1,3-diisopropylurea

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

実験手順: An alternative method for preparing the chemically derivatized solid phase materials of Formula 1 involves reacting the aliphatic anion group of the aminosilane bridge on the surface of the solid phase material with homobifunctional crosslinking reagents containing two amine reactive groups (e.g. succinyl di-N-hydroxybenzotriazole, ethylene glycol bis-(succinimidyl succinate), ethylene glycol bis-(sulfosuccinimidyl succinate), disuccinimidyl tartarate, disulfosuccinimidyl tartarate, disuccinimidyl glutarate, disuccinimidyl suberate). For example, succinyl di-N-hydroxybenzotriazole may be prepared by reacting succinic acid with HBT in the presence of DMF and a water-soluble carbodiimide, such as DICD, at 2 to 5° C. for 2 to 4 hours. A white, insoluble urea by-product, (CH3)2—CH—NH—C(O)—NH—CH—(CH3)2, produced during the reaction, can be separated from the succinyl di-N-hydroxybenzotriazole by centrifuging the mixture and decanting the supernate. Then the decanted succinyl di-N-hydroxybenzotriazole in DMF can be buffered to pH 8.0 to 10.0 with an aqueous buffer and reacted with the aliphatic amino group at the solid phase surface at 20 to 30° C. for 1 to 2 hours. The solid phase material is then removed from the solution, washed with deionized water and Tween 20 and dried to produce the following chemically derivatized fluid-permeable microporous phase material within the definition of Formula 1 as illustrated in the scheme provided immediately below.

反応条件: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: Triethylamine (0.75 mL, 5.5 mmol) was added to a suspension of tetronic acid (500 mg, 5.00 mmol) in dichloromethane (25 mL) to obtain a homogeneous solution. The solution was cooled to 0° C., mixed with 4-N,N-dimethylaminopyridine (200 mg, 1.7 mmol), 3-cyclohexne-1-carboxylic acid (0.65 mL, 5.6 mmol), and then, diisopropylcarbodiimide (1.0 mL, 6.3 mmol), followed by stirring for 30 min. The resulting mixture was removed from the ice bath, stirred for 14 hours at room temperature, and filtered to remove thus-produced insoluble diisopropyl urea; the reaction solution was washed twice with 5% hydrochloric acid (20 mL) and the water layer was extracted twice with dichloromethane (10 mL). The organic layers were combined, dried over sodium sulfate anhydride, and concentrated; the thus-obtained residue was purified by silica gel column chromatography (silica gel, ethyl acetate:methanol=15:1). The resultant was arranged to be a sodium salt by adding an excess amount of sodium hydrogencarbonate and distilled water (100 mL), and then washed with dichloromethane; the water layer was acidified by adding 5% hydrochloric acid and extracted four times with dichloromethane (20 mL). The resultant was dried over sodium sulfate anhydride and concentrated to give pale yellow crystals (394 mg, 38%). to obtain a homogeneous solution The resulting mixture was removed from the ice bath stirred for 14 hours at room temperature filtered to remove

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

実験手順: Then 11.5 g (50 mM) of Phenylalanine Ethyl Ester hydrochloride and 7.75 mL (45 mM) of diisopropylcarbodiimide were added. Then 8.6 mL (50 mM) of diisopropylethylamine was added via the dropping funnel for 10 min. The reaction mixture was kept for 20 hours at a room temperature, then 10 mL of H2O was added, a precipitate of diisopropylurea formed was filtered off, a resulted solution was analyzed by HPLC. According analysis, the reaction was completed at 50%.

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

実験手順: Then 11.5 g (50 mM) of Phenylalanine Ethyl Ester hydrochloride and 7.75 mL (45 mM) of diisopropylcarbodiimide were added. Then 8.6 mL (50 mM) of diisopropylethylamine was added via the dropping funnel for 10 min. The reaction mixture was kept for 20 hours at a room temperature, then 10 mL of H2O was added, a precipitate of diisopropylurea formed was filtered off, a resulted solution was analyzed by HPLC. According analysis, the reaction was completed at 50%.

反応条件: temp:90.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: 1,3-Diisopropyl-2,4,6-(1H,3H,5H)-pyrimidinetrione was prepared by the dropwise addition of acetic anhydride (40 ml) over three hours to a solution of 1,3-diisopropylurea (7.5 g, 0.05 mole) and malonic acid (5.2 g, 0.05 mole) at 65° C. After the addition was completed, the temperature was raised to 90° C. and maintained for three hours. The solvent was evaporated under reduced pressure and the residue solidified upon cooling. 1,3-Diisopropyl-2,4,6-(1H,3H,5H)-pyrimidinetrione recrystallized from cyclohexane as colorless needles (7.3 g, 70% yield), m.p. 79°-82°. After the addition maintained for three hours The solvent was evaporated under reduced pressure upon cooling

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

実験手順: An alternative method for preparing the chemically derivatized solid phase materials of Formula 1 involves reacting the aliphatic anion group of the aminosilane bridge on the surface of the solid phase material with homobifunctional crosslinking reagents containing two amine reactive groups (e.g. succinyl di-N-hydroxybenzotriazole, ethylene glycol bis-(succinimidyl succinate), ethylene glycol bis-(sulfosuccinimidyl succinate), disuccinimidyl tartarate, disulfosuccinimidyl tartarate, disuccinimidyl glutarate, disuccinimidyl suberate). For example, succinyl di-N-hydroxybenzotriazole may be prepared by reacting succinic acid with HBT in the presence of DMF and a water-soluble carbodiimide, such as DICD, at 2 to 5° C. for 2 to 4 hours. A white, insoluble urea by-product, (CH3)2—CH—NH—C(O)—NH—CH—(CH3)2, produced during the reaction, can be separated from the succinyl di-N-hydroxybenzotriazole by centrifuging the mixture and decanting the supernate. Then the decanted succinyl di-N-hydroxybenzotriazole in DMF can be buffered to pH 8.0 to 10.0 with an aqueous buffer and reacted with the aliphatic amino group at the solid phase surface at 20 to 30° C. for 1 to 2 hours. The solid phase material is then removed from the solution, washed with deionized water and Tween 20 and dried to produce the following chemically derivatized fluid-permeable microporous phase material within the definition of Formula 1 as illustrated in the scheme provided immediately below.

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

実験手順: An alternative method for preparing the chemically derivatized solid phase materials of Formula 1 involves reacting the aliphatic anion group of the aminosilane bridge on the surface of the solid phase material with homobifunctional crosslinking reagents containing two amine reactive groups (e.g. succinyl di-N-hydroxybenzotriazole, ethylene glycol bis-(succinimidyl succinate), ethylene glycol bis-(sulfosuccinimidyl succinate), disuccinimidyl tartarate, disulfosuccinimidyl tartarate, disuccinimidyl glutarate, disuccinimidyl suberate). For example, succinyl di-N-hydroxybenzotriazole may be prepared by reacting succinic acid with HBT in the presence of DMF and a water-soluble carbodiimide, such as DICD, at 2 to 5° C. for 2 to 4 hours. A white, insoluble urea by-product, (CH3)2—CH—NH—C(O)—NH—CH—(CH3)2, produced during the reaction, can be separated from the succinyl di-N-hydroxybenzotriazole by centrifuging the mixture and decanting the supernate. Then the decanted succinyl di-N-hydroxybenzotriazole in DMF can be buffered to pH 8.0 to 10.0 with an aqueous buffer and reacted with the aliphatic amino group at the solid phase surface at 20 to 30° C. for 1 to 2 hours. The solid phase material is then removed from the solution, washed with deionized water and Tween 20 and dried to produce the following chemically derivatized fluid-permeable microporous phase material within the definition of Formula 1 as illustrated in the scheme provided immediately below.

反応条件: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: Triethylamine (0.75 mL, 5.5 mmol) was added to a suspension of tetronic acid (500 mg, 5.00 mmol) in dichloromethane (25 mL) to obtain a homogeneous solution. The solution was cooled to 0° C., mixed with 4-N,N-dimethylaminopyridine (200 mg, 1.7 mmol), 3-cyclohexne-1-carboxylic acid (0.65 mL, 5.6 mmol), and then, diisopropylcarbodiimide (1.0 mL, 6.3 mmol), followed by stirring for 30 min. The resulting mixture was removed from the ice bath, stirred for 14 hours at room temperature, and filtered to remove thus-produced insoluble diisopropyl urea; the reaction solution was washed twice with 5% hydrochloric acid (20 mL) and the water layer was extracted twice with dichloromethane (10 mL). The organic layers were combined, dried over sodium sulfate anhydride, and concentrated; the thus-obtained residue was purified by silica gel column chromatography (silica gel, ethyl acetate:methanol=15:1). The resultant was arranged to be a sodium salt by adding an excess amount of sodium hydrogencarbonate and distilled water (100 mL), and then washed with dichloromethane; the water layer was acidified by adding 5% hydrochloric acid and extracted four times with dichloromethane (20 mL). The resultant was dried over sodium sulfate anhydride and concentrated to give pale yellow crystals (394 mg, 38%). to obtain a homogeneous solution The resulting mixture was removed from the ice bath stirred for 14 hours at room temperature filtered to remove

反応条件: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

実験手順: Triethylamine (0.75 mL, 5.5 mmol) was added to a suspension of tetronic acid (500 mg, 5.00 mmol) in dichloromethane (25 mL) to obtain a homogeneous solution. The solution was cooled to 0° C., mixed with 4-N,N-dimethylaminopyridine (200 mg, 1.7 mmol), 3-cyclohexne-1-carboxylic acid (0.65 mL, 5.6 mmol), and then, diisopropylcarbodiimide (1.0 mL, 6.3 mmol), followed by stirring for 30 min. The resulting mixture was removed from the ice bath, stirred for 14 hours at room temperature, and filtered to remove thus-produced insoluble diisopropyl urea; the reaction solution was washed twice with 5% hydrochloric acid (20 mL) and the water layer was extracted twice with dichloromethane (10 mL). The organic layers were combined, dried over sodium sulfate anhydride, and concentrated; the thus-obtained residue was purified by silica gel column chromatography (silica gel, ethyl acetate:methanol=15:1). The resultant was arranged to be a sodium salt by adding an excess amount of sodium hydrogencarbonate and distilled water (100 mL), and then washed with dichloromethane; the water layer was acidified by adding 5% hydrochloric acid and extracted four times with dichloromethane (20 mL). The resultant was dried over sodium sulfate anhydride and concentrated to give pale yellow crystals (394 mg, 38%). to obtain a homogeneous solution The resulting mixture was removed from the ice bath stirred for 14 hours at room temperature filtered to remove

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

実験手順: N-Hydroxysuccinimide (1.70 g, 14.77 mmol) and DIC (1.87 g, 14.77 mmol) were added sequentially to a stirred solution of Fmoc-Gly-OH (4.0 g, 13.45 mmol) in dichloromethane (15 mL); the resulting mixture was stirred at room temperature for 4 h. The N,N′-diisopropylurea formed was removed by filtration and the solid was washed with ether (20 mL). The volatiles were removed and the solid Fmoc-Gly-succinimidyl ester formed was washed with ether (3×20 mL). Fmoc-Gly-succinimidyl ester was then redissolved in dry DMF (15 mL) and 3-aminodeoxycholic acid (5.21 g, 12.78 mmol) was added to the clear solution. The reaction mixture was stirred at room temperature for 4 h, water (200 mL) was added and the precipitated solid was filtered, washed with water, dried and purified by silica gel chromatography (TLC (silica): (Rf: 0.50, silica gel, CH2Cl2/CH3OH, 9:1) (eluant: CH2Cl2/CH3OH (9:1)) to give (3β,5β,7α,12α)-3-[[(9H-Fluoren-9-ylmethoxy)amino]acetyl]amino-7,12-dihydroxycholan-24-oic acid as a colorless solid. Yield: 7.46 g (85%).

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

実験手順: N-Hydroxysuccinimide (1.70 g, 14.77 mmol) and DIC (1.87 g, 14.77 mmol) were added sequentially to a stirred solution of Fmoc-Gly-OH (4.0 g, 13.45 mmol) in dichloromethane (15 mL); the resulting mixture was stirred at room temperature for 4 h. The N,N′-diisopropylurea formed was removed by filtration and the solid was washed with ether (20 mL). The volatiles were removed and the solid Fmoc-Gly-succinimidyl ester formed was washed with ether (3×20 mL). Fmoc-Gly-succinimidyl ester was then redissolved in dry DMF (15 mL) and 3-aminodeoxycholic acid (5.21 g, 12.78 mmol) was added to the clear solution. The reaction mixture was stirred at room temperature for 4 h, water (200 mL) was added and the precipitated solid was filtered, washed with water, dried and purified by silica gel chromatography (TLC (silica): (Rf: 0.50, silica gel, CH2Cl2/CH3OH, 9:1) (eluant: CH2Cl2/CH3OH (9:1)) to give (3β,5β,7α,12α)-3-[[(9H-Fluoren-9-ylmethoxy)amino]acetyl]amino-7,12-dihydroxycholan-24-oic acid as a colorless solid. Yield: 7.46 g (85%).

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

実験手順: N-Hydroxysuccinimide (1.70 g, 14.77 mmol) and DIC (1.87 g, 14.77 mmol) were added sequentially to a stirred solution of Fmoc-Gly-OH (4.0 g, 13.45 mmol) in dichloromethane (15 mL); the resulting mixture was stirred at room temperature for 4 h. The N,N′-diisopropylurea formed was removed by filtration and the solid was washed with ether (20 mL). The volatiles were removed and the solid Fmoc-Gly-succinimidyl ester formed was washed with ether (3×20 mL). Fmoc-Gly-succinimidyl ester was then redissolved in dry DMF (15 mL) and 3-aminodeoxycholic acid (5.21 g, 12.78 mmol) was added to the clear solution. The reaction mixture was stirred at room temperature for 4 h, water (200 mL) was added and the precipitated solid was filtered, washed with water, dried and purified by silica gel chromatography (TLC (silica): (Rf: 0.50, silica gel, CH2Cl2/CH3OH, 9:1) (eluant: CH2Cl2/CH3OH (9:1)) to give (3β,5β,7α,12α)-3-[[(9H-Fluoren-9-ylmethoxy)amino]acetyl]amino-7,12-dihydroxycholan-24-oic acid as a colorless solid. Yield: 7.46 g (85%).