1,3-dicyclohexylurea

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of dicyclohexylcarbodiimide (0.084 g, 0.216 mmol) in dichloromethane (1 ml) was added to a solution of levulinic acid (0.047 g, 0.406 mmol) in dichloromethane (2 ml). A white crystalline solid of dicyclohexylurea formed immediately. The suspension was allowed to stand at room temperature for 30 min, then chilled in an ice bath for 3 h min, and left to stand at 4° overnight before filtration. The colourless filtrate was concentrated to give a colourless oil with a small amount of suspended white solid of dicyclohexylcarbodiimide (DCC) and dicyclohexylurea (DCU). This was used as crude in the preparation of (3).

Conditions: temp:70.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Formation of the tyramide of compound III, is as follows. N-hydroxysuccinimide (33 mg) is dissolved in the reaction mixture prepared above by heating to 70° C., then dicyclohexylcarbodiimide (61 mg) is added. This is reacted at 70° C. for 90 minutes, an additional amount (6 mg) of the carbodiimide is added and the reaction is continued for another 30 minutes. The mixture is cooled to 0° C. and the precipitated dicyclohexylurea is settled by centrifugation. The supernatant, containing 6-(-4′-N-hydroxylsuccinimidyl carboxy phenyl)-3,8-bis bromoacetamidyl-5-methylphenanthridinium chloride (IV) is recovered. A solution of tyramine [4-(2′-amino)ethylphenol] (1.0 g) in water (5 ml) is formed by bringing it to pH 7 by addition of concentrated HCl. This solution is added to the supernatant containing compound IV and reacted at room temperature for one hour. This mixture is then reduced to a red oil by evacuation at 60° C., and is then extracted with water to remove excess tyramine. Compound (VI) is then recrystalized from ethanol-acidified with HCl. prepared above This is reacted at 70° C. for 90 minutes The mixture is cooled to 0° C. is recovered

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Into 100 ml of dimethylformamide, 23.8 g of Compound (A), 30.1 g of Compound (B) and 0.8 g of 4-dimethylamino-pyridine were dissolved, and thereto an ethyl acetate 20 ml solution containing 20.6 g of dicyclohexylcarbodiimide was added dropwise in an ice bath. After the dropwise addition, the solution was allowed to react at room temperature over night. Into the reaction solution, 200 ml of ethyl acetate was poured, and dicyclohexylurea obtained was separated by filtration. Into the filtrate, 300 ml of water was poured to perform liquid separation operation. Water washing was performed twice, the organic layer was dried over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the oily product obtained was subjected to silica gel column chromatography. As a result, from the methylene chloride/ethyl acetate=10/3 fraction, 43 g of Compound (C) was obtained as a pale yellow oily product. ##STR16## Synthesis of Compound (D): was added dropwise in an ice bath After the dropwise addition to react at room temperature over night

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 3.16 g of tetrahydro-3-furoic acid (27 mmol, 1 equiv.) in 25 mL anhydrous dichloromethane was added 7.85 g of dicyclohexylcarbodiimide (38 mmol, 1.4 equiv.) and 4.54 mL triethylamine (32.6 mmol, 1.2 equiv.). N-methyl-methanolamine hydrochloride was then added, followed by 60 mg of DMAP (4-dimethylamino)pyridine. An exothermic reaction ensued and a further 25 mL of dichloromethane were added. The mixture was stirred at ambient temperature overnight, then filtered through diatomaceous earth and concentrated. The residue was treated with ether and the white solid filtered off and removed. The solvent was removed from the mother liquor and the residue purified by column chromatography on SiO2 using 15-25% EtOAc in hexanes as eluent mixtures to provide the desired amide as a colorless oil (55% of theoretical yield) that still contained 10% of dicyclohexyl urea. This was used without further purification in the next reaction. An exothermic reaction filtered through diatomaceous earth concentrated The residue was treated with ether the white solid filtered off removed The solvent was removed from the mother liquor the residue purified by column chromatography on SiO2 using 15-25% EtOAc in hexanes as eluent mixtures to provide

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 1.65 g (8.98 mmol) of 4-aminodiphenylamine, 1.77 g (8.98 mmol) of intermediate 3.2 and 1.27 g (9.42 mmol) of hydroxybenzotriazole are dissolved in 40 ml of THF in a 100 ml flask, under a nitrogen atmosphere. When all have dissolved, 1.94 g (9.42 mmol) of 1,3-dicyclohexylcarbodiimide is added and the reaction medium is left under agitation for 15 hours. The precipitate of dicyclohexylurea formed is filtered and rinsed with ethyl acetate. The filtrate is evaporated under vacuum and the evaporation residue is taken up in ethyl acetate, it then forms a precipitate which is filtered and rinsed using the same solvent. A greenish solid is obtained with a yield of 65%. The product obtained is sufficiently pure to be used directly in the following stage. Melting point: 192-195° C. When all have dissolved

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: DCC (1.0 g, 4.84 mmol) was added to a stirred solution of the product of Example 13d (0.9 g, 4.40 mmol), 2-{1-[(4-chlorophenyl)carbonyl]-5-methoxy-2-methylindol-3-yl}acetic acid (1.57 g, 4.40 mmol), and DMAP (0.05 g) in CH2Cl2 (30 mL) at room temperature. The resulting suspension was stirred for 1 hour at room temperature. The precipitate was removed by filtration and washed with CH2Cl2 (25 mL). The filtrate was concentrated to give a green oil which was chromatographed on silica gel eluting with 5:95 MeOH:CH2Cl2 to afford a green solid (contaminated with dicyclohexyl urea). The solid was triturated with hexane (50 mL) and filtered. The filtrate was concentrated to give the title compound (1.85 g, 77%) as a viscous green oil. 1H NMR (300 MHz, CDCl3) δ 7.47-7.65 (mult, 4H), 6.84-6.91 (mult, 2H), 6.65 (dd, J=2.5 and 9.0 Hz, 1H), 4.29 (t, J=6.7 Hz, 2H), 3.81 (s, 3H), 3.61 (s, 2H), 2.35 (s, 3H), 2.27 (s, 3H), 2.10-2.70 (mult, 10H); 13C NMR (75 MHz, CDCl3) δ 170.5, 168.2, 156.1, 139.2, 135.9, 133.9, 131.1, 130.7, 130.6, 129.1, 114.9, 112.2, 111.6, 101.3, 61.3, 57.5, 55.7, 51.4 (2C), 46.1, 36.6, 30.3, 13.3; mass spectrum (API-TIS) m/z 544 (M+H). The precipitate was removed by filtration washed with CH2Cl2 (25 mL) The filtrate was concentrated to give a green oil which was chromatographed on silica gel eluting with 5:95 MeOH