5'-Atp → 5-Hydroxyicosa-6,8,11,14-tetraenoic acid

HRID 963774Notes

Reaction equation

HRID 963774 reaction equation
C(O)[C:2](N)([CH2:5]O)[CH2:3][OH:4].Cl.[Cl-].[Cl-].[Ca+2].P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)[OH:14].C[O:45][C:46]1[CH:47]=[C:48]([CH:55]=[CH:56][C:57](=O)[CH2:58][CH2:59][CH2:60][CH2:61][C:62]2C=C[CH:65]=[CH:64][CH:63]=2)[CH:49]=[C:50](OC)[C:51]=1O>>[CH3:65][CH2:64][CH2:63][CH2:62][CH2:61][CH:60]=[CH:59][CH2:58][CH:57]=[CH:56][CH2:55][CH:48]=[CH:49][CH:50]=[CH:51][CH:46]([OH:45])[CH2:47][CH2:5][CH2:2][C:3]([OH:4])=[O:14]

Participants

5

Reactant 4

HCID 59575'-AtpHCID 93573Tromethamine hydrochlorideHCID 5284359Calcium ChlorideHCID 536807641-(3,5-Dimethoxy-4-hydroxyphenyl)-7-phenyl-1-heptene-3-one

Product 1

HCID 17715-Hydroxyicosa-6,8,11,14-tetraenoic acid

Procedure

76 μl of 50 mM Tris-HCl buffer (pH 7.4) containing 2.56 mM of CaCl2 and 2.56 mM of ATP and 20 μl of 5-lipoxygenase sample prepared above were added to 2μl of a solution containing a prescribed amount of the diarylheptanoide derivative (1) prepared in Example 1 in DSMO. The mixture was incubated at 37° C. After 15 minutes, the reaction was terminated with the addition of 200 μl of a 145:55:0.1 mixture of acetonitrile, methanol, and acetic acid. The reaction mixture was centrifuged (1300 rpm, 30 min., 40° C.). The amount of 5-HETE in the supernatant was measured by HPLC (column: Nova-pak 5C18; eluant: a 60:40 acetonitrile:0.05 M KH2PO4 solution with 5 mM trihexyl bromide added; flow rate: 0.8 ml/min.; wavelength: 235 nm). The amount of 5-HETE produced was calculated from the peak area of the 5-HETE. This procedure was repeated twice to determine the 5-lipoxygenase inhibiting activity of the diarylheptanoide derivative (1) from the average production amount of 5-HETE. The results are shown in FIG. 1, which indicates the 5-lipoxygenase inhibiting activity of the diarylheptanoide derivative (1).

Workup

  1. customprepared above
  2. customwas incubated at 37° C
  3. customthe reaction was terminated with the addition of 200 μl of a 145:55:0.1 mixture of acetonitrile, methanol, and acetic acid
  4. custom(1300 rpm, 30 min., 40° C.)
  5. additioneluant: a 60:40 acetonitrile:0.05 M KH2PO4 solution with 5 mM trihexyl bromide added

Sources & evidence

ORD record
ord-4b90922aaa1a462ab742f0d0c839c61b
Source dataset
uspto-grants-1998_06
Patent number
US05763673

Reaction notes

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