Thymidine → HCID 9861413, compound

Participants

Reaction Conditions

Reaction Steps

2-Amino-6-(cyclopropylamino)-9H-purine was prepared by the displacement of the clorine group on 2-amino-6-chloropurine (Aldrich Chemical Company) by cyclopropylamine (Aldrich Chemical Company) 2-Amino-6-(cyclopropylamino)-9H-purine (0.50 g, 2.7 mmoles) was dissolved in DMF (5 ml) and DMSO (5 ml). 3'-Deoxy-3'-fluorothymidine (0.81 g, 3.3 mmoles) and potassium phosphate buffer (10 mM, 30 ml), pH 6.8, containing 0.04% potassium azide were added. Purified purine nucleoside phosphorylase (7500 I.U.) and thymidine phosphorylase (37.50 I.U.) adsorbed onto 5 ml DEAE-cellulose resin were added to the reaction and the suspension stirred at 37° C. After 23 days, the reaction was filtered and the filtrate applied to a series of coupled columns. The initial column contained AG1-X2 (OH-form, 2.5×10 cm) while the second column contained Amberlite XAD-2 resin (2.5×20 cm). After sample application, the columns were washed with water (700 ml) and the product eluted with methanol. Product-containing fractions were then flash chromatographed down a silica gel column (4.8×24 cm) using ethyl acetate:methanol (95:5) as eluant. The product was further purified by flash chromatography down a silica gel column (2.5×40 cm) using dichloromethane:methanol(9:1) as eluant. Solvent was evaporated in vacuo and lyophilisation yielded 0.30 g of 2-amino-6-(cyclopropyl-amino)-9-(2,3-dideoxy-3-fluoro-β-D-erythro-pentofuranosyl)-9H-purine (33%): mp 75° C.;

Provenance & Evidence