3-aminobenzene-1,2-diol

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

実験手順: To a solution of 3-nitrobenzene-1,2-diol (600 mg) in methanol (6 ml) was added 10% palladium on carbon (60 mg), and the mixture was stirred for 3 hours at ambient temperature under hydrogen atmosphere. Insoluble material was filtered off, and the filtrate was concentrated in vacuo. The residue was crystallized with diisopropyl ether to give 3-aminobenzene-1,2,-diol (470 mg). Insoluble material was filtered off the filtrate was concentrated in vacuo The residue was crystallized with diisopropyl ether

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

実験手順: DOPA decarboxylase activity was routinely measured by liquid chromatography and electrochemical detection of catecholamine products. DOPA decarboxylase (1-5 micrograms, 1-20 units) was incubated with L-DOPA (2000 micromolar, 150 microliters) and PLP (10 micromolar) in 0.1M phosphate at pH 6.8 and 37° C. After 1-10 minutes, the reaction was quenched with citric acid (2M, 25 microliters), diluted two-fold with distilled water and 10 microliter aliquots were analyzed by HPLC on a reverse phase Brownlee RP-18 column. Isocratic elution with pH 4.35 buffer [0.1M NaOAc (63% v/v), 0.1M citric acid (32% v/v) and methanol (15% v/v)] at a flow rate of 2.5 ml/min. afforded baseline resolution of dopamine (RT=2.8 min.) from L-DOPA (RT=1.9 min.). the reaction was quenched with citric acid (2M, 25 microliters) diluted two-fold with distilled water and 10 microliter aliquots Isocratic elution with pH 4.35 buffer [0.1M NaOAc (63% v/v), 0.1M citric acid (32% v/v) and methanol (15% v/v)] at a flow rate of 2.5 ml/min.

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

実験手順: To a solution of 3-nitrobenzene-1,2-diol (600 mg) in methanol (6 ml) was added 10% palladium on carbon (60 mg), and the mixture was stirred for 3 hours at ambient temperature under hydrogen atmosphere. Insoluble material was filtered off, and the filtrate was concentrated in vacuo. The residue was crystallized with diisopropyl ether to give 3-aminobenzene-1,2,-diol (470 mg). Insoluble material was filtered off the filtrate was concentrated in vacuo The residue was crystallized with diisopropyl ether

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

実験手順: Chromaffin cells were prepared from bovine adrenal glands by protease perfusion using the method described by Livett in Physiol. Rev. 64:1103 (1984). The cells were plated at 1×106 cells/well in 24-well plates in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum, 8 μM fluorodoxyurine, 50 μg/ml gentamicin, 10 μM cytosine arabinofuranoside, 2.5 μg/ml fungizone, 25 international units/ml penicillin, 25 μg/ml streptomycin and 2 mM glutamine. Experiments were performed 3-8 days after plating. Ca2+-evoked catecholamine release was measured fluorometrically. Chromaffin cells were prepared from bovine adrenal glands by protease perfusion

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

実験手順: The whole brains of mice of ddY-strain were homogenized and the resulting brain homogenate was diluted with the same volume of a buffere solution (comprising 50 mM Tris-HCl (pH 8.6, 1.5 mM EDTA.2Na); 0.5 mM Pridoxal-5-Phosphate; 1 mM Nialamide; and 0.85 mM L-ascorbic acid), and the mixture was centrifuged at 10,000 g to give a supernatant liquid containing the brain enzymes. To this supernatant liquid (500 μl) was added a solution containing 4.5 mM of a substrate selected from L-threo-adrenalinecarboxylic acid, L-threo-DOPS and L-DOPA. The mixture obtained (pH ca. 8, 1.0 ml) was incubated for 30 minutes at 37° C., followed by quantitative determination of adrenaline, noradrenaline or dopamine formed as the reaction product catecholamine. to give a supernatant liquid containing the brain enzymes containing 4.5 mM of The mixture obtained (pH ca. 8, 1.0 ml)

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

実験手順: A mixture of 2,3-dihydroxyaniline (2.5 g.), triethylorthoformate (4.5 g.) and 0.1 g. sulfuric acid is heated at 130° C. for one hour. After cooling, the remaining material is purified by HPLC on silica gel. After cooling the remaining material is purified by HPLC on silica gel

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

実験手順: To a solution of 3-nitrobenzene-1,2-diol (600 mg) in methanol (6 ml) was added 10% palladium on carbon (60 mg), and the mixture was stirred for 3 hours at ambient temperature under hydrogen atmosphere. Insoluble material was filtered off, and the filtrate was concentrated in vacuo. The residue was crystallized with diisopropyl ether to give 3-aminobenzene-1,2,-diol (470 mg). Insoluble material was filtered off the filtrate was concentrated in vacuo The residue was crystallized with diisopropyl ether

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

実験手順: DOPA decarboxylase activity was routinely measured by liquid chromatography and electrochemical detection of catecholamine products. DOPA decarboxylase (1-5 micrograms, 1-20 units) was incubated with L-DOPA (2000 micromolar, 150 microliters) and PLP (10 micromolar) in 0.1M phosphate at pH 6.8 and 37° C. After 1-10 minutes, the reaction was quenched with citric acid (2M, 25 microliters), diluted two-fold with distilled water and 10 microliter aliquots were analyzed by HPLC on a reverse phase Brownlee RP-18 column. Isocratic elution with pH 4.35 buffer [0.1M NaOAc (63% v/v), 0.1M citric acid (32% v/v) and methanol (15% v/v)] at a flow rate of 2.5 ml/min. afforded baseline resolution of dopamine (RT=2.8 min.) from L-DOPA (RT=1.9 min.). the reaction was quenched with citric acid (2M, 25 microliters) diluted two-fold with distilled water and 10 microliter aliquots Isocratic elution with pH 4.35 buffer [0.1M NaOAc (63% v/v), 0.1M citric acid (32% v/v) and methanol (15% v/v)] at a flow rate of 2.5 ml/min.