton), 7.66-7.46 (m, 1H, C5 pyridine proton), 7.23-6.70 (m, rH, C6H4), 3.56 (t, 2H, ##STR1451## 2.90 (t, 2H, CH2). Anal. (C14H14N2O2) C, H, N. were gradually added Stirring the formed dicyclohexylurea was removed by filtration The pyridine was removed by distillation in vacuo the residue was triturated with cold water filtered crystallized from 50% aqueous methanol Yield

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To an ice cold suspension of 3.69 g (0.03 mol) nicotinic acid in a solution of 5.2 g (0.03 mol) tyramine hydrochloride in 100 ml of pyridine, 6.18 g (0.03 mol) of dicyclohexylcarbodiimide (DCC) were gradually added while stirring. Stirring was continued at room temperature for 24 hrs and the formed dicyclohexylurea was removed by filtration. The pyridine was removed by distillation in vacuo and the residue was triturated with cold water, filtered and crystallized from 50% aqueous methanol. Yield, 6.25 g (86%), m.p. 179°-181° C. PMR (DMSO-d6 /D2O) δ 9.0-8.66 (m, 2H, C2 and C6 pyridine protons), 8.33-8.10 (m, 1H, C4 pyridine proton), 7.66-7.46 (m, 1H, C5 pyridine proton), 7.23-6.70 (m, rH, C6H4), 3.56 (t, 2H, ##STR1451## 2.90 (t, 2H, CH2). Anal. (C14H14N2O2) C, H, N. were gradually added Stirring the formed dicyclohexylurea was removed by filtration The pyridine was removed by distillation in vacuo the residue was triturated with cold water filtered crystallized from 50% aqueous methanol Yield

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To an ice cold suspension of 3.69 g (0.03 mol) nicotinic acid in a solution of 5.2 g (0.03 mol) tyramine hydrochloride in 100 ml of pyridine, 6.18 g (0.03 mol) of dicyclohexylcarbodiimide (DCC) were gradually added while stirring. Stirring was continued at room temperature for 24 hrs and the formed dicyclohexylurea was removed by filtration. The pyridine was removed by distillation in vacuo and the residue was triturated with cold water, filtered and crystallized from 50% aqueous methanol. Yield, 6.25 g (86%), m.p. 179°-181° C. PMR (DMSO-d6 /D2O) δ 9.0-8.66 (m, H, C2 and C6 pyridine protons), 8.33-8.10 (m, 1H, C4 pyridine proton), 7.66-7.46 (m, 1H, C5 pyridine proton), 7.23-6.70 (m, rH, C6H4), 3.56 ##STR1469## 2.90 (t, 2H, CH2). Anal. (C14H14N2O2) C, H, N. were gradually added Stirring the formed dicyclohexylurea was removed by filtration The pyridine was removed by distillation in vacuo the residue was triturated with cold water filtered crystallized from 50% aqueous methanol Yield

反応条件: temp:96.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: The crude product of 4-pentyloxyphthalic acid was dissolved in acetic acid (20 ml), and tyramine hydrochloride (2.74 g, 20 mmol) was added. The mixture was stirred at 96° C. for 4 hours, and concentrated under reduced pressure. The obtained residue was purified by column chromatography on silica gel (hexane/ethyl acetate=6/1) to give N-2-(4-hydroxyphenyl)ethyl-4-pentyloxyphthalimide (2.6 g, 9.4 mmol, 88%) as colorless crystals. concentrated under reduced pressure The obtained residue was purified by column chromatography on silica gel (hexane/ethyl acetate=6/1)

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To an ice cold suspension of 3.69 g (0.03 mol) nicotinic acid in a solution of 5.2 g (0.03 mol) tyramine hydrochloride in 100 ml of pyridine, 6.18 g (0.03 mol) of dicyclohexylcarbodiimide (DCC) were gradually added while stirring. Stirring was continued at room temperature for 24 hrs and the formed dicyclohexylurea was removed by filtration. The pyridine was removed by distillation in vacuo and the residue was triturated with cold water, filtered and crystallized from 50% aqueous methanol. Yield, 6.25 g (86%), m.p. 179°-181° C. PMR (DMSO-d6 /D2O) δ 9.0-8.66 (m, 2H, C2 and C6 pyridine protons), 8.33-8.10 (m, 1H, C4 pyridine proton), 7.66-7.46 (m, 1H, C5 pyridine proton), 7.23-6.70 (m, rH, C6H4), 3.56 (t, 2H, ##STR1535## 2.90 (t, 2H, CH2). Anal. (C14H14N2O2) C, H, N. were gradually added Stirring the formed dicyclohexylurea was removed by filtration The pyridine was removed by distillation in vacuo the residue was triturated with cold water filtered crystallized from 50% aqueous methanol Yield

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To an ice cold suspension of 3.69 g (0.03 mol) nicotinic acid in a solution of 5.2 g (0.03 mol) tyramine hydrochloride in 100 ml of pyridine, 6.18 g (0.03 mol) of dicyclohexylcarbodiimide (DCC) were gradually added while stirring. Stirring was continued at room temperature for 24 hrs and the formed dicyclohexylurea was removed by filtration. The pyridine was removed by distillation in vacuo and the residue was triturated with cold water, filtered and crystallized from 50% aqueous methanol. Yield, 6.25 g (86%), m.p. 179°-181° C. PMR (DMSO-d6 /D2O) δ 9.0-8.66 (m, 2H, C2 and C6 pyridine protons), 8.33-8.10 (m, 1H, C4 pyridine proton), 7.66-7.46 (m, 1H, C5 pyridine proton), 7.23-6.70 (m, rH, C6H4), 3.56 (t, 2H, ##STR1451## 2.90 (t, 2H, CH2). Anal. (C14H14N2O2) C, H, N. were gradually added Stirring the formed dicyclohexylurea was removed by filtration The pyridine was removed by distillation in vacuo the residue was triturated with cold water filtered crystallized from 50% aqueous methanol Yield

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To a mixture of 1.0 g (3.13 Mol) of Boc-D-Arg(NO2)-OH, 0.57 g (3.28 mMol) of tyramine-hydrochloride, 0.91 ml (0.66 g =6.53 mMol) of triethylamine and 20 ml of anhydrous acetonitrile were added 1.05 g (3.27 mMol) of TBTU, with stirring and external cooling with ice water. The mixture was allowed to come up to ambient temperature and stirring was continued overnight under these conditions. The precipitate was filtered off, the filtrate was evaporated down in vacuo, the residue was distributed between water and ethyl acetate. The ethyl acetate phase was then purified by column chromatography on silica gel (Macherey-Nagel, 35-70 mesh ASTM) using dichloromethane/methanol/cyclohexane/conc. aqueous ammonia =68/15/15/2 (v/v/v/v). 0.8 g (58% of theory) of the title compound were obtained in the form of an amorphous foam, Rf 0.49 (Macherey-Nagel Polygram® SIL G/UV254, ready-made films for TLC; eluant: dichloromethane/methanol/cyclohexane/conc. aqueous ammonia =68/15/15/2 (v/v/v/v)). to come up to ambient temperature stirring The precipitate was filtered off the filtrate was evaporated down in vacuo The ethyl acetate phase was then purified by column chromatography on silica gel (Macherey-Nagel, 35-70 mesh ASTM)

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To a mixture of 1.0 g (3.13 Mol) of Boc-D-Arg(NO2)-OH, 0.57 g (3.28 mMol) of tyramine-hydrochloride, 0.91 ml (0.66 g =6.53 mMol) of triethylamine and 20 ml of anhydrous acetonitrile were added 1.05 g (3.27 mMol) of TBTU, with stirring and external cooling with ice water. The mixture was allowed to come up to ambient temperature and stirring was continued overnight under these conditions. The precipitate was filtered off, the filtrate was evaporated down in vacuo, the residue was distributed between water and ethyl acetate. The ethyl acetate phase was then purified by column chromatography on silica gel (Macherey-Nagel, 35-70 mesh ASTM) using dichloromethane/methanol/cyclohexane/conc. aqueous ammonia =68/15/15/2 (v/v/v/v). 0.8 g (58% of theory) of the title compound were obtained in the form of an amorphous foam, Rf 0.49 (Macherey-Nagel Polygram® SIL G/UV254, ready-made films for TLC; eluant: dichloromethane/methanol/cyclohexane/conc. aqueous ammonia =68/15/15/2 (v/v/v/v)). to come up to ambient temperature stirring The precipitate was filtered off the filtrate was evaporated down in vacuo The ethyl acetate phase was then purified by column chromatography on silica gel (Macherey-Nagel, 35-70 mesh ASTM)

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: To a mixture of 1.0 g (3.13 Mol) of Boc-D-Arg(NO2)-OH, 0.57 g (3.28 mMol) of tyramine-hydrochloride, 0.91 ml (0.66 g=6.53 mMol) of triethylamine and 20 ml of anhydrous acetonitrile were added 1.05 g (3.27 mMol) of TBTU, with stirring and external cooling with ice water. The mixture was allowed to come up to ambient temperature and stirring was continued overnight under these conditions. The precipitate was filtered off, the filtrate was evaporated down in vacuo, the residue was distributed between water and ethyl acetate. The ethyl acetate phase was then purified by column chromatography on silica gel (Macherey-Nagel, 35-70 mesh ASTM) using dichloromethane/methanol/cyclohexane/conc. aqueous ammonia=68/15/15/2 (v/v/v/v). 0.8 g (58% of theory) of the title compound were obtained in the form of an amorphous foam, Rf 0.49 (Macherey-Nagel Polygram® SIL G/UV254, ready-made films for TLC; eluant: dichloromethane/methanol/cyclohexane/conc. aqueous ammonia=68/15/15/2 (v/v/v/v)). to come up to ambient temperature stirring The precipitate was filtered off the filtrate was evaporated down in vacuo The ethyl acetate phase was then purified by column chromatography on silica gel (Macherey-Nagel, 35-70 mesh ASTM)

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: HA (1 g, 2.5 mmol) was dissolved in 100 ml of distilled water, forming an initial solution. Tyramine hydrochloride (162 mg, 0.93 mmol) and 5-aminofluorescein (81 mg, 0.23 mmol in 1.62 ml DMSO) were added to this solution. EDC.HCl (479 mg, 2.5 mmol) and NHS (290 mg, 2.5 mmol) were then added and the pH of the mixture was maintained at 4.7 with 0.1 M NaOH. The solution was stirred overnight at room temperature and then brought to pH 7.0. The solution was next filtered with grade 1 Whatman™ cellulose filter paper to remove unconjugated aminofluorescein that had precipitated. The filtrate was collected into dialysis tubes of molecular cut-off 3500 Da. Then the dialysis and lyophilization procedures described in Example I-A were carried out. The degree of substitution of tyramine was calculated from 1H NMR and the degree of aminofluorescein conjugated was estimated by comparing the absorbance value at 490 nm of 1 mg/ml fluorescence-conjugated HA-Tyr solution to a set of aminofluorescein standards. The degrees of substitution of tyramine and aminofluorescein were 4 and 0.4, respectively. next filtered with grade 1 Whatman™ cellulose filter paper to remove unconjugated aminofluorescein that had precipitated The filtrate was collected into dialysis tubes of molecular cut-off 3500 Da Then the dialysis and lyophilization procedures

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: HA (1 g, 2.5 mmol) was dissolved in 100 ml of distilled water, forming an initial solution. Tyramine hydrochloride (162 mg, 0.93 mmol) and 5-aminofluorescein (81 mg, 0.23 mmol in 1.62 ml DMSO) were added to this solution. EDC.HCl (479 mg, 2.5 mmol) and NHS (290 mg, 2.5 mmol) were then added and the pH of the mixture was maintained at 4.7 with 0.1 M NaOH. The solution was stirred overnight at room temperature and then brought to pH 7.0. The solution was next filtered with grade 1 Whatman™ cellulose filter paper to remove unconjugated aminofluorescein that had precipitated. The filtrate was collected into dialysis tubes of molecular cut-off 3500 Da. Then the dialysis and lyophilization procedures described in Example I-A were carried out. The degree of substitution of tyramine was calculated from 1H NMR and the degree of aminofluorescein conjugated was estimated by comparing the absorbance value at 490 nm of 1 mg/ml fluorescence-conjugated HA-Tyr solution to a set of aminofluorescein standards. The degrees of substitution of tyramine and aminofluorescein were 4 and 0.4, respectively. next filtered with grade 1 Whatman™ cellulose filter paper to remove unconjugated aminofluorescein that had precipitated The filtrate was collected into dialysis tubes of molecular cut-off 3500 Da Then the dialysis and lyophilization procedures

反応条件: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: HA (1 g, 2.5 mmol) was dissolved in 100 ml of distilled water, forming an initial solution. Tyramine hydrochloride (162 mg, 0.93 mmol) and 5-aminofluorescein (81 mg, 0.23 mmol in 1.62 ml DMSO) were added to this solution. EDC.HCl (479 mg, 2.5 mmol) and NHS (290 mg, 2.5 mmol) were then added and the pH of the mixture was maintained at 4.7 with 0.1 M NaOH. The solution was stirred overnight at room temperature and then brought to pH 7.0. The solution was next filtered with grade 1 Whatman™ cellulose filter paper to remove unconjugated aminofluorescein that had precipitated. The filtrate was collected into dialysis tubes of molecular cut-off 3500 Da. Then the dialysis and lyophilization procedures described in Example I-A were carried out. The degree of substitution of tyramine was calculated from 1H NMR and the degree of aminofluorescein conjugated was estimated by comparing the absorbance value at 490 nm of 1 mg/ml fluorescence-conjugated HA-Tyr solution to a set of aminofluorescein standards. The degrees of substitution of tyramine and aminofluorescein were 4 and 0.4, respectively. next filtered with grade 1 Whatman™ cellulose filter paper to remove unconjugated aminofluorescein that had precipitated The filtrate was collected into dialysis tubes of molecular cut-off 3500 Da Then the dialysis and lyophilization procedures