Citrate → HCID 476440, compound

Temperature

50 °C

To 500 ml of an aqueous pH 6.0 50 mM citrate buffer, prepared as described in Example 2c, was added 2-amino-6-propoxypurine (0.0985 g, 0.5 mmol) and 3'-azido-3'-deoxythymidine (0.668 g, 2.5 mmol). Solution was achieved by heating the mixture at 50° C. with sonication. A sample was removed as a control. A 25 mL solution of trans-N-deoxyribosylase (Example 2b) at an activity of 1500 units/mL was added. The reaction was heated at 50° C. Seven days later 0.0985 g, 0.5 mmol, of 2-amino-6-propoxypurine was added to the reaction. Eleven days after the reaction was started, another 0.0985 g, 0.5 mmol, of 2-amino-6-propoxypurine was added to the reaction. The reaction was terminated after twenty-three days. The reaction was heated to 80° C. to precipitate the enzyme and then 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 was removed in vacuo to give a foam. The foam was chromatographed on 180 g of silica gel (230-400 mesh) eluted first with chloroform/methanol (99:1, v/v) then with chloroform/methanol (98:2, v/v). The product containing fractions were combined and the solvent removed in vacuo to give the product as an oil. The oil was dissolved in water and lyophilized to give a solid (56% yield). mp=145°-147° C.

1. customwith sonication
2. customA sample was removed as a control
3. additionA 25 mL solution of trans-N-deoxyribosylase (Example 2b) at an activity of 1500 units/mL was added
4. temperatureThe reaction was heated at 50° C
5. additionSeven days later 0.0985 g, 0.5 mmol, of 2-amino-6-propoxypurine was added to the reaction
6. customEleven days
7. additionanother 0.0985 g, 0.5 mmol, of 2-amino-6-propoxypurine was added to the reaction
8. customThe reaction was terminated after twenty-three days
9. temperatureThe reaction was heated 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 was removed in vacuo
17. customto give a foam
18. customThe foam was chromatographed on 180 g of silica gel (230-400 mesh)
19. washeluted first with chloroform/methanol (99:1, v/v)
20. additionThe product containing fractions
21. customthe solvent removed in vacuo

ORD record

ord-499a2ee96d164fd380b658e67decd5db

Source dataset

uspto-grants-1992_10

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US05153318