NADPH → HCID 68182150, compound

HRID 1498042Notes

Reaction equation

HRID 1498042 reaction equation
[CH3:1][NH:2][C:3]1[C:8]2=[C:9]([C:13]3[CH:14]=[N:15][N:16]([CH3:28])[C:17]=3[C:18]3[CH:23]=[CH:22][C:21]([C:24]([F:27])([F:26])[F:25])=[CH:20][CH:19]=3)[N:10]=[C:11]([CH3:12])[N:7]2[N:6]=[CH:5][N:4]=1.C1N=C(N)C2N=CN([C@@H]3[O:42][C@H](COP(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](O)[C@H]3OP(O)(O)=O)C=2N=1>>[CH3:1][NH:2][C:3]1[C:8]2=[C:9]([C:13]3[CH:14]=[N:15][N:16]([CH3:28])[C:17]=3[C:18]3[CH:19]=[CH:20][C:21]([C:24]([F:26])([F:27])[F:25])=[CH:22][CH:23]=3)[N:10]=[C:11]([CH2:12][OH:42])[N:7]2[N:6]=[CH:5][N:4]=1

Participants

3

Reactant 2

HCID 5884NADPHHCID 60163378Imidazo[5,1-f][1,2,4]triazin-4-amine, N,7-dimethyl-5-[1-methyl-5-[4-(trifluoromethyl)phenyl]-1H-pyrazol-4-yl]-

Product 1

HCID 68182150HCID 68182150, compound

Procedure

N,7-Dimethyl-5-{1-methyl-5-[4-(trifluoromethyl)phenyl]-1H-pyrazol-4-yl}imidazo[5,1-f][1,2,4]triazin-4-amine, at a substrate concentration of 50 μM, was incubated for 1 hour with rat liver microsomes and NADPH (1 mM) (21.1 mg/mL protein concentration; 25 mL incubation volume). Incubations were extracted with 4 volumes of acetonitrile. Following centrifugation at 3400 rpm, the supernatant was evaporated at 25° C. in a Turbovap. The residue was reconstituted with 5% aqueous acetonitrile and subjected to preparative reverse-phase HPLC purification (Column: Zorbax Rx-C8 [Agilent], 250×9.6 mm, 5 μm; Mobile phase A: aqueous 5 mM ammonium formate, pH 3; Mobile phase B: acetonitrile; Gradient: 10% to 90% B; UV detection at 254 mm). The fractions containing the M+16 metabolite were dried down to provide the product as a solid. MS m/z 404 (M+H). 1H NMR (600 MHz, DMSO-d6) δ 2.71 (d, J=4.8 Hz, 3H), 3.86 (s, 3H), 4.69 (s, 2H), 6.68 (br q, J=4.7 Hz, 1H), 7.55 (d, J=8.0 Hz, 2H), 7.70 (s, 1H), 7.76 (d, J=8.0 Hz, 2H), 7.85 (s, 1H).

Workup

  1. extractionIncubations were extracted with 4 volumes of acetonitrile
  2. customthe supernatant was evaporated at 25° C. in a Turbovap
  3. customsubjected to preparative reverse-phase HPLC purification (Column: Zorbax Rx-C8 [Agilent], 250×9.6 mm, 5 μm; Mobile phase A: aqueous 5 mM ammonium formate, pH 3; Mobile phase B: acetonitrile; Gradient: 10% to 90% B; UV detection at 254 mm)
  4. additionThe fractions containing the M+16 metabolite
  5. customwere dried down

Sources & evidence

ORD record
ord-d89ee7736591469b8b40f108771b829d
Source dataset
uspto-grants-2015_12
Patent number
US09200000B2

Reaction notes

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