反应详情

温度

37 °C

2-Amino-6-methoxypurine (2.0 g, 12 mmoles) which may be prepared according to R. W. Balsiger and J. A. Montgomery (J. Org. Chem. 20:1573, 1960) and 1-(2-deoxy-2-fluoro-β-D-ribofuranosyl)uracil (0.88 g; 3.6 mmoles) which may be prepared according to J. F. Codington et al., (J. Org. Chem. 29:558, 1964) were suspended in 250 ml of 5 mM potassium phosphate buffer, pH 7.0, which contained 0.04% (w/v) potassium azide. The pH of the suspension was adjusted to 7.0 with KOH. Thymidine phosphorylase (41,700 I.U.) and purine nucleoside phosphorylase (83.300 I.U.) (T. A. Krenitsky et al., Biochemistry 20:3615, 1981 and U.S. Pat. No. 4,381,344) which had been absorbed onto 10.5 g of DEAE-cellulose (25 ml) were added and the suspension shaken at 37° C. After 24 hours 1.0 g of 2-amino-6-methoxypurine and 250 ml of the above buffer were added and the temperature increased to 50° C. After another 24 hours the reaction was filtered. The filter cake was washed with water and the filtrates combined and the solvent removed under vacuum. The residue was dissolved in warm water and the pH adjusted to pH 9.4 with NH4OH. The solution was applied to a 2.5×15 cm column of anion exchange resin (Bio-Rad AG1X2-hydroxide form). After washing the column with water, the product was eluted with methanol/water (9/1). Product containing fractions were combined and the solvent removed under vacuum. The residue was dissolved in warm water/methanol and filtered through a 0.22 μm-pore nylon membrane filter. Lyophilization of the filtrate yielded 0.91 g of title compound that analyzed as a 0.5 hydrate.

1. custom4,381,344) which had been absorbed onto 10.5 g of DEAE-cellulose (25 ml)
2. additionwere added
3. additionAfter 24 hours 1.0 g of 2-amino-6-methoxypurine and 250 ml of the above buffer were added
4. temperaturethe temperature increased to 50° C
5. filtrationAfter another 24 hours the reaction was filtered
6. washThe filter cake was washed with water
7. customthe solvent removed under vacuum
8. dissolutionThe residue was dissolved in warm water
9. washAfter washing the column with water
10. washthe product was eluted with methanol/water (9/1)
11. additionProduct containing fractions
12. customthe solvent removed under vacuum
13. dissolutionThe residue was dissolved in warm water/methanol
14. filtrationfiltered through a 0.22 μm-pore nylon membrane
15. filtrationfilter