反应详情

温度

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-isopropoxypurine (0.159 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, 0.159 g, 0.8 mmol, of 2-amino-6-isopropoxypurine was added to the reaction. Fifteen days after the reaction was started, another 0.159 g, 0.8 mmol, of 2-amino-6-isopropoxypurine and 0.428 g, 1.6 mmol, of 3'-azido-3'-deoxythymidine were added to the reaction. Twenty-seven days after the reaction was started, 0.159 g, 0.58 mmoles, of 2-amino-6-isopropoxypurine was added to the reaction. The reaction was stopped after thirty-three days. The reaction was heated at 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 was removed in vacuo to give a foam. The foam was chromatographed on basic alumina grade 1 eluted with chloroform/methanol (95:5, v/v). The product containing fractions were combined and the solvent removed in vacuo to give the product as a foam (28% yield). mp=195°-197° C.

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, 0.159 g, 0.8 mmol, of 2-amino-6-isopropoxypurine was added to the reaction
6. customFifteen days
7. additionanother 0.159 g, 0.8 mmol, of 2-amino-6-isopropoxypurine and 0.428 g, 1.6 mmol, of 3'-azido-3'-deoxythymidine were added to the reaction
8. customTwenty-seven days
9. addition0.159 g, 0.58 mmoles, of 2-amino-6-isopropoxypurine was added to the reaction
10. waitThe reaction was stopped after thirty-three days
11. temperatureThe reaction was heated at 80° C.
12. customto precipitate the enzyme
13. filtrationfiltered
14. extractionThe aqueous solution was extracted with ethyl acetate
15. customto remove the product
16. dry with materialThe combined ethyl acetate fractions were dried with magnesium sulfate
17. filtrationfiltered
18. customthe solvent was removed in vacuo
19. customto give a foam
20. customThe foam was chromatographed on basic alumina grade 1
21. washeluted with chloroform/methanol (95:5
22. additionThe product containing fractions
23. customthe solvent removed in vacuo