Thymidine → 2-amino-6-benzyloxy-9-(2,3-dideoxy-3-fluoro-beta-D-erythro-pentofuranosyl)-9H-purine

HRID 1376445Notes

Reaction equation

HRID 1376445 reaction equation
[NH2:1][C:2]1[N:10]=[C:9]2[C:5]([N:6]=[CH:7][NH:8]2)=[C:4]([O:11][CH2:12][C:13]2[CH:18]=[CH:17][CH:16]=[CH:15][CH:14]=2)[N:3]=1.[F:19][C@@H:20]1[C@@H:24]([CH2:25][OH:26])[O:23][C@@H:22](N2C=CC(=O)NC2=O)[CH2:21]1.[N-]=[N+]=[N-].[K+].[C@@H]1(N2C=C(C)C(=O)NC2=O)O[C@H](CO)[C@@H](O)C1>P([O-])([O-])([O-])=O.[K+].[K+].[K+]>[NH2:1][C:2]1[N:10]=[C:9]2[C:5]([N:6]=[CH:7][N:8]2[C@@H:22]2[O:23][C@H:24]([CH2:25][OH:26])[C@@H:20]([F:19])[CH2:21]2)=[C:4]([O:11][CH2:12][C:13]2[CH:14]=[CH:15][CH:16]=[CH:17][CH:18]=2)[N:3]=1

Participants

6

Reactant 4

HCID 5789Thymidine3×HCID 1624502',3'-Dideoxy-3'-fluorouridineHCID 10290740Potassium azide2×HCID 178508945HCID 178508945, compound

Product 1

HCID 677039612-amino-6-benzyloxy-9-(2,3-dideoxy-3-fluoro-beta-D-erythro-pentofuranosyl)-9H-purineYield 62%

Reagents, catalysts & solvents 1

HCID 62657Potassium Phosphate, TribasicSolvent2×

Conditions

Temperature
37 °C

Procedure

2-Amino-6-benzyloxy-9H-purine (0.60 g 2.5 mmoles) and 2',3'-dideoxy-3'-fluorouridine (0.5 g 2.2 mmoles) were suspended in potassium phosphate buffer (10 mM) 50 ml, pH 6.8, containing 0.04% potassium azide. Purified purine nucleoside phosphorylase (1070) I.U.) and thymidine phosphorylase (400 I.U.) (Krenitsky et al, Biochemistry, 20, 3615 (1981) and U.S. Pat. No. 4,381,344) were added to the reaction mixture and the suspension stirred at 37° C. After 24 hours, additional thymidine phosphorylase (2000 I.U.) was added and, after 14 days, an additional 10 mM potassium phosphate buffer 25 ml, pH 6.8, containing 0.04% potassium azide was added. Additional purine nucleoside phosphorylase (5400 I.U.) and thymidine phosphorylase (2000 I.U.) were added after 17 days. After a total reaction time of 33 days, denatured protein was removed by filtration. The precipitate was washed with methanol until all the UV absorbing material was eluted from the precipitate. The filtrate was diluted with water until the methanol content was <10% and this diluted sample was applied to a series of coupled columns. The initial column contained AG1-X2 (2.5×10 cm, OH form) while the second column contained XAD-2 (2.5×20 cm). After sample application, the columns were washed with water 800 ml and the product eluted with methanol. Product-containing fractions were flash chromatographed on silica gel (2.5×20 cm) using dichloromethane:methanol (9:1). Solvent was removed and lyophilization yielded 2-amino-6-benzyloxy-9-(2,3-dideoxy-3-fluoro-β-D-erythro-pentofuranosyl)-9H-purine (0.49 g).

Workup

  1. addition4,381,344) were added to the reaction mixture
  2. additionwas added
  3. customAfter a total reaction time of 33 days
  4. customwas removed by filtration
  5. washThe precipitate was washed with methanol until all the UV
  6. customabsorbing material
  7. washwas eluted from the precipitate
  8. additionThe filtrate was diluted with water until the methanol content
  9. washAfter sample application, the columns were washed with water 800 ml
  10. washthe product eluted with methanol
  11. customchromatographed on silica gel (2.5×20 cm)
  12. customSolvent was removed

Sources & evidence

ORD record
ord-a17cc8efcc634f85befc4193aefa6ba7
Source dataset
uspto-grants-1997_09
Patent number
US05663154

Reaction notes

Loading…