Reaction equation
C1N=C(N)C2N=CN([C@@H:10]3[O:14][C@H:13]([CH2:15]OP(OP(OC[C@H]4O[C@@H](N5C=C(C(N)=O)CC=C5)[C@H](O)[C@@H]4O)(O)=O)(O)=O)[C@@H:12](O)[C@H:11]3OP(O)(O)=O)C=2N=1.[C:49]([O-:52])(=[O:51])C.[NH4+].[NH:54]1C=CC=NC1.[NH:60]1[CH2:68][CH:66]([F:67])[C:64](=[O:65])[NH:63][C:61]1=[O:62].FC(C(O)=O)CN.C1NC(=O)NC(=O)C=1F.[CH3:85][C@H:86]1[O:90][C@@H:89]([N:91]2[C:97](=[O:98])[NH:96][C:94](=O)[C:93]([F:99])=[CH:92]2)[C@H:88]([OH:100])[C@@H:87]1[OH:101]>C(O)C>[CH2:11]1[CH2:10][O:14][CH:13]([N:60]2[C:61](=[O:62])[NH:63][C:64](=[O:65])[C:66]([F:67])=[CH:68]2)[CH2:12]1.[CH3:10][CH2:11][CH2:12][CH2:13][CH2:15][O:52][C:49]([NH:54][C:94]1[C:93]([F:99])=[CH:92][N:91]([C@@H:89]2[O:90][C@H:86]([CH3:85])[C@@H:87]([OH:101])[C@H:88]2[OH:100])[C:97](=[O:98])[N:96]=1)=[O:51]Participants
10
Procedure
100 μM NADPH and 25 μl of cell supernatant (reaction volume; 50 μl) were incubated at 37° C. for 30 min. After the reaction, 5 μl of the reaction solution was subjected to thin layer chromatography (Silica gel 60F254; Merck, Germany) and developed with a mixed solution of ethanol and 1 M ammonium acetate (5:1, v/v). Dihydropyrimidine dehydrogenase activity was determined by measuring the sum of dihydrofluorouracil and 2-fluoro-β-alanine dissociated from [6-14C] 5-FU with a liquid scintillation counter. Drugs used: UFT, TS-1 and capecitabine were synthesized by Taiho Pharmaceutical Co., Ltd. 5′-DFUR was purchased from Roche Japan. [6-14C]-5-FU (1.85 GBq/mmol) was purchased from Moravek Biochemicals, Inc. (Brea, Calif., USA).
Sources & evidence
- ORD record
- ord-08bf9dab220c4455a76fa5eff9def14f
- Source dataset
- uspto-grants-2013_05
- Patent number
- US08440398B2
Linked notes
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