Citrate → HCID 476443, compound

Temperature

50 °C

To 800 mL of an aqueous pH 6.0, 50 mm citrate buffer, prepared as described in Example 2c' was added 2-amino-6-phenoxy-9H-purine (0.181 g, 0.8 mmol) and 3'-azido-3'-deoxythymidine (1.069 g, 4.0 mmol). Solution was achieved by heating the mixture at 50° C. with sonication. A sample was removed as a control. A 40 mL solution of trans-N-deoxyribosylase (Example 2b) with an activity of 1500 units/mL was added. The reaction was heated at 50° C. Six days later, 0.181 g, 0.8 mmol, of 2-amino-6-phenoxypurine was added to the reaction. Fifteen days after the reaction was started, another 0.181 g, 0.8 mmol, of 2-amino-6-phenoxypurine and 0.428 g, 1.6 mmol, of 3'-azido-3'-deoxythymidine were added to the reaction. The reaction was stopped after thirty-three days, by heating to 80° C. to precipitate the enzyme, and filtered. The aqueous solution was extracted with ethyl acetate to remove the product. The combined ethyl acetate fractions were dried with magnesium sulfate, filtered and the solvent removed in vacuo to give a foam. The foam was chromatographed on basic alumina grade 1 eluted with chloroform/methanol (99:1, v/v). The fractions containing product were combined and the solvent removed in vacuo to give the product as a foam, 0.48 g, 54% yield.

1. customwith sonication
2. customA sample was removed as a control
3. additionA 40 mL solution of trans-N-deoxyribosylase (Example 2b) with an activity of 1500 units/mL was added
4. temperatureThe reaction was heated at 50° C
5. additionSix days later, 0.181 g, 0.8 mmol, of 2-amino-6-phenoxypurine was added to the reaction
6. customFifteen days
7. additionanother 0.181 g, 0.8 mmol, of 2-amino-6-phenoxypurine and 0.428 g, 1.6 mmol, of 3'-azido-3'-deoxythymidine were added to the reaction
8. waitThe reaction was stopped after thirty-three days
9. temperatureby heating to 80° C.
10. customto precipitate the enzyme
11. filtrationfiltered
12. extractionThe aqueous solution was extracted with ethyl acetate
13. customto remove the product
14. dry with materialThe combined ethyl acetate fractions were dried with magnesium sulfate
15. filtrationfiltered
16. customthe solvent removed in vacuo
17. customto give a foam
18. customThe foam was chromatographed on basic alumina grade 1
19. washeluted with chloroform/methanol (99:1, v/v)
20. additionThe fractions containing product
21. customthe solvent removed in vacuo

ORD record

ord-e1b0d82dd8244da0b57b5a3e11e0d047

Source dataset

uspto-grants-1992_10

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05153318