Reaction #1357216

ord-a70b73ab4ce84406882fac4b0fe6f5d0

Reaction equation

NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
CC#N
acetonitrile
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
[Cl-].[Cl-].[Mg+2]
MgCl2
O=C(O)CN(CCN(CC(=O)O)CC(=O)O)CC(=O)O
EDTA
→
Cc1c[nH]c2ccccc12
Skatole

Reactants

NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
CC#N
acetonitrile
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
[Cl-].[Cl-].[Mg+2]
MgCl2
O=C(O)CN(CCN(CC(=O)O)CC(=O)O)CC(=O)O
EDTA
—
ice

Reagents

None

Solvents

O=P([O-])([O-])[O-].[Na+].[Na+].[Na+]
sodium phosphate

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Otherover a range of 10 to 40 min

Procedure

Microsomal incubations. Two mg microsomal protein was incubated with 0.4 mM 3MI and 4 mM NADPH in 0.05M sodium phosphate buffer (pH 7.4) containing 5 mM MgCl2 and 1 mM EDTA for 30 min at 37.degree. C. (production of metabolites was determined to be linear over a range of 10 to 40 min). Incubation volumes were 0.5 ml. Reactions were started by the addition of NADPH after 3-minute preincubation periods at 37° C., and stopped with 0.5 ml of ice-cold acetonitrile. Incubations of all 30 samples were run in duplicate and for control incubations NADPH was omitted. After the addition of acetonitrile the mixture was vortexed and centrifuged at 5000 rpm for 20 min. A 50 μl aliquot of the clear supernatant was analyzed by high-performance liquid chromatography (HPLC).

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07202035B2uspto-grants-2007_04