NADP nicotinamide-adenine-dinucleotide phosphate → HCID 46929323, compound

HRID 1514691Notes

Reaction equation

HRID 1514691 reaction equation
[Cl:1][C:2]1[CH:7]=[CH:6][C:5]([C:8]2[C:12]([CH2:13][O:14][C:15]3[N:20]=[CH:19][C:18]([C:21]([N:23]4[CH2:28][CH2:27][S:26](=[O:30])(=[O:29])[CH2:25][CH2:24]4)=[O:22])=[CH:17][CH:16]=3)=[C:11]([CH3:31])[O:10][N:9]=2)=[CH:4][CH:3]=1.C1C=[N+]([C@@H]2[O:42][C@H](COP(OP(OC[C@H]3O[C@@H](N4C5N=CN=C(N)C=5N=C4)[C@H](OP(O)(O)=O)[C@@H]3O)(O)=O)(O)=O)[C@@H](O)[C@H]2O)C=C(C(N)=O)C=1.[Cl-].[Mg+2].[Cl-].C(#N)C>CS(C)=O>[Cl:1][C:2]1[CH:3]=[CH:4][C:5]([C:8]2[C:12]([CH2:13][O:14][C:15]3[N:20]=[CH:19][C:18]([C:21]([N:23]4[CH2:24][CH2:25][S:26](=[O:30])(=[O:29])[CH2:27][CH2:28]4)=[O:22])=[CH:17][CH:16]=3)=[C:11]([CH2:31][OH:42])[O:10][N:9]=2)=[CH:6][CH:7]=1

Participants

6

Reactant 4

HCID 5886NADP nicotinamide-adenine-dinucleotide phosphate2×HCID 6342AcetonitrileHCID 5360315Magnesium ChlorideHCID 68457576HCID 68457576, compound

Product 1

HCID 46929323HCID 46929323, compoundYield 18%

Reagents, catalysts & solvents 1

HCID 679Dimethyl SulfoxideSolvent

Procedure

A solution of {6-[3-(4-chloro-phenyl)-5-methyl-isoxazol-4-ylmethoxy]-pyridin-3-yl}-(1,1-dioxo-1-thiomorpholin-4-yl)-methanone (44.5 mg, 0.096 mmol) in DMSO (2 mL) was added to a suspension of CYP3A4 (800 nmol)) and NADP in a total volume of 1000 mL reaction buffer (823 mL 0.1M K+ phosphate buffer pH 7.4, 50 mL 1M trisodium citrate, 10 mL 1M magnesium chloride, 2.5 mL 20 mM NADP). The mixture was shaken in a 2 L baffled Erlenmeyer flask at 220 rpm at 27° C. for 4 h, by which time the ratio of substrate:product (12:8 at 3 h, and 11:9 at 4 h) remained almost constant (reaction monitoring: 0.2 mL suspension was treated with 0.2 mL acetonitrile, centrifuged at 13,200 rpm for 2 min, and supernatant was analyzed by HPLC). The suspension (1000 mL) was extracted with 2 L ethyl acetate. The solvent was distilled off and the residue was dissolved in 100 mL water. This was applied to a 10 g reverse phase C18 cartridge which was eluted with a step gradient of acetonitrile in water (15 mL aliquots). Metabolite containing fractions eluting at 35% MeCN were pooled as appropriate and after removal of organic solvent by rotary evaporation were freeze dried to afford the title compound (8.4 mg, 18%) as a white solid. MS: m/e=536.1 [M+CH3COO]−.

Workup

  1. customat 27° C.
  2. customfor 4 h
  3. customproduct (12:8 at 3 h, and 11:9 at 4 h)
  4. extractionThe suspension (1000 mL) was extracted with 2 L ethyl acetate
  5. distillationThe solvent was distilled off
  6. dissolutionthe residue was dissolved in 100 mL water
  7. washwas eluted with a step gradient of acetonitrile in water (15 mL aliquots)
  8. additionMetabolite containing fractions
  9. washeluting at 35% MeCN
  10. customafter removal of organic solvent
  11. customby rotary evaporation
  12. customdried

Sources & evidence

ORD record
ord-8deb158caa514c85869621da8a331882
Source dataset
uspto-grants-2012_07
Patent number
US08222246B2

Reaction notes

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